肝肾阴虚证大鼠下丘脑—垂体—甲状腺轴的变化及中药对其调节作用

NEURAL REGENERATION RESEARCH Volume 7, Issue 21, July 2012Cite this article as: Neural Regen Res. 2012;7(21):1611-1617.1611Siyun Niu ☆, M.D., Associate professor, Master’ssupervisor, School of Basic Medical Science, Hebei University, Baoding 071000, Hebei Province, ChinaSiyun Niu and Suru Kou contributed equally to this study.Corresponding author: Siyun Niu, School of Basic Medical Science, Hebei University, Baoding 071000, Hebei Province, China nsy1688@Received: 2012-02-15 Accepted: 2012-05-13 (N20111107001/WJ)Niu SY , Kou SR, Zhou XC, Ding L. Heshouwu decoction, a Chinese herb for tonifying kidney, ameliorates hypothalamic-pituitary- testicular axis secretion in aging rats. Neural Regen Res. 2012;7(21):1611-1617.doi:10.3969/j.issn.1673-5374.2012.21.002Heshouwu decoction, a Chinese herb for tonifying kidney, ameliorates hypothalamic-pituitary- testicular axis secretion in aging rats****☆Siyun Niu 1, Suru Kou 1, Xiaochun Zhou 2, Liang Ding 11School of Basic Medical Science, Hebei University, Baoding 071000, Hebei Province, China2Institute of Basic Medical Sciences, Chengde Medical College, Chengde 067000, Hebei Province, ChinaAbstractAn increasing amount of evidence demonstrates the anti-aging effect of Heshouwu in pill form. In this study, a subacute aging rat model was established by continuous intraperitoneal injection of D-galactose and treated with Heshouwu decoction (a Chinese herb for tonifying the kidney,comprising Heshouwu pill, Herba Epimedii, Radix Salviae Miltiorrhiae, and Poria). Heshouwu pill treated rats were the positive control group. Radioimmunoassay, immunohistochemical staining, and western blot assay showed hypothalamic gonadotropin-releasing hormone, hypothalamic substance P , and serum gonadotropin levels to be significantly increased in the model rats; the concentrations of hypothalamic β-endorphin, and serum levels of insulin-like growth factor 1 and testosterone were significantly decreased. 17β- and 3β-hydroxysteroid dehydrogenase expression in testicular tissue was also decreased. Intragastric administration of Heshouwu decoction at high (9.6 g/mL/100 g), medium (4.8 g/mL/100 g), and low (2.4 g/mL/100 g) doses, Heshouwu decoction pretreatment at a medium dose (4.8 g/mL/100 g), and Heshouwu pill (2.06 g/mL/100 g) significantly reversed these changes. Heshouwu decoction pretreatment and high-dose Heshouwu decoction had the greatest anti-aging effects. These experimental findings indicate that Heshouwu decoction can improve hypothalamic-pituitary-testicular axis secretion in a subacute aging rat model, and prevent and delay gonadal axis aging, with an effect superior to that of Heshouwu pill.Key WordsHeshouwu decoction; pituitary gonadal axis; aging; gonadotropin-releasing hormone; gonadotropin; hydroxysteroid dehydrogenase; hypothalamic-pituitary-testicular axis; neural regenerationResearch Highlights(1) Gonadotropin-releasing hormone, substance P , and gonadotropic hormone are significantly increased in the hypothalamic-pituitary-testicular axis of aging rats, whereas β-endorphin,insulin-like growth factor-1, testosterone concentration, 17β-hydroxysteroid dehydrogenase and 3β-hydroxysteroid dehydrogenase are significantly decreased. (2) Heshouwu decoction can reverse these phenomena and its effect is superior to that of Heshouwu pill.AbbreviationsGnRH, gonadotropin-releasing hormone; LH, luteinizing hormone; FSH, follicle-stimulating hormone; IGF-1, insulin-like growth factor 1; HSD, hydroxysteroid dehydrogenase1612INTRODUCTIONAging is an irreversible, natural phenomenon, andgonadal hormones may directly or indirectly regulate the aging process [1]. This study aimed to improve gonadal hormone levels, which are closely related to human aging. An increasing amount of evidence has demonstrated the anti-aging effect of Heshouwudecoction, and the underlying mechanism may depend on the following factors [2-3]: (1) improved antioxidantcapacity and regulation of blood lipids; and (2) influences on p53/pRb related protein expression in the aging pathway in testicular cells. However, the influence of Heshouwu decoction on hypothalamic-pituitary-testicular axis secretion and on the activity of the key enzymes in testosterone biosynthesis remains unclear.In the present study, Heshouwu decoction pretreatment and treatment was given to a D-galactose-induced rat aging model and hypothalamic-pituitary-testicular axis secretion was investigated in an attempt to understand the regulatory effect on gonadal axis secretion.RESULTSQuantitative analysis of experimental animalsEighty Sprague-Dawley rats, after 1 week of adaptive feeding, were divided randomly into seven groups. Normal group (n = 10) rats were