Effects of Diabetes on a Male

基因组学与应用生物学，2014年，第33卷，第5期，第970-974页Genomics and Applied Biology,2014,Vol.33,No.5,970-974研究报告Research Report姜黄素通过调节肠道菌群可改善脂多糖诱导糖尿病刘硒碲1夏宁2*梁瑜祯11广西医科大学第一附属医院,南宁,530021;2广西壮族自治区卫生厅,南宁,530021*通讯作者,xianing12@摘要姜黄素是从姜科植物姜黄的根茎中提取的多酚类物质，具有重要的药用价值。

本研究将30只健康雄性Wistar大鼠，随机分为3组：正常对照组(n=10)、糖尿病模型组(n=10)、糖尿病模型+姜黄素治疗组(n=10简称姜黄素治疗组)。

以LPS(300μg·kg·-1day·-1)皮下注射8周建立2型糖尿病模型。

测定空腹血糖≥11.1mmol/L诊断为糖尿病。

造模成功后，予姜黄素200mg/(kg·-1day·-1)灌胃。

连续治疗8周后，观察各组动物的一般情况，进行口服糖耐量试验(OGTT)及采用16S rRNA基因扩增子测序比较各组大鼠的肠道菌群。

研究姜黄素对脂多糖(lipopolysaccharide,LPS)诱导糖尿病大鼠肠道菌群的影响。

结果表明：姜黄素可改善LPS所致糖尿病大鼠的多饮、多食等症状并对糖耐量有明显的改善(p<0.05)。

LPS诱导的糖尿病大鼠肠道中提升的Melainabacteria含量可被姜黄素灌服降低(p<0.05)。

本研究揭示姜黄素具有降低LPS诱导的糖尿病大鼠血糖的作用，其机制可能与调节肠道微生态有关。

关键词姜黄素,2型糖尿病,肠道菌群,脂多糖,Wistar大鼠Curcumin May Improve the LPS Induced Type2Diabetes by Adjusting Gut Bacterium Population in Wistar RatLiu Xidi1Xia Ning2*Liang Yuzhen11The First Affiliated Hospital of Guangxi Medical University,Nanning,530021;2Department of Health of Guangxi Zhuang Autonomous Region, Nanning,530021*Corresponding author,xianing12@DOI:10.13417/j.gab.033.000970Abstract Curcumin is kind of Polyphenols or pigments extracted from curcuma zingileraceae plants with great medicinal effects.In present study,thirty healthy male Wistar rats were randomly divided into three groups,a normal control group(n=10),a model group(n=10),and an curcumin treated group(n=10).The rat model of diabetic were made by LPS induced with(300μg·kg·-1day·-1)subcutaneous injection in8weeks.The symptom of diabetes was di-agnosed while the fasting blood glucose(FBG)level reached11.1mmol/L or more,and then the groups were treated with curcumin200mg/(kg·-1day·-1)by gavage.After the treatment in eight weeks later,we observed and recorded the general status and symptom of the Wistar rats,and performed the oral glucose tolerance test(OGTT)as well.Rat gut microbiome profiles by using16S rRNA amplicon sequencing were also carried out in this research in order to assess the effect of curcumin on the gut microbiome of the Wistar rats with LPS induced type2diabets.The results showed that the curcumin could improve the endurance of the blood sugar in the rat models with diabetes caused by lipopolysaccharide(p<0.05).The abundance of Melainabacteria in the gut of insulin resistance rats could be decreased by curcumin oral administration(p<0.05).This study might reveal that the curcumin could improve glucose tolerance in lipopolysaccharide induced diabetic rats,which proposed to modulate intestinal microecologi-cal imbalance.Keywords Curcumin,Type2diabete,Gut microbiome,Lipopolysaccharide,Wistar rat基金项目：本研究由国家自然科学基金(81060070)和广西自然科学基金(2012GXNSFAA053083)共同资助基因组学与应用生物学Genomics and Applied Biology糖尿病是一组以高血糖及胰岛素抵抗为主要特征的慢性代谢性疾病。

罗格列酮对初发2型糖尿病患者糖脂代谢及胰岛素敏感性的影响【摘要】目的：探讨罗格列酮对初发2型糖尿病患者糖脂代谢及胰岛素敏感性的影响。

方法：80例2型糖尿病患者，随机分成试验组（罗格列酮组）和对照组（二甲双胍组），每组40人，研究时间16周。

观察两组患者治疗前后fbg、hba1c、心血管危险因子hs-crp等以及脂肪细胞因子水平和胰岛素抵抗的变化。

结果：治疗后，两组患者fpg、hba1c、fins、homa-ir、抵抗素、瘦素、hsc-rp 值均明显降低（po.05），有可比性（表1）。

1.2 方法采用随机、双盲法。

所有入选对象随机分成试验组和对照组，每组40人。

两组在性别、年龄、体重指数（bmi）、血脂、血压等方面无统计学差异。

用药方法及剂量：试验组，罗格列酮钠（商品名：太罗。

规格：4 mg/片，四川太极集团涪陵制药厂生产）4mg/d，qd；对照组，二甲双胍0.25，tid。

如无特殊情况，不干涉原来的降压、降脂治疗。

共随访16周。

所有患者在治疗前后均详细测量fpg、fins、胰岛素抵抗指数（homa-ir）、hba1c、bmi、腰围、体重、血脂谱、肝功能、血常规等，以及抵抗素、瘦素、高敏c反应蛋白（hscrp）等的水平。

homa-ir=（ fins×fpg）/22.5。

1.3 统计学处理所有数据以表示，采用spss10.0统计软件，对治疗前后采用配对t检验。

p<0.05认为差异有统计学意义。

2 结果2.1 治疗效果两组治疗后fpg、hba1c值均明显降低（p<0.05），试验组tg、tc、ldl-c下降，hdl-c升高，而对照组不明显（表2）；试验组fins、homa-ir、抵抗素、瘦素、hsc-rp值均明显降低（p<0.05），对照组不明显，两组间比较，试验组更优于对照组（p<0.05）（表3）。