injectedintraperitoneally with normal saline. For the model group (n = 10), subacute aging was induced by intraperitoneal injection of D-galactose. In themedium-dose Heshouwu decoction pretreatment group (n = 12), rats were pretreated with 4.8 g/mL/100 gHeshouwu decoction to observe the protective effect of this drug against D-galactose-induced subacute aging. In the Heshouwu decoction treatment groups (n = 12), following intraperitoneal injection of D-galactose, rats were injected with Heshouwu decoction at a high(9.6 g/mL/100 g), medium (4.8 g/mL/100 g), or low (2.4 g/mL/100 g) dose, or given Heshouwu in pill form (2.06 g/mL/100 g) suspension, to observe thetherapeutic effects on D-galactose-induced subacute aging. In the natural recovery group, model rats were allowed to recover spontaneously, to determine whether recovery from D-galactose-induced subacute aging can occur naturally over time. All 80 rats were involved in the final analysis.17β-hydroxysteroid dehydrogenase (HSD) and 3β-HSD expression in rat testicular tissueImmunohistochemical staining showed that 17β-HSD and 3β-HSD were expressed in a dispersed pattern inthe cytoplasm of rat Leydig cells. Semi-quantitative analysis by western blot assay demonstrated that17β-HSD and 3β-HSD protein expression in the model rats was significantly lower than that in the normal group (P < 0.01). Expression was increased to varying degrees in the Heshouwu decoction pretreatment group, theHeshouwu decoction treatment groups, and the Heshouwu pill group (P < 0.05). Levels in the Heshouwu decoction pretreatment group and the high-dose Heshouwudecoction treatment group were higher than those in the other groups (P < 0.05; Table 1, Figures 1 and 2). These symptoms were apparently reversed after intragastric administration of Heshouwu decoction at high (9.6 g/mL/100 g), medium (4.8 g/mL/100 g), and low (2.4 g/mL/100 g) doses, by Heshouwu decoction pretreatment (4.8 g/mL/100 g), and by Heshouwu pill (2.06 g/mL/100 g).Gonadotropin-releasing hormone (GnRH), substance P and β-endorphin changes in rat hypothalamus Radioimmunoassay showed that hypothalamic GnRH and substance P levels were significantly increased in the model rats compared with the normal group (P < 0.01), whereas β-endorphin was significantly decreased (P < 0.01). High-dose Heshouwu decoction andHeshouwu decoction pretreatment were superior to the other treatments (P < 0.05; Table 2).Serum levels of luteinizing hormone (LH),follicle-stimulating hormone (FSH), testosterone, and insulin-like growth factor 1 (IGF-1)Radioimmunoassay showed that serum LH and FSH levels in the model rats were significantly higher than those in the normal group (P < 0.01), whereastestosterone levels were significantly decreased (P < 0.01).Enzyme-linked immunosorbent assay demonstrated thatthe IGF-1 level in aging rats was significantly lower thanthat in the normal rats (P < 0.01). Serum LH and FSHlevels were decreased to varying degrees in theHeshouwu decoction pretreatment group, the Heshouwudecoction treated groups, and the Heshouwu pill treatedgroup, but IGF-1 and testosterone levels were increased(P < 0.01). Heshouwu decoction pretreatment and high-and medium-dose Heshouwu decoction were superior tolow-dose Heshouwu decoction and Heshouwu pill (P <0.05; Table 3).Figure 1 17β-HSD and 3β-HSD expression intesticular tissue (immunohistochemical staining,substance P method, optical microscope, × 400).17β-HSD (A–D) and 3β-HSD (E-H) were expressedpredominantly in the cytoplasm of Leydig cells.In the model group (A, E), 17β-HSD and 3β-HSDpositive products were stained light yellow.In the normal group (B, F), the Heshouwu decoctionpretreatment group (C, G), and the high-doseHeshouwu decoction group (D, H), the positiveproducts were brown, and their levels were greaterthan in the model group. (I) Negative control group.HSD: Hydroxysteroid dehydrogenase.A B CD E FG H IFigure 2 17β-HSD and 3β-HSD protein expression intesticular tissue.1: Model group; 2: normal group; 3: Heshouwu decoctionpretreatment group; 4: high-dose Heshouwu decoctiongroup; 5: medium-dose Heshouwu decoction group; 6:low-dose Heshouwu decoction group; 7: Heshouwu pillgroup; 8: spontaneous recovery group.17β-HSD and 3β-HSD protein expression was greatest inthe Heshouwu decoction pretreatment and high-doseHeshouwu decoction groups.HSD: Hydroxysteroid dehydrogenase.17β-HSD3β-HSDβ-actin1234567834 kDa43 kDa43 kDa1613DISCUSSIONThe D-galactose-induced subacute aging model is based on Senescence Metabolism Theory. The aging phenomena induced by D-galactose reflect the natural state of senescence within a short period of time, so this model is commonly used[4-5]. The drug is administered via intraperitoneal injection at a dose of 