2.2 不良事件本研究有2例口服二甲双胍出现轻微胃肠道反应，但未退出试验。

专升本考试：2022英语真题及答案(2)共432道题1、第32题的答案是( )（单选题）A. CrowdedB. extensiveC. largeD. bare试题答案：A2、Which event brought the booming of American candy bar business?（单选题）A. The adding of new materials．B. The demand in the army during WWI．C. The purchase of new machines．D. The appearance of smaller candy bars．试题答案：B3、第33题的答案是( )（单选题）A. reformB. destructionC. SupportD. discovery试题答案：B4、根据下面材料回答{TSE}题： Passage one I talk to strangers for a living and love the challenge of getting their stories published in newspapers．I've been married for years，but until six months ago，I could be a typical absent—minded husband．Often l was just nodding when l was supposed to．When my wife asked，“ Didyou even hear what I just said?”1 would defensively say，“of course I did!” In January，I began to lose my voice．Doctors told me I needed surgery，or my throat would be permanently damaged．Total silence would be required for the first few weeks of my recovery． Two hours after the surgery，my eyes filled with tears as my two-year-old son looked puzzled because l wouldn’t answer his questions．I wanted to talk but couldn’t．Luckily，I'd recorded myself reading some of his favorite books．That would come in handy the next couple of weeks． It had never left．I'd just stopped noticing．I found myself understanding her better on topics I'd previously dismissed as “things I just don’t get as a guy”．I also realized my son wasn’t just talking nonstop but that he often had thoughtful things to say．Even while walking my dog in the woods near our home，I began hearing pleasant patterns in birdsongs．Before my surgery，I'd have spent those walks on my phone． After several weeks，I was fully recovered. Conversation in our house is better now，not because I'm talking more．I’m just listening better and becoming less and less surprised that I like what I hear． {TS}According to the passage，the author is most likely a __________ ．（单选题）A. driverB. teacherC. doctorD. Journalist试题答案：D5、What might be the best title of the passage?（单选题）A. Tea-Drinking TipsB. Lifestyle Secrets of IkariansC. Tea-Drinking Ceremony in OkinawaD. Blue Zones Solutions试题答案：A6、Jonathan and Joe left the house to go for after supper．（单选题）A. walkB. the walkC. walksD. a walk试题答案：D7、第29题的答案是( )（单选题）A. thatB. whoC. whatD. as试题答案：A8、请填写最佳答案（）（单选题）试题答案：F9、第34题的答案是( )（单选题）A. improvementsB. ChangesC. protectionD. development试题答案：B10、第27题答案是（）（单选题）A. causeB. 1eaveC. disturbD. 1ead试题答案：B11、根据以下材料，回答{TSE}题 Insomnia, or "poor sleep",can have bad effects on a person´s health and general well-being. It can 21onboth our physical and mental health and can lead to other health 22 Insomnia can be traced to many differentreasons,but what is 23to ninny sufferers is their inability to relax fully and "switch the mind 24Constantthoughts, 25 around and around in the mind, moving from one 26 tothe next, prevent stillness and peace and 27 a sufferer extremely tired. In order to treat insomnia 28, it isfirst necessary to allow a sufferertore-experience 29 real relaxation feel like.It’s almost as though they´ve forgotten how to relax. Once this hasbeen 30 by the brain, then fast and effective 31 can be made to reeducatethe unconscious towards allowing the person to relax 32 and to allow anatural state of sleep to 33 Hypnotherapy(催眠疗法) is one of the fastest and most effective ways of 34 this goalfor long-lasting results. Sleeping pills, if used at all, shouldonly be a short-term 35as their effect is soon reduced and their side effectscan be deep and far-reaching. {TS}第21题答案是（）（单选题）A. harmB. affectC. changeD. impact试题答案：D12、第31题的答案是( )（单选题）A. tooB. onceC. againD. also试题答案：D13、Only by telling the truth___________win the trust and support of your friends．（单选题）A. you canB. can youC. you didD. did you试题答案：B14、Jason made sharp comments on Mary’s idea，but he didn’t mean __________ her．（单选题）A. HurtB. HurtingC. to hurtD. to be hurt试题答案：C15、第60题应填( )（单选题）试题答案：B16、第25题应选( )（单选题）A. makes upB. focuses onC. refers toD. consists of试题答案：D17、第29题答案是（）（单选题）A. ifB. howC. whereD. what试题答案：D18、根据下面材料回答{TSE}题： Passage one I talk to strangers for a living and love the challenge of getting their stories published in newspapers．I've been married for years，but until six months ago，I could be a typical absent—minded husband．Often l was just nodding when l was supposed to．When my wife asked，“ Did you even hear what I just said?”1 would defensively say，“of course I did!” InJanuary，I began to lose my voice．Doctors told me I needed surgery，or my throat would be permanently damaged．Total silence would be required for the first few weeks of my recovery． Two hours after the surgery，my eyes filled with tears as my two-year-old son looked puzzled because l wouldn’t answer his questions．I wanted to talk but couldn’t．Luckily，I'd recorded myself reading some of his favorite books．That would come in handy the next couple of weeks． It had never left．I'd just stopped noticing．I found myself understanding her better on topics I'd previously dismissed as “things I just don’t get as a guy”．I also realized my son wasn’t just talking nonstop but that he often had thoughtful things to say．Even while walking my dog in the woods near our home，I began hearing pleasant patterns in birdsongs．Before my surgery，I'd have spent those walks on my phone． After several weeks，I was fully recovered. Conversation in our house is better now，not because I'm talking more．I’m just listening better and becoming less and less surprised that I like what I hear． {TS}According to the passage，the author is most likely a __________ ．（单选题）A. driverB. teacherC. doctorD. Journalist试题答案：D19、What is the passage mainly about?（单选题）A. Selection process of rescue dogs．B. Qualities and training of rescue dogs．C. Risks rescue dogs are faced with．D. Types of tasks rescue dogs can perform．试题答案：B20、The old man，together with his neighbors，__________ the performance when it began to rain．（单选题）A. was enjoyingB. were enjoyingC. has enjoyedD. have enjoyed试题答案：A21、第23题的答案是( )（单选题）A. for exampleB. in additionC. at lastD. after all试题答案：A22、第33题应选( )（单选题）A. racesB. regionsC. mindD. mankind试题答案：D23、The carpet has so many stains on it thatit needs__________（单选题）A. replaceB. to replaceC. being replacedD. to be replaced试题答案：D24、选出下列选项中划线部分读音不同的选项（）。

糖尿病合并冠心病男性患者血清睾酮水平变化（作者:___________单位: ___________邮编: ___________）作者：金玎陈丽珠郑玉云郭晓华【摘要】目的：观察糖尿病合并冠心病的男性患者血清睾酮水平。

方法：入选经冠状动脉造影证实的冠心病患者83例,包括糖尿病19例和非糖尿病64例。

以化学发光免疫法（CLIA）检测血清睾酮(testosterone，T)、雌二醇(estradiol，E2)水平。

结果：糖尿病合并冠心病者T水平低于无糖尿病的冠心病者（3.12±1.42μg/L与3.93±1.13μg/L,t=2.304，P0.05）。

结论：合并糖尿病的男性冠心病者T水平低于非糖尿病冠心病者。

【关键词】糖尿病；睾酮；冠状动脉造影；男性；冠心病Abstract Objective:To observe the changes in serum testosterone of the male CHD patients with Diabetes Mellitus.Methods: 83 male CHD patients confirmed by coronary angiography were distributed into diabetes mellitus group (n=19) and non-diabetes mellitus group (n=64).The serum concentrationof testosterone(T) and estradiol(E2) was determined by the Chemiluminesent Immunoassay Assay（CLIA）.Results: The CHD patients with diabetes mellitus had significantly lower levels of serum T than those without diabetes mellitus(3.12±1.42 vs 3.93±1.13μg/L, t=2.304 P0.05).Conclusion: Serum T level in the male CHD patients with diabetes mellitus is remarkably lower than that in those without suffering from diabetes mellitus.Key words Diabetes mellitus; Testosterone; Coronary angiography; Male coronary heart disease糖尿病是一种常见的全身性代谢紊乱性疾病，大量的流行病学资料证明糖尿病是冠心病的独立危险因素，并已被国内外学者所公认。