40-500 mg/kg per day for 20-60 days[6]. This study used intraperitoneal injection of D-galactose to produce a subacute aging model in the rat.Increasing evidence indicates the anti-aging effects of various single herbs and herbal compounds. Wanget al [7] demonstrated that saponins in Radix Ginseng, American Ginseng, and Radix Notoginseng can scavenge free radicals. Barbarum polysaccharide also exhibits an antioxidative capacity, with effects such as improving superoxide dismutase activity in the serum, heart, liver, and brain tissue in a D-galactose-induced aging mouse model, reducing malondialdehyde, and increasing serum and cardiac telomerase activity[8]. Weng et al [9] studied the influence of Ganodermasides A and B (two monomer compounds) in Ganoderma on the replication and survival of Saccharomyces cerevisiae, and found an anti-aging effect. Many Chinese herbal compounds, such as Apozem of Tremella and Wolfberry Fruit, Yangzhen Oral Liquid, and Yishuotiaozi Tablet, increase the activity of antioxidant enzymes, reduce malondialdehyde accumulation, and improve immunity. Vitality Reinforcing Prescription downregulates expression of the tumor suppressor gene p53 in spleen and liver cells of aging male mice at the transcription and translation levels. Jinguishenqi Pills are well known as antagonists of the DNA damage caused by cyclophosphamide. Liuwei Dihuang decoction can prolong the survival of Drosophila and increase telomerase activity in the brain and gonadal tissue of aging mice[10]. However, there is little evidence available on pituitary-testicular axis secretion, and the present study is the first demonstration of the action of Heshouwu decoction on pituitary-testicular axis secretion.Heshouwu decoction can regulate serum antioxidant capacity and lipid metabolic disorders[2]; through regulation of Rb/p16 and p53/p19/p21, this decoction also relieves cell conduction blockade, promotes the proliferation of ovarian and testicular cells, and inhibits apoptosis[3, 11-13]. Less attention had been paid to the regulation by Heshouwu decoction of testosterone. Our study showed that serum testosterone levels were significantly lower in aging rats than in the normal group, and Heshouwu decoction apparently improved serum testosterone levels in a dose-dependent manner. Pretreatment with Heshouwu decoction achieved optimal anti-aging effects.Testosterone synthesis and secretion are modulated by hypothalamic GnRH, peptide neurotransmitters, gonadotropins, and growth hormones secreted from the pituitary gland. Pituitary endocrine dysfunction is closely related to hypothalamic regulatory function[14], and the pulse and wave coordination of pituitary gonadotropic hormone and sex hormone secretion is decreased or absent in older males and females[15-16]. The hypothalamus-generated neuropeptides substance P and β-endorphin are also involved in GnRH synthesis and secretion[17]. By a feedback mechanism, testosterone can inhibit hypothalamic GnRH, pituitary FSH, LH, and IGF-1 levels. The role of Heshouwu decoction in this process has not yet been elucidated. In the present study, increased levels of hypothalamic substance P and GnRH, significantly decreasedβ-endorphin, and higher levels of serum FSH, LH, and IGF-1 were observed in aging rats and normal rats, and reversed by Heshouwu decoction. There is evidence that kidney-tonifying Chinese herbs regulate testosterone through hypothalamic GnRH secretion and release of the neurotransmitters β-endorphin and substance P, thereby affecting the secretion of FSH, LH, and IGF-1, or through the regulation of testosterone secretion by Leydig cells, providing negative feedback regulation of hypothalamic and pituitary function when gonadal axis secretion becomes abnormal as aging proceeds. Our findings support the conclusion that kidney-tonifying Chinese herbs regulate testosterone levels through the hypothalamic-pituitary axis.Leydig cell-secreted steroid dehydrogenases play an important regulatory role in testosterone synthesis and secretion. Various factors contribute to the influence on testosterone synthesis of 3β-HSD and 17β-HSD activity[18-20], but the role of Heshouwu decoction on3β-HSD and 17β-HSD synthesis and secretion in Leydig cells is unclear. This study observed 3β-HSD and 17β-HSD expression in testis tissue from aging rats and explored the regulatory effect of Heshouwu decoction.We found that 3β-HSD and 17β-HSD protein expression was significantly lower in aging rats than in normal rats, and Heshouwu decoction upregulated their expression in a dose-dependent manner. These experimental findings indicate that Heshouwu decoction can increase testosterone secretion in Leydig cells and regulate 3β-HSD and 17β-HSD levels. In summary, Heshouwu decoction can ameliorate hypothalamic-pituitary-testicular axis secretion in a subacute aging rat model, and prevent and delay gonadal axis aging, with an effect greater than that of Heshouwu pills.1614MATERIALS AND METHODSDesignA randomized, controlled animal experiment.Time and settingExperiments were performed from November 2009 to September 2010 at the Cell Biology Laboratory, School of Basic Medical Science, Hebei University, China.MaterialsAnimalsNinety clean, healthy male Sprague-Dawley rats, weighing 180-220 g, were provided by the Experimental Animal Center of Hebei Medical University, China (license No. 1005050).All experimental use of animals complied with the Guidance Suggestions for the Care and Use of Laboratory Animals, issued by the Ministry of Science and Technology of China[21].Chinese medicine compoundsHeshouwu decoction prescription: The decoction comprised Radix Polygoni Multiflori, Herba Cistanches, Achyranthes bidentata Blume, epimedium, Salvia miltiorrhiza, and tuckahoe (Traditional Chinese Medicine Hospital of Hebei Province, China) according to the ratio 3:2:3:2:5:3, immersed in eight times the volume of distilled water for 1 hour. The herbs were then decocted twice in simmer water, for 30 minutes each time. The decoction was condensed to high-, medium-, and low-dose liquids containing 2.4, 4.8, and 9.6 g/mL crude drug, respectively. All decoctions were stored at 4°C and rewarmed to 25-30°C before administration.Heshouwu pill prescription: The pill comprised Radix Polygoni Multiflori, Herba Cistanches, and Achyranthes bidentata Blume (Traditional Chinese Medicine Hospital of Hebei Province, China) according to the ratio 3:2:3, immersed in eight times the volume of distilled water for 1 hour. The herbs were then decocted twice in simmer water, for 30 minutes each time. The decoction was condensed to liquid containing 2.06 g/mL crude drug, which was stored at 4°C and re-warmed to 25-30°C before administration.Drug doses: All drug doses were calculated based on the adult dose[22]. Heshouwu decoction containing 2.4 g/mL crude drug was equivalent to a low adult dose, the medium dose was twice the low adult dose, and the high dose was four times the low adult dose. Heshouwu pill (2.06 g/mL) corresponded to the adult dose. Rats were given 1 mL/100 g at 15:00 each day for 60 consecutive days. MethodsEstablishment of subacute aging rat modelEighty 8-week-old, clean grade, healthy maleSprague-Dawley rats, weighing 180-220 g, were used in this study to produce the subacute aging model. Under aseptic conditions, D-galactose (Beijing Chemical Reagent Company, Beijing, China) was soaked in normal saline to prepare a 6% solution, which was given to the rats via intraperitoneal injection at a dose of 300 mg/kg per day for 60 consecutive days[3-6]. Other groups of rats were injected with normal saline solution for 60 days, once per day.Rat testis tissue and brain tissue samplesRats were killed under 10% chloral hydrate anesthesia, and blood samples of 2 mL were collected using a capillary pipette inserted into the medial orbital venous plexus and centrifuged at 800 ×g for 20 minutes. The supernatant was discarded and stored at -80°C. Following thoracotomy, cannulation was performed from the left ventricle to the ascending aorta for rapid infusion of cold saline (approximately 200 mL), then bilateral testicular tissue and brain tissue were rapidly removed. The left testis and the brain tissue were preserved in liquid nitrogen for 30 minutes and stored at -80°C. The right testis was fixed in paraformaldehyde, embedded in paraffin, and stained immunohistochemically.Radioimmunoassay for serum LH, FSH, and testosterone levelsA radioimmunoassay kit (Beijing Northern Biotechnology Research Institute, Beijing, China) was used to determine the serum levels of LH, FSH, and testosterone in strict accordance with the manufacturer’s instructions. Levels were calculated using a standard curve method.Enzyme-labeled immunosorbent assay for serum IGF-1 levelsAn enzyme-labeled immunosorbent assay kit (Wuhan Boster Biological Engineering Co., Ltd., Wuhan, Hubei Province, China) was employed to measure serum IGF-1 (rabbit anti-IGF-1 monoclonal antibody, goat anti-rabbit IgG; 78-5 000 pg/mL) in strict accordance with the manufacturer’s instructions. Levels were calculated using a standard curve method.Radioimmunoassay for hypothalamic GnRH, substance P, and β-endorphin levelsFive rats in each group were selected for harvesting of brain tissue. The hypothalamus was removed to the rear of the optic chiasma at the ventral side and weighed (approximately 30 mg). The specimens were then homogenized with 400 μL cell lysate, ground in an ice