男性2型糖尿病病人血糖与雄激素水平的相关性作者：沈洁廖小平来源：《中外医疗》2019年第04期[摘要] 目的通过研究男性2型糖尿病病人的糖化血红蛋白与血清雄激素水平的相关性，探讨男性2型糖尿病患者血糖与其血清雄激素水平的相关性。

方法方便收集2015年1月—2017年12月上海市嘉定区安亭医院收治的110例男性2型糖尿病患者，测定其糖化血红蛋白（HbA1c）水平，血清睾酮（T）水平、甘油三酯（TG）、总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）。

以T<14 nmol/L以及T≥14 nmmol/L分为两组，比较两组间的TC、TG、HbA1c、LDL-C、年龄的差异是否有统计学意义，并以T为因变量，TC、TG、HbA1c、LDL-C、年龄为自变量进行Logistic回归分析。

结果 T<14 nmol/L组的HbA1c平均值为9.76%，平均年龄为57.64岁，T≥14 nmmol/L组的HbA1c平均值为8.28%，平均年龄为52.51岁，两组间的血清HbA1c以及年龄的差异有统计学意义（P[关键词] 睾酮;糖化血红蛋白;2型糖尿病;男性;血糖[中图分类号] R587 ; ; ; ; ;[文献标识码] A ; ; ; ; ;[文章编号] 1674-0742（2019）02（a）-0032-03[Abstract] Objective To investigate the correlation between glycosylated hemoglobin and serum androgen levels in male patients with type 2 diabetes， and to explore the correlation between blood glucose and serum androgen levels in male patients with type 2 diabetes. Methods From January 2015 to December 2017， 110 male patients with type 2 diabetes were convenient collected from Anting Hospital of Jiading District， Shanghai， and their glycated hemoglobin （HbA1c） levels， serum testosterone （T） levels， triglycerides （TG）， total cholesterol （TC）， low density lipoprotein cholesterol （LDL-C）. Based on the T<14 nmol/L and T≥14nmmol/L， these patients were divided into two groups. The difference of TC， TG， HbA1c， LDL-C and age between the two groups was statistically significant， and T was the dependent variable. Logistic regression analysis was performed for TG， HbA1c， LDL-C， and age. Results The mean value of HbA1c in the T<14 nmol/L group was 9.76%， the mean age was 57.64 years old， the mean value of HbA1c in the T≥14 nmmol/L group was 8.28%， and the mean age was 52.51 years old. Serum HbA1c and age between the two groups. The difference was statistically significant （P[Key words] Testosterone; Glycated hemoglobin; Type 2 diabetes; Male; Blood glucoseHbA1c可以反映糖尿病患者的近2～3个月的平均血糖水平，其浓度不会受即时血糖的干扰，目前已被世界卫生组织（WHO）列为糖尿病的一个诊断标准[1]，是评价治疗方案的有效性和血糖管理的合理性的金标准。

药学通识英语试题及答案一、选择题（每题2分，共20分）1. Which of the following is not a primary function of drugs?A. DiagnosisB. TreatmentC. PreventionD. Rehabilitation答案：A2. The term "pharmacology" refers to the study of:A. The effects of drugs on living organismsB. The synthesis of new drugsC. The distribution of drugs in the marketD. The legal regulations of drugs答案：A3. The most common route of drug administration is:A. OralB. IntravenousC. IntramuscularD. Topical答案：A4. Which of the following is a side effect of a drug?A. The intended therapeutic effectB. An effect that is harmful and unintendedC. The effect of the drug on a different organD. The effect of the drug on a different disease答案：B5. The half-life of a drug refers to:A. The time it takes for the drug to be completely eliminated from the bodyB. The time it takes for the drug's concentration to decrease by halfC. The time it takes for the drug to reach its maximum concentrationD. The time it takes for the drug to be absorbed into the bloodstream答案：B6. The bioavailability of a drug is:A. The percentage of the drug that is absorbed into the bloodstreamB. The percentage of the drug that is excreted unchangedC. The percentage of the drug that is metabolized by theliverD. The percentage of the drug that is stored in the fat tissues答案：A7. The therapeutic index of a drug is an indicator of:A. The drug's effectivenessB. The drug's safetyC. The drug's cost-effectivenessD. The drug's duration of action答案：B8. A drug's pharmacokinetics involves the study of:A. How the body affects the drugB. How the drug affects the bodyC. How the drug is synthesizedD. How the drug is regulated by the government答案：A9. The first-pass metabolism refers to:A. The metabolism of a drug after it is absorbed into the bloodstreamB. The metabolism of a drug before it enters the bloodstreamC. The metabolism of a drug after it is excreted from the bodyD. The metabolism of a drug after it is distributed to the tissues答案：B10. The term "drug interaction" refers to:A. The combined effect of two or more drugsB. The effect of one drug on the action of another drugC. The effect of a drug on the patient's behaviorD. The effect of a drug on the patient's diet答案：B二、填空题（每题2分，共20分）1. The study of the effects of drugs on living organisms is called __________.答案：pharmacology2. The intended therapeutic effect of a drug is known as its __________.答案：pharmacological effect3. The process by which a drug is absorbed into the bloodstream is called __________.答案：absorption4. A drug that is taken orally and then undergoes metabolism in the liver before entering the bloodstream is subject to__________.答案：first-pass metabolism5. The time it takes for the drug's concentration to decrease by half is known as the drug's __________.答案：half-life6. The percentage of the drug that is absorbed into the bloodstream is referred to as the drug's __________.答案：bioavailability7. The study of how the body affects the drug is known as the pharmacokinetics of the drug, while the study of how the drug affects the body is known as the __________.答案：pharmacodynamics8. A drug's safety is indicated by its __________.答案：therapeutic index9. The combined effect of two or more drugs is known as a__________.答案：drug interaction10. The unintended harmful effect of a drug is called a(n)__________.答案：side effect三、简答题（每题10分，共40分）1. Explain the difference between the pharmacokinetics and pharmacodynamics of a drug.答案：Pharmacokinetics is the study of how the body affects the drug, including absorption, distribution, metabolism, and excretion. Pharmacodynamics, on the other hand, is the study of how the drug affects the body, including the drug's mechanism of action and its effects on physiological functions.2. Describe the significance of a drug's half-life inclinical practice.答案：The half-life of a drug is significant in clinical practice as it determines the frequency of drug administration. A shorter half-life may require more frequent dosing, while a longer half-life allows for less frequent dosing. It also。