bath, and centrifuged at 4°C; the supernatant was1615discarded. GnRH, substance P, and β-endorphin were measured using a radioimmunoassay kit (PLA Navy Radioimmunoassay Center, China) according to the manufacturer’s instructions. Serum levels were calculated using a standard curve method.Immunohistochemical staining for 17β-HSD and3β-HSD expression in testicular tissueSlices were dewaxed to water and rinsed with PBS (0.01 M, pH 7.4) three times for 5 minutes each time, then incubated with 3% hydrogen peroxide-methanol solution at room temperature for 30 minutes to remove endogenous peroxidase activity. The specimens were incubated with normal goat serum at 37°C for30 minutes, after which the serum was absorbed with filter paper and the specimens were incubated with rabbit anti-17β-HSD (1:100) and anti-3β-HSD monoclonal antibody (1:50; Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 4°C overnight (20 hours), biotin-labeled secondary antibody (goal anti-rabbit IgG; Santa Cruz) at 37°C for 30 minutes and horseradish peroxidase-conjugated streptavidin (Beijing Zhongshan Jinqiao Biological Co., Ltd., Beijing, China) at 37°C for 30 minutes, and subjected to 3,3’-diaminobenzidine coloration (Beijing Zhongshan Jinqiao) under optical microscopy (Leica DM6000M, Wetzlar, Germany). Between each culture step, the specimens were rinsed with PBS three times for 5 minutes each time. After termination of the coloration, the specimens were counterstained with hematoxylin, dehydrated in an alcohol gradient, rendered transparent in xylene, and mounted with neutral gum. For negative controls, PBS was used instead of antibody, with the other steps being the same as those described above. The appearance of brownish yellow particles or fine particles in a diffuse distribution was considered a positive response.Western blot assay for 17β-HSD and 3β-HSD expression in testicular tissueTotal protein was extracted with a cell lysis kit and the total protein content of the specimens was determined with a bicinchoninic acid quantitative protein detection kit. Four per cent stacking gel and 10% separation gel were prepared, and 60 µg of testicular tissue protein was mixed with sampling buffer solution, boiled in a water bath for 5-10 minutes, and placed in the gel sample holes after natural cooling. Electrophoresis was performed under 80 V on the stacking gel and 100 V on the separation gel. The extracted proteins were transferred to polyvinylidene fluoride membrane using a water-bath electric transfer device at 4°C, at 30 V constant pressure overnight. The membrane was blocked with 5% skimmed milk powder and shaken gently for 1 hour at room temperature. The blocked polyvinylidene fluoride membrane was dried, placed into the hybridization bag, and incubated with rabbitanti-17β-HSD, 3β-HSD, and β-actin monoclonal antibodies (1:200; Santa Cruz) for 2 hours. The membrane was rinsed with TBST membrane three times for 10 minutes each time and oscillated gently, then dried and incubated with horseradish peroxidase-conjugated secondary antibody (goat anti-rabbit IgG antibody,1:2 000; Santa Cruz) at 37°C for 1 hour and rinsed in TBST three times for 10 minutes each time. Following polyvinylidene fluoride membrane coloration, Pierce chemiluminescent substrate solutions A and B were mixed for 15 minutes at room temperature and the scanned image was analyzed with a FUJI Mini-4000 (Fuji, Tokyo, Japan).Statistical analysisMeasurement data were expressed as mean ± SD using SPSS 16.0 software (SPSS, Chicago, IL, USA). Differences between groups were compared withone-way analysis of variance, and paired comparisons were made using the Student-Newman-Keuls test. P values less than 0.05 were considered significant.Funding:This study was supported by the Talent Introduction Fund of Hebei University, No. 2010-183; the Medical Science Special Fund of Hebei University, No. 2012A1005; the Key Project of Hebei Provincial Health Department, No. 20110151; and a grant from Hebei Provincial Administration of Traditional Chinese Medicine, No. 2011104.Author contributions: Siyun Niu had full access to the study concept and design, wrote the manuscript and managed the funds. Suru Kou was responsible for data collection and integration. Xiaochun Zhou performed statistical processing and data analysis. Liang Ding validated the study.Conflicts of interest:None declared.Ethical approval:This pilot was approved by the Experimental Animal Ethics Committee of Hebei University in China.REFERENCES[1] Bartke A. 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补肾中药对下丘脑GnRH、垂体FSH、LH及成骨细胞BGP基因表达的调节作用蔡德培;张炜【期刊名称】《中医杂志》【年(卷),期】2002(43)3【摘要】目的:探讨补肾中药对下丘脑促性腺激素释放激素(GnRH)、腺垂体促性腺激素卵泡刺激素(FSH)、黄体生成素(LH)及成骨细胞骨钙素(BGP)基因表达的调节作用。