内分泌治疗英语Endocrine TherapyThe endocrine system is a complex network of glands and hormones that play a crucial role in regulating various physiological processes within the human body. Hormones, the chemical messengers produced by these glands, are responsible for maintaining homeostasis, controlling growth and development, and influencing a wide range of bodily functions, from metabolism to mood. When the endocrine system malfunctions, it can lead to a variety of health issues, ranging from hormonal imbalances to chronic diseases. In such cases, endocrine therapy emerges as a vital treatment approach, aiming to restore the delicate balance of hormones and alleviate the associated symptoms.Endocrine therapy, also known as hormone therapy, is a medical treatment that involves the administration of synthetic or natural hormones to address hormonal imbalances or to target specific hormone-dependent conditions. This therapeutic approach is particularly effective in the management of various endocrine-related disorders, including thyroid dysfunction, diabetes, and certain types of cancer.One of the primary applications of endocrine therapy is in the treatment of thyroid disorders. The thyroid gland, located in the neck, is responsible for producing hormones that regulate metabolism, body temperature, and other vital functions. When the thyroid gland becomes underactive (hypothyroidism) or overactive (hyperthyroidism), it can lead to a range of symptoms, such as fatigue, weight changes, and mood disturbances. Endocrine therapy, typically in the form of synthetic thyroid hormones (e.g., levothyroxine), can be used to restore the proper balance of thyroid hormones, effectively managing the symptoms and preventing long-term complications.Another significant application of endocrine therapy is in the treatment of diabetes, a chronic condition characterized by thebody's inability to regulate blood sugar levels effectively. In type 1 diabetes, the pancreas fails to produce insulin, a hormone essential for glucose metabolism. In type 2 diabetes, the body becomes resistant to the effects of insulin. Endocrine therapy in the form of insulin administration or the use of other antidiabetic medications (e.g., metformin, GLP-1 agonists) can help maintain healthy blood sugar levels, reducing the risk of diabetic complications such as nerve damage, kidney disease, and cardiovascular problems.Endocrine therapy also plays a crucial role in the management ofhormone-dependent cancers, such as breast cancer and prostate cancer. These types of cancers are fueled by the presence of specific hormones, which promote the growth and proliferation of cancer cells. Endocrine therapy in this context aims to disrupt the cancer's dependence on hormones, either by blocking the production or action of these hormones or by directly targeting the cancer cells. This approach can be highly effective in slowing the progression of the disease, improving patient outcomes, and enhancing quality of life.In the case of breast cancer, endocrine therapy may involve the use of selective estrogen receptor modulators (SERMs), such as tamoxifen, or aromatase inhibitors, which block the conversion of androgens to estrogens. These medications can significantly reduce the risk of recurrence and improve survival rates in patients with hormone-receptor-positive breast cancer. Similarly, in prostate cancer, endocrine therapy may involve the use of androgen-deprivation therapies, which suppress the production or action of testosterone, the primary male sex hormone.Beyond these well-established applications, endocrine therapy is also being explored for the management of other health conditions, such as polycystic ovary syndrome (PCOS), infertility, and certain endocrine-related mental health disorders. In PCOS, for instance, endocrine therapy may involve the use of oral contraceptives orinsulin-sensitizing medications to regulate hormonal imbalances and alleviate symptoms like irregular menstrual cycles, acne, and excessive hair growth.The success of endocrine therapy is largely dependent on the accurate diagnosis and monitoring of the underlying endocrine condition. Healthcare providers, such as endocrinologists, work closely with patients to develop personalized treatment plans, which may involve a combination of medication, lifestyle modifications, and regular monitoring of hormone levels. Patients play a crucial role in the treatment process, as adherence to the prescribed regimen and regular follow-up appointments are essential for achieving optimal outcomes.In conclusion, endocrine therapy is a vital component of modern healthcare, providing effective solutions for a wide range of endocrine-related disorders. By leveraging the power of hormones and targeted interventions, healthcare professionals can help patients restore hormonal balance, manage chronic conditions, and improve their overall health and well-being. As research in this field continues to evolve, the potential of endocrine therapy to address complex health challenges is expected to grow, offering new hope and improved quality of life for those affected by endocrine system disorders.。

睾酮对血管功能及动脉粥样硬化的影响及其机制的研究进展赵锐;李艳【摘要】血清睾酮浓度降低与男性心血管疾病的发生密切相关.睾酮能够调节血清胆固醇代谢、内皮细胞功能紊乱、炎症等与动脉粥样硬化相关的危险因素,降低心血管事件的发生,并具有舒张血管的作用,还可增加心肌供血,改善心绞痛患者的临床症状.此外,睾酮还具有抗动脉粥样硬化的作用,睾酮的缺乏会促使早期动脉粥样硬化形成.本文就睾酮对血管功能及动脉粥样硬化的影响及其机制的研究进展进行综述,为睾酮作为激素替代疗法治疗或预防心血管疾病提供理论参考.【期刊名称】《广西医学》【年(卷),期】2016(038)004【总页数】4页(P533-536)【关键词】睾酮;血管;功能;动脉粥样硬化;综述【作者】赵锐;李艳【作者单位】武汉大学人民医院检验科,武汉市430060;武汉大学人民医院检验科,武汉市430060【正文语种】中文【中图分类】R543.5睾酮由睾丸间质细胞及肾上腺皮质合成并分泌，受下丘脑-垂体的负反馈系统所调控，血液循环中高达80%的睾酮与性激素结合球蛋白结合，游离睾酮占2%。

睾酮主要在肝脏代谢生成17-酮类衍生物，从尿液中排出。

睾酮在人体内发挥重要的生理功能：(1)促进精子的生成和成熟；(2)刺激男性生殖系统的发育；(3)维持男性正常性功能；(4)促进骨骼、肌肉生长和蛋白质合成。

研究发现，睾酮可调节血脂、血糖、同型半胱氨酸等物质代谢，并具有抗动脉粥样硬化(atherosclerosis，AS)的作用，其水平与男性冠心病(coronary heart disease，CHD)的发病率密切相关[1]。

有研究认为，血清睾酮水平降低可作为心血管疾病的独立危险因素[2]。

雄激素阻断疗法治疗前列腺癌，增加了患者因糖尿病以及CHD等心血管相关疾病死亡的风险[3]。

动物实验发现，切除睾丸的雄性动物模型AS斑块形成增多，及时补充睾酮后AS斑块明显减少，表明睾酮具有抗AS功能[4]。

Administration of L-carnitine and mildronateimproves endothelial function and decreasesmortality in hypertensive Dahl ratsReinis Vilskersts1,Janis Kuka1,2,Baiba Svalbe1,3,Helena Cirule1,Edgars Liepinsh1,Solveiga Grinberga1,Ivars Kalvinsh1,Maija Dambrova1Latvian Institute of Organic Synthesis,Aizkraukles21,Riga,LV-1006,LatviaRigas Stradins University,Dzirciema16,Riga,LV-1007,Latvia!University of Latvia,Raina Blvd.19,Riga,LV-1050,LatviaCorrespondence:Reinis Vilskersts,e-mail:Reinis.Vilskersts@biomed.lu.lvAbstract:Hypertension is a well established risk factor for the development of cardiovascular diseases and increased mortality.This study was performed to investigate the effects of the administration of L-carnitine or mildronate,an inhibitor of L-carnitine biosynthesis,or their combination on the development of hypertension-related complications in Dahl salt-sensitive(DS)rats fed with a high salt diet.Male DS rats were fed laboratory chow containing8%NaCl from7weeks of age.Experimental animals were divided into five groups and treated for8weeks with vehicle(water;n=10),L-carnitine(100mg/kg,n=10),mildronate(100mg/kg,n=10)or a combination of L-carnitine and mildronate at the doses above(n=10).During the experiment,control group animals continued to consume a diet with normal salt content.Administration of the combination significantly improved the survival rate for50%of the population.None of the tested compounds or their combination influenced high salt intake-induced hypertension,while treatment with mildronate and the com-bination for8weeks significantly decreased resting heart rate by12%and10%,respectively.Feeding with high salt diet had no influ-ence on systolic function of the heart,but it induced thickening of the ventricular walls and development of heart hypertrophy that was not improved by the administration of tested compounds.In addition,administration of the combination attenuated the development of endothelial dysfunction in isolated aortic rings.In conclusion,our results demonstrate that treatment with a combination of L-carnitine and mildronate is protective against hypertension-induced complications in an experimental model of salt-induced hypertension. Key words:mildronate;L-carnitine;Dahl salt-sensitive rats;hypertension;endothelial dysfunctionAbbreviations:BW–body weight,DS rats–Dahl salt-sensitive rats,GBB–g-butyrobetaine,IVSd–interventricular septal thickness at end-diastole,IVSs–interventricular septal thickness at end-systole,LVEF–left ventricular ejection frac-tion;LVFS–left ventricular fractional shortening,LVIDd–left ventricular internal dimension at end-diastole,LVIDs–left ventricular internal dimension at end-systole,LVPWd–left ventricular posterior wall thickness at end-diastole,LVPWs–left ventricular posterior wall thickness at end-systole,LVW–left ventricular weight,UPLC/MS/MS–ultra performance liq-uid chromatography-tandem mass spectrometry IntroductionHypertension is a well established risk factor for myo-cardial infarction and stroke[13],the development of endothelial dysfunction[17]and atherosclerosis[4]. Large clinical trials have shown that a reduction of ar-terial blood pressure attenuates the development of complications of hypertension[23,45].Nevertheless, it has recently been demonstrated that drugs that doPharmacological Reports 2011,63,752 762ISSN1734-1140Copyright©2011by Institute of Pharmacology Polish Academy of Sciencesnot have a direct impact on blood pressure were also able to attenuate the development of complications of hypertension[21,44];thus,a reduction of blood pres-sure is not the only therapeutic strategy to prevent complications of hypertension.Some experimental data have indicated that treatment with L-carnitine has a certain impact on the development of hyperten-sion-related complications[5,9].L-Carnitine is an amino acid derivative that is essen-tial for the transport of long-chain fatty acids from the cytoplasm into sites of b-oxidation in the mitochondria [12].Additional supplementation of L-carnitine has been shown to be beneficial in different pathologies that could be induced by hypertension.Results from several studies have proven the beneficial effects of the administration of L-carnitine in the case of cardio-vascular pathologies[10],and treatment with L-carnitine has attenuated the development of athero-sclerosis[39]and reduced endothelial dysfunction [42].Recent experimental data have also shown that in some experimental models,the administration of L-carnitine was able to normalize elevated systolic blood pressure[29,36].Mildronate[3-(2,2,2-trimethylhydrazinium)propi-onate]is an inhibitor of biosynthesis of L-carnitine [43]and its reabsorption in kidneys[22];thus,the ad-ministration of mildronate decreases the L-carnitine content in plasma and different tissues[16,24].The pharmacological effects of mildronate are based on its regulatory effect on L-carnitine concentration,with subsequent changes to downstream pathways of en-ergy metabolism[8,25].Similar to L-carnitine,mil-dronate treatment has shown an impact on complica-tions and diseases that could be induced by hyperten-sion.The anti-atherosclerotic and angioprotective effects of mildronate have been proven in experimen-tal models of atherosclerosis and diabetes[26,48]. Moreover,mildronate treatment protected myocar-dium against ischemia-reperfusion-induced damage [24,41]and improved cardiac function in experimen-tal models of heart failure[2,15].The aim of the present study was to investigate whether the administration of L-carnitine,mildronate or their combination is protective against hyperten-sion-induced complications.Effects on the develop-ment of hypertension-induced complications were tested in Dahl salt-sensitive(DS)rats fed with a high salt diet,because during a high salt load,DS rats de-velop marked hypertension with cardiovascular com-plications[27,51].Our results provide the first experimental evidence that treatment with a combination of mildronate and L-carnitine is protective against hypertension-induced endothelial dysfunction,and at the same time de-creases overall morality.Materials and MethodsChemicalsMildronate dihydrate was obtained from JSC Grindeks (Latvia).Ketamine hydrochloride solution(Bioketan) was from Vetoquinol Biowet(Poland)and xylazine hydrochloride solution(Seton2%)was from Labora-torios Calier(Spain).Acetonitrile and methanol were obtained from Merck(Germany).