方法:采用定量的逆转录-聚合酶链式反应(定量的RT—PCR)方法,观察青春期大白鼠喂饲滋阴泻火或益肾填精中药后,下丘脑GnRH、腺垂体FSH、LH及成骨细胞BGP的信使核糖核酸(mRNA)表达水平的变化。

结果:滋阴泻火药可使下丘脑GnRH、腺垂体FSH、LH及成骨细胞BGP的mRNA表达水平显著下调,而益肾填精药可使下丘脑GnRH、腺垂体FSH、LH及成骨细胞BGP的mRNA表达水平显著上调。

结论:说明补肾中药可在转录水平调节下丘脑GnRH、腺垂体FSH、LH及成骨细胞BGP基因表达,这可能是补肾中药调整性早熟患儿青春发育进程和改善骨骼发育的主要作用机理之一。

【总页数】3页(P221-223)【关键词】基因表达;药物作用;补肾药;下丘脑;垂体系统;补肾中药【作者】蔡德培;张炜【作者单位】上海医科大学儿科医院中西医结合研究室;上海帕赛克生物工程研究所基因组研究室【正文语种】中文【中图分类】R285.5【相关文献】1.大黄不同萃取物对大鼠下丘脑和腺垂体超微结构及其GnRH、GnRH-R和FSHβ表达的影响 [J], 赵盼盼;佟继铭;马淑月;高飞;郭建恩;张树峰2.重组GnRH主动免疫公猪的效果及对垂体GnRH受体、FSHβ和LHβ基因表达的影响 [J], 方富贵;蒲勇;王索路;王琳;章孝荣;陶勇;张运海;刘亚;李运生3.肾阳虚大鼠下丘脑-垂体-性腺轴钙调蛋白的基因表达及补肾中药的调整作用 [J], 蒋淑君;崔存德;许兰芝4.Regulative Actions of the Chinese Drugs for Tonifying the Kidney on Gene Expression of the Hypothalamic GnRH,Pituitary FSH,LH and Osteoblastic BGP [J], 蔡德培;张炜;王友京5.中药对大鼠下丘脑生长抑素及垂体生长激素基因表达与蛋白表达的调节作用 [J], 李嫔;向正华;蔡德培因版权原因，仅展示原文概要，查看原文内容请购买。