(2,2-Dimethyl-2-prop-1-yl-hydrazinium)propionate was prepared in-house.L-Carnitine,sodium chloride,potassium chlo-ride,calcium chloride dihydrate,magnesium chloride hexahydrate,sodium hydrogencarbonate,potassium dihydrogenphosphate,glucose,ethylenediaminetetra-acetic acid(EDTA),L-phenylephrine hydrochloride, acetylcholine chloride,ammonium acetate and so-dium pyruvate were purchased from Sigma(USA).Experimental animals and protocolThe experimental procedures were carried out in ac-cordance with the guidelines of the European Com-munity and local laws and policies and were approved by the Latvian Animal Protection Ethical Committee of the Food and Veterinary Service,Riga,Latvia.Six week old male DS rats were purchased from Charles River(Germany)and maintained under con-trolled conditions of light,temperature and humidity. After a week of adaptation to a new environment,the rats were divided into5groups according to treatment and diet:rats of the first group received a normal salt (0.3%NaCl)diet and drinking water(NS group);rats of the second group received a high salt(8%NaCl) diet(Special Diets Services,Great Britain)and drink-ing water(HS group);rats of the third group received a high salt(8%NaCl)diet and L-carnitine at a dose of 100mg/kg together with drinking water(HS+C100 group);rats of the fourth group received a high salt diet and mildronate at a dose of100mg/kg together with drinking water(HS+M100group);rats of the L-carnitine and mildronate is protective in hypertensionReinis Vilskersts et al.fifth group received a high salt diet and mildronate and L-carnitine,both at doses of100mg/kg together with drinking water(HS+CM group).Experimental animals received the experimental diets and test com-pounds for8weeks.The dosing of mildronate,L-car-nitine or their combination was confirmed by measur-ing the consumption of drinking water every2days and adjusting the concentration of supplemented sub-stances.At the beginning of the experiment,when DS rats started to receive diet with high salt load,the concen-trations of tested compounds in the drinking water were as follows:L-carnitine1.004g/l;mildronate di-hydrate1.406g/l and in the drinking water of the combination group:L-carnitine0.876g/l and mil-dronate dihydrate1.086g/l.At the end of the study when experimental animals consumed larger amounts of drinking water because of high salt diet,solutions of the tested compound had to be more diluted.Thus, at the end of the experiment concentrations of tested drugs in the drinking water were as follows:L-carnitine 0.202g/l;mildronate dihydrate0.218g/l and in the drinking water of the combination group:L-carnitine 0.199g/l and mildronate dihydrate0.246g/l.Physiological and echocardiographic measure-ments in DS rats were performed at the beginning of the experiment and4and8weeks after the treatment. At15weeks of age,DS rats were sacrificed under an-esthesia.Plasma was taken for the quantitative deter-mination of L-carnitine,g-butyrobetaine(GBB)and mildronate.The thoracic aorta was dissected for organ chamber experiments as described below.All analy-ses were performed by an observer blinded to the treatment group.Physiological and echocardiographicmeasurementsSystolic blood pressure and heart rate were measured in conscious DS rats at the beginning of the experi-ment and after4and8weeks of administration of the test compounds using a Non-Invasive Blood Pressure Controller ML125connected to PowerLab8/30sys-tem(ADInstruments)and a PC.Transthoracic echocardiographic studies were per-formed on DS rats with iE33ultrasonograph using linear L15-7io transducer(Philips Ultrasound Inc., USA).Rats were lightly anesthetized with an intrape-ritoneal injection of ketamine and xylazine(50and 10mg/kg)and were held in the decubitus position.The rats were allowed to breathe spontaneously dur-ing the procedure.M-mode tracings of the left ventri-cle were recorded at the papillary muscle level to measure left ventricular weight(LVW),interventricu-lar septal thickness at end-diastole(IVSd),interven-tricular septal thickness at end-systole(IVSs),left ventricular posterior wall thickness at end-diastole (LVPWd),left ventricular posterior wall thickness at end-systole(LVPWs),left ventricular internal dimen-sion at end-diastole(LVIDd),left ventricular internal dimension at end-systole(LVIDs),left ventricular ejection fraction(LVEF)and left ventricular frac-tional shortening(LVFS).The average of three values for each rat was calculated.Organ chamber experimentEndothelial function was examined in aortic rings us-ing an organ chamber bath as described previously [3].In brief,the thoracic aorta was excised,immersed in ice-cold Krebs-Henseleit(K-H)buffer(content in mmol/l:NaCl118,CaCl22.5,MgCl 1.64,NaHCO!24.88,KH PO"1.18,glucose10.0,EDTA0.05)pH 7.4and cleaned of fatty and connective tissue.The aorta was cut into3mm long aortic rings that were mounted between two stainless steel hooks in oxygen-ated K-H buffer(pH7.4,37°C).The passive tension was fixed at40mN.After a period of equilibration (1h),the maximal contractile function was assessed by application of60mM KCl.The aortic rings were then precontracted to70%of maximal contraction with phenylephrine and the cumulative response to acetylcholine(10` –10`#M)was assessed.Relaxa-tion of aortic rings to acetylcholine was expressed as a percent of phenylephrine-induced constriction.Determination of L-carnitine,GBBand mildronateThe determination of L-carnitine,GBB and mildronate concentrations in the blood plasma samples was per-formed by ultra performance liquid chromatography-tandem mass spectrometry(UPLC/MS/MS)in positive ion electrospray mode,as described previously[7].Data analysisData were expressed as the mean±SEM.Statistical calculations were performed using the GraphPad Prism3.0software package(USA).Survival curveswere constructed for each group using the Kaplan-Meier method,and comparison of survival distribu-tions among the groups was performed using the log-rank test.For the comparison of the response of aortic rings to acetylcholine,two-way ANOV A with the Bonferroni post-hoc test was performed.In case of other parameters differences between means were evaluated by one way ANOV A followed by a Mann-Whitney U test.The differences were considered sig-nificant when p<0.05.ResultsSurvival rateKaplan-Meier analysis revealed that the survival rate of rats receiving a diet with high salt content(HS group)was markedly reduced compared with that of animals from the NS group(Fig.1).In the HS group after8weeks of treatment,the survival rate was30% (3of10animals),while there were no lethal cases in the NS group.Administration of a combination of L-carnitine and mildronate significantly increased the survival rate compared with the HS group,and after8 weeks of treatment,the HS+CM group survival rate was80%(8of10animals).The obtained data showed that there was no statistically significant increase in survival rate in the HS+C100(survived8of10ani-mals)and HS+M100(survived8of10animals)groups compared with the HS group although the number of survived animals were similar as in HS+CM group.Systolic blood pressure and heart rateAt the beginning of the experiment,there were no sig-nificant differences among the mean systolic blood pressure of animals in all of the experimental groups (Fig.2A),and the average systolic blood pressure was about125mmHg.After4and8weeks of treatment, the systolic blood pressure of DS rats receiving a high salt diet had increased to above170mmHg.As can be seen in Figure2A,none of the test compounds or their combination influenced systolic blood pressure dur-ing the experiment.The analysis of heart rate showed that there were no differences among the heart rate of animals of all of the experimental groups before switching to the high salt diet or on the fourth week of the treatment.However,on the eighth week of the ex-periment,the mean heart rate in the HS group was 510±10bpm,but administration of mildronate and a combination of mildronate and L-carnitine de-creased it by12and10%,respectively(Fig.2B).Organ weight and echocardiographic analysis High salt loading in DS rats induced hypertension with the subsequent development of left ventricular hyper-trophy.As can be seen in Table1,significant hypertro-phy of the left ventricle of the heart was found after 4weeks of feeding with the high salt diet.In NS group animals,the left ventricle/body weight index(LVW/ BW)on the fourth and eighth week of the experiment was 2.8±0.1g/kg,while in HS group animals, LVW/BW on the fourth and eighth week of the experi-ment were3.7±0.1g/kg and4.4±0.3g/kg,respec-L-carnitine and mildronate is protective in hypertensionReinis Vilskersts etal. Fig.1.Kaplan-Meier plots of the sur-vival rate of DS rats fed a normal-saltdiet or a high salt diet and treatedeither with vehicle,L-carnitine,mil-dronate or their combination.*p<0.05vs.HS group;**p<0.01vs.HS groupFig.2.Time course of systolic bloodpressure(A)and heart rate(B)of DSrats fed a normal-salt diet(NS)ora high salt diet(HS)and treated eitherwith vehicle,L-carnitine(HS+C100),mildronate(HS+M100)or their combi-nation(HS+CM).Results are pre-sented as the mean±SEM of at least three animals.**p<0.01vs.HS group;***p<0.001vs.HS groupFig. 3.Endothelium-dependent re-laxation to acetylcholine in aortic ringsof DS rats fed a normal-salt diet(NS)ora high salt diet(HS)and treated eitherwith vehicle,L-carnitine(HS+C100),mildronate(HS+M100)or their combi-nation(HS+CM).Results are pre-sented as the mean±SEM of at leastthree animals.*p<0.05vs.HS group;***p<0.001vs.HS grouptively.In addition,renal hypertrophy was observed in experimental animals receiving diet with high salt load. None of the tested compounds or their combination in-fluenced the development of heart or renal hypertro-phy.Moreover,data from our study showed that in DS rats receiving a high salt diet for8weeks,significant pulmonary congestion was still absent(Tab.1).Results from the echocardiographic analysis are summarized in Table2.Feeding experimental animals with diet with high salt load induced thickening of the walls of the left ventricle during diastole as well as systole.None of the tested compounds or their combi-nation influenced thickening of the walls of the left ventricle.Analysis of the systolic parameters of the left ventricle(LVEF and LVFS)revealed that there were no significant differences between mean values of the experimental groups(Tab.2).Endothelium-dependent relaxationto acetylcholine in aortic ringsEndothelium-dependent relaxation to acetylcholine in the aortic rings of HS group animals was significantly impaired compared with the aortic rings of NS group animals(Fig.3).Treatment with a combination of mildronate and L-carnitine improved endothelium-dependent relaxation,but the administration of mil-dronate or L-carnitine alone for8weeks had no im-pact on acetylcholine-induced relaxation(Fig.3).L-carnitine,GBB and mildronate content in rat plasmaThe UPLC/MS/MS analysis of plasma samples revealed that the consumption of diet with high salt load de-creased concentration of L-carnitine nearly2-fold com-pared with that of rats consuming diet with normal salt load(Tab.3).Administration of L-carnitine or combina-tion significantly increased the concentration of L-car-nitine in the plasma,but the treatment with mildronate at the dose of100mg/kg decreased it.The obtained data showed that the concentration of mildronate in plasma samples of HS+CM group animals was three times lower than in HS+M100group animal plasma samples (Tab.3).Moreover,the administration of mildronate, carnitine or their combination significantly increased the concentration of GBB in plasma samples. DiscussionIn the present experimental study,we investigated whether the administration of L-carnitine,mildronate or their combination could attenuate the progression of hypertension-related complications in DS hyper-tensive rats.The obtained results showed that only ad-ministration of a combination consisting of mil-dronate and L-carnitine had a significant effect on mortality and the development of endothelial dys-function compared to a vehicle group with no phar-macotherapy that was consuming the same salt diet.The UPLC/MS/MS analysis of plasma samples re-vealed that experimental animals from HS group had decreased plasma concentration of carnitine com-pared with NS group animals(Tab.3).Differences between both groups could be due to increased excre-tion of urine in HS group animals(data not shown)as L-carnitine and mildronate is protective in hypertensionReinis Vilskersts et al.Tab.1.Body weights and organ to body weight indexesBW (g)LVW/BW(g/kg)Lung/BW(g/kg)Kidney/BW(g/kg)NS group0.week229±6 3.1±0.2––4.week362±7** 2.8±0.1**––8.week408±9** 2.8±0.1** 4.7±0.78.9±0.2** HS group0.week223±5 3.0±0.1––4.week329±7 3.7±0.1––8.week334±19 4.4±0.3 5.5±0.912.9±1.0 HS+M100group0.week224±4 2.9±0.1––4.week340±6 3.7±0.1––8.week342±14 4.6±0.6 5.9±1.012.7±0.7 HS+C100group0.week224±5 3.2±0.1––4.week333±4 3.6±0.1––8.week300±14 5.3±0.3 5.0±0.512.2±0.8 HS+CM group0.week224±5 3.3±0.2––4.week337±9 3.7±0.1––8.week324±15 4.1±0.3 5.9±1.213.4±0.7 The body weight(BW),left ventricle to body weight index(LVB/BW), lung to body weight index(lung/BW)and kidney to body weight index (kidney/BW)of the DS rats before the study and on the fourth and eighth weeks of the experiment.Results are presented as the mean±SEM of at least three animals.**p<0.01vs.HS groupTab.2.Changes in echocardiographic parametersNS group HS group HS+M100group HS+C100group HS+CM group IVSd(mm)0.week 1.39±0.01 1.43±0.04 1.40±0.08 1.39±0.03 1.41±0.034.week 1.69±0.03* 1.90±0.04 1.88±0.05 1.90±0.04 1.92±0.098.week 1.76±0.04* 2.20±0.04 2.32±0.05 2.26±0.06 2.23±0.09IVSs(mm)0.week 2.62±0.02 2.66±0.13 2.42±0.17 2.71±0.16 2.72±0.114.week 2.76±0.05* 3.20±0.08 3.26±0.11 3.27±0.11 3.11±0.068.week 2.92±0.05* 3.68±0.16 3.76±0.16 3.73±0.12 3.45±0.10LVPWd(mm)0.week 1.48±0.04 1.67±0.11 1.54±0.12 1.71±0.16 1.73±0.124.week 1.79±0.06* 2.07±0.06 2.11±0.04 2.05±0.05 2.08±0.048.week 1.87±0.05* 2.29±0.04 2.56±0.13 2.73±0.21 2.39±0.17LVPWs(mm)0.week 2.71±0.27 2.86±0.05 2.74±0.18 3.18±0.20 3.10±0.054.week 2.81±0.11* 3.22±0.11 3.35±0.05 3.29±0.10 3.36±0.128.week 2.94±0.08* 3.73±0.16 3.87±0.16 3.96±0.12 3.72±0.21LVIDd(mm)0.week7.21±0.12 6.84±0.18 6.59±0.14 6.81±0.45 6.77±0.204.week7.85±0.087.86±0.158.02±0.117.85±0.167.90±0.198.week8.05±0.127.88±0.197.42±0.267.01±0.24*7.73±0.17LVIDs(mm)0.week 3.67±0.01 3.61±0.27 3.44±0.01 3.12±0.33 3.37±0.224.week 4.85±0.15 4.72±0.16 4.69±0.15 4.57±0.25 4.68±0.258.week 5.01±0.15 4.30±0.33 4.05±0.31 3.66±0.11 4.39±0.25LVFS(%)0.week50±347±348±154±248±14.week41±240±142±142±242±18.week41±245±441±247±341±2LVEF(%)0.week84±183±383±186±285±34.week75±276±277±177±276±28.week77±280±477±282±277±3 Interventricular septal thickness at end-diastole(IVSd),interventricular septal thickness at end-systole(IVSs),left ventricular posterior wall thickness at end-diastole(LVPWd),left ventricular posterior wall thickness at end-systole(LVPWS),left ventricular internal dimension at end-diastole(LVIDd),left ventricular internal dimension at end-systole(LVIDs),left ventricular fractional shortening(LVFS)and left ventricular ejec-tion fraction(LVEF)of the DS rats before the study and on the fourth and eighth weeks of the experiment.Results are presented as the mean ±SEM of at least three animals.