补肾疏肝方对神经性厌食大鼠下丘脑-垂体-卵巢轴的调控机制研究胡文晓【摘要】目的探讨补肾疏肝方对神经性厌食(anorexia nervosa,AN)应激模型大鼠下丘脑-垂体-卵巢轴(hypothalamus-pituitary-ovarian axis,HPOA)的调控机制.方法按照隔离、限食、束缚的方法建立AN大鼠应激模型,设立正常对照组(以下简称正常组)、模型组、安慰剂组和补肾疏肝方组(以下简称中药组),测定下丘脑多巴胺(dopamine,DA)、去甲肾上腺素(noradrenaline,NE)和5-羟色胺(5-hydroxytryptamine,5-HT).结果下丘脑DA水平:安慰剂组明显高于正常组(P＜0.05),模型组高于正常组,中药组低于安慰剂组,但无统计学差异.下丘脑NE水平:正常组明显高于模型组和安慰剂组(P＜0.05),中药组高于模型组和安慰剂组,但无统计学差异.下丘脑5-HT水平:各组之间无明显差异.结论补肾疏肝方可以调控AN应激模型大鼠的HPOA功能,可能是通过调控下丘脑DA、NE水平来实现的.【期刊名称】《泰山医学院学报》【年(卷),期】2016(037)001【总页数】2页(P65-66)【关键词】补肾疏肝方;神经性厌食;应激模型;下丘脑-垂体-卵巢轴【作者】胡文晓【作者单位】泰安市妇幼保健院,山东泰安271000【正文语种】中文【中图分类】R741.02神经性厌食(anorexia nervosa, AN)是严重威胁女性青少年身心健康且伴有内分泌异常的心身疾病。

常由于特殊的精神心理变态而导致进食量减少，进而造成严重的营养不良、体重减轻及下丘脑-垂体-卵巢轴(hypothalamus-pituitary-ovarian axis, HPOA)功能紊乱[1-2]。

AN在治疗方面尚无理想的方法，补肾疏肝方临床治疗AN患者有较好的疗效。

本研究应用AN大鼠应激模型，从神经内分泌方面探讨其可能的作用机制。

中药对甲状腺功能异常的治疗作用研究摘要：近年来，甲状腺功能异常问题日益引起医学界的广泛关注。

本文通过深入分析中药在治疗甲状腺功能异常中的作用机制，结合现代药理学研究，探讨了中药治疗的独特优势及潜在应用前景。

本文的研究结果表明，中药多成分、多靶点的特点使其在调节甲状腺功能方面展现出显著疗效，且副作用相对较小。

本文为中药在甲状腺疾病治疗中的应用提供了理论依据，也为未来药物研发指明了方向。

关键词：甲状腺功能异常；中药治疗；药理作用；数据统计分析；研究进展一、引言1.1 研究背景与意义甲状腺功能异常是一种常见的内分泌疾病，包括甲状腺功能亢进和甲状腺功能减退两种情况。

随着现代社会生活节奏的加快和环境污染的加剧，甲状腺疾病的发病率逐年上升，严重影响人们的身体健康和生活质量。

西医治疗虽然效果显著，但长期服用合成甲状腺激素或抗甲状腺药物会带来一系列副作用和并发症。

因此，寻找安全有效的替代疗法成为医学研究的热点之一。

中药作为中国传统医学的重要组成部分，具有悠久的历史和丰富的实践经验。

近年来，越来越多的研究表明，中药在调节甲状腺功能方面具有独特优势。

中药成分复杂，可通过多途径、多靶点发挥作用，既能有效缓解症状，又能从根本上调节机体平衡，减少复发风险。

中药毒副作用较小，更适合长期服用。

因此，深入研究中药对甲状腺功能异常的治疗作用机制，对于提高临床疗效、减少药物副作用具有重要意义。

1.2 研究目的与方法本文旨在系统梳理中药在治疗甲状腺功能异常方面的研究进展，通过分析其作用机制、临床应用及实验研究等方面的内容，揭示中药治疗的优势和潜力。

结合现代科技手段，如高通量测序、网络药理学等技术，深入探索中药治疗甲状腺功能异常的新策略和新方法。

本文采用文献综述、数据分析和案例研究相结合的方法进行。

通过查阅大量国内外相关文献，收集整理关于中药治疗甲状腺功能异常的研究资料；运用数据统计分析方法，对收集到的数据进行整理和归纳；结合具体案例进行分析和讨论，以期得出更加全面、准确的结论。