*p<0.05vs.HS groupwell as developing renal injuries evidenced in other studies[11,52]that could result in an increased renal loss of L-carnitine.Previous studies that have investi-gated effects of L-carnitine administration on cardio-vascular diseases,including hypertension-related complications,used at least two times higher dose of L-carnitine[5,9,32].In our study we chose the dose of administered L-carnitine that restored plasma con-centration of carnitine of HS group animals to the level of NS group animals to study the effects of nor-mal carnitine concentration on the development of hypertension-related complications.Administration of mildronate at the dose of100mg/kg induced sig-nificant decrease in plasma L-carnitine concentration that was in accordance with previous studies[24,26]. In addition,simultaneous administration of L-carnitine and mildronate did not restore the plasma concentra-tion of L-carnitine to the level of NS group and plasma concentration of mildronate was lower than found in HS+M100group animals(Tab.3).Lower plasma concentrations of both compounds could be due to similar reabsorbtion pathways[14]and influ-ence of mildronate on the synthesis and reabsorbtion of L-carnitine[22,43].Despite to lower plasma con-centrations of mildronate and L-carnitine in plasma of HS+CM group animals,we found pronounced in-crease in plasma GBB concentration(Tab.3).We sup-pose that the increase in GBB concentration is firstly due to action of mildronate on the concentration of GBB[24]and secondly due to reabsorbed GBB,that could occur as a result of metabolism of L-carnitine [37].Moreover,we observed elevated GBB concen-tration in the plasma of HS+C100group animals,but the increase was not so pronounced probably because of conversion of GBB to L-carnitine[38].Significant differences in the rate of mortality were noted between HS group and NS group animals,and the survival rate was increased by administration of a combination of L-carnitine and mildronate(Fig.1). Results from previous studies have shown that there can be several causes of death of DS rats fed on high salt diet at the age of fifteen weeks[35].Although we did not observe differences between the functional pa-rameters of the heart and lung to body weight indexes of NS group animals and rats receiving a high salt diet (Tabs.1and2),it has been shown that DS rats of that age can die of heart failure[35],while other studies have shown that heart failure develops with further feeding with high salt diet[18,20].In addition,data from previous studies have shown that DS rats receiv-ing a high salt diet have a high incidence of lethal strokes[27,35].Moreover,DS rats fed with a high salt diet develop marked left ventricular hypertrophy [50]that could also be a cause of myocardial infarc-tion and lethal ventricular arrhythmias[28].Kamei et al.[19]showed that DS rats have increased suscepti-bility to ventricular tachycardia that supports the the-ory about lethal ventricular arrhythmias in DS rats. Although we do not have experimental data in this ex-perimental model on the effects of the tested sub-stances on cardiac arrhythmias and the development of stroke,there is experimental evidence that mildronate and L-carnitine can exert some anti-arrhythmic[31, 47]and cardioprotective[2,10,41]properties.It is therefore tempting to speculate that the beneficial ef-fect of the combination of L-carnitine and mildronate on mortality in DS rats could be linked to their cardio-protective and anti-arrhythmic activities.Analysis of heart rates among the experimental groups revealed that HS group rats on the eighth week of treatment had developed an elevated heart rate compared with NS group rats(Fig.2B).Previously,it has been shown that resting heart rate is an independ-ent predictor of cardiovascular morbidity and mortal-ity,irrespective of the presence of co-morbidities [33].This finding was in compliance with our results,L-carnitine and mildronate is protective in hypertensionReinis Vilskersts et al.Tab.3.Concentrations of L-carnitine,g-butyrobetaine and mildronate in the plasmaNS group HS group HS+M100group HS+C100group HS+CM group Carnitine(µM)55±6*.#23±3#5±1*56±4*.#36±2*.#GBB(µM) 1.5±0.1# 1.5±0.1#8.6±1.0* 2.2±0.4*.#18.1±3.4*.# Mildronate(µM)––98±12–29±3#Concentrations of L-carnitine,g-butyrobetaine and mildronate in plasma were measured by UPLC/MS/MS on the eighth week of the experi-ment.Values are expressed as the mean±SEM of at least three animals.*p<0.05vs.HS group; p<0.05vs.HS+M100groupbecause the highest heart rate was found in HS group animals,as was the highest mortality(Fig.1).Moreo-ver,the administration of a combination of mildronate and L-carnitine decreased heart rate and significantly reduced mortality.Although mildronate treatment for 8weeks also decreased heart rate,the protective effect against mortality was not so pronounced(p<0.06vs. HS group)as in the combination group.These find-ings could also indicate that a reduction of mortality in the HS+CM group is related to the effects of the tested compounds on the heart.Similar to resting heart rate,endothelial dysfunc-tion has been shown to be associated with the occur-rence of cardiovascular events[34]and hypertension-induced organ damage[49].In our study,the effects of treatment with L-carnitine,mildronate or their combination on the development of endothelial dys-function were assessed in isolated aortic rings.The angioprotective effects of L-carnitine in hypertension-induced endothelial dysfunction have been already documented[5,9].In addition,previous studies have also shown the beneficial effects of mildronate treat-ment in diseases involving endothelial dysfunction like atherosclerosis[48].Surprisingly,we did not ob-serve any improvement of endothelial function in aor-tic rings of HS+C100or HS+M100group animals compared to aortic rings of HS group animals(Fig.3). This could be explained by the two times lower dose of L-carnitine used in our experiment compared to other studies that have shown protective effects of L-carnitine on the development of endothelial dysfunc-tion[5,9].While mildronate or L-carnitine treatment had no impact on endothelial dysfunction,administra-tion of their combination attenuated the development of endothelial dysfunction(Fig.3),but similar to the administration of mildronate or L-carnitine alone,did not reduced blood pressure(Fig.2A).Although re-sults from a recent study demonstrated a positive rela-tionship between a reduction in blood pressure and an improvement in arterial endothelium-dependent re-laxation[6],it has been suggested that the mechanism of attenuation of the development of endothelial dys-function was based on direct influence on endothe-lium and vascular tissues,and not on the hypotensive activity[40,46].Our results support this theory,as endothelial function was improved without significant effect on arterial blood pressure.Data from previous studies have shown beneficial effects of the separate administration of L-carnitine or mildronate in the case of different diseases and pa-thologies,e.g.,type2diabetes mellitus[26,30]and cardiovascular diseases[2,10,24].For the first time, our study demonstrates that the simultaneous admini-stration of L-carnitine and mildronate,an inhibitor of carnitine biosynthesis and reabsorption,shows phar-macological effects better than when the two agents are administered separately.Thus,our study showed that L-carnitine and mildronate could share some common properties that are beneficial in the preven-tion of hypertension-induced complications.Although there is not much experimental evidence about protec-tive effects of GBB,our previous study showed that an increase of GBB concentration induced by admini-stration of mildronate inversely correlated with the size of infarction in experimental heart infarction model in rats in vitro[24].Moreover,it has been shown that GBB protects isolated heart from hydro-gen peroxide induced damage[1].Thus,we can hy-pothesize that beneficial effects of administration of combination of mildronate and L-carnitine could be due to increased GBB concentration,but further ex-periments are required to confirm this hypothesis.In summary,our study demonstrated that treatment with a combination of L-carnitine and mildronate is protective in hypertension-related complications. 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