肾气丸对肾阳虚大鼠下丘脑—垂体—甲状腺轴的调节作用初探金蓉家;杨元宵;邢桂英;陈宜涛;李昌煜【期刊名称】《浙江中医杂志》【年(卷),期】2013(048)005【摘要】目的:探讨肾气丸对肾阳虚证模型大鼠下丘脑—垂体—甲状腺轴的调节作用.方法:将SD大鼠随机分为正常对照组、肾阳虚模型组、肾气丸组、附桂组.采用肌内注射氢化可的松制备肾阳虚模型,造模同时给予治疗药物,并观察大鼠一般状况与体征.连续给药20天后,检测下丘脑TRH、血清TSH、T3、T4的含量.结果:模型组大鼠出现体重增长缓慢、尿量增加、自主活动减少、抓取反应迟钝等表现,下丘脑TRH、血清T3、T4含量下降,血清TSH含量上升；经肾气丸干预后,体征和生化指标均得到改善.结论:肾气丸能改善肾阳虚证引起的下丘脑—垂体—甲状腺轴的功能紊乱.【总页数】2页(P370-371)【作者】金蓉家;杨元宵;邢桂英;陈宜涛;李昌煜【作者单位】浙江中医药大学药学院浙江杭州310053;浙江中医药大学药学院浙江杭州310053;浙江中医药大学药学院浙江杭州310053;浙江中医药大学药学院浙江杭州310053;浙江中医药大学药学院浙江杭州310053【正文语种】中文【相关文献】1.盐巴戟对肾阳虚大鼠下丘脑-垂体-甲状腺轴的调节作用 [J], 翟旭峰;黄玉秋;史辑;贾天柱;娄勇军;董权锋2.右归丸对肾阳虚大鼠下丘脑-垂体-甲状腺轴的动态影响 [J], 陈李圳;王秀凤;马娜;廖成彬;刘基;赖炳荣;黄沐阳;陈树龙3.肾阳虚大鼠下丘脑-垂体-甲状腺轴内分泌及钙调蛋白基因表达与淫羊藿总黄酮的干预 [J], 许兰芝;蒋淑君4.牡蛎肽对肾阳虚大鼠下丘脑-垂体-甲状腺轴调节作用的研究 [J], 李亚;王通;王广飞;王运智;于雁飞;高永林5.金匮肾气丸对“劳倦过度、房室不节”肾阳虚模型小鼠下丘脑-垂体-肾上腺轴功能的影响 [J], 许翠萍;孙静;朱庆均;宋洁;张丹;李震因版权原因，仅展示原文概要，查看原文内容请购买。

肾阳虚大鼠下丘脑-垂体-性腺轴钙调蛋白的基因表达及补肾中药的调整作用蒋淑君;崔存德;许兰芝【期刊名称】《中国组织工程研究》【年(卷),期】2004(008)024【摘要】目的:从分子水平探讨肾阳虚证病理改变及补肾中药淫羊藿总黄酮作用机制,为肾阳虚证康复和开发药物的临床新用途提供理论依据.方法:用大剂量外源糖皮质激素建立肾阳虚动物模型,以实时荧光定量RT-PCR技术,测定各组大鼠下丘脑-垂体-性腺轴钙调蛋白(CaM)mRNA的表达,以及淫羊藿总黄酮对其的影响.结果:肾阳虚大鼠下丘脑、性腺CaM mRNA水平升高,垂体组织CaMmRNA水平没有显著变化,淫羊藿总黄酮能够降低下丘脑、睾丸组织CaM mRNA水平,但对垂体影响不显著.结论:下丘脑中CaM mRNA水平升高与肾阳虚证有关,下丘脑、睾丸组织CaMmRNA水平的降低是淫羊藿总黄酮改善肾阳虚症状的作用机制之一.【总页数】2页(P5056-5057)【作者】蒋淑君;崔存德;许兰芝【作者单位】滨州医学院生理教研室,山东省滨州市,256603;滨州医学院生理教研室,山东省滨州市,256603;潍坊医学院药理教研室,山东省潍坊市,261042【正文语种】中文【中图分类】R932【相关文献】1.淫羊藿总黄酮对肾阳虚大鼠下丘脑-垂体-肾上腺轴钙调蛋白基因表达的影响 [J], 蒋淑君;王桂兰;崔存德;徐红2.赛庚啶对大鼠下丘脑-腺垂体-性腺轴和钙调蛋白基因表达的影响 [J], 胡庆伟;康白;高尔;李广宙;李锋杰3.赛庚啶对大鼠下丘脑—腺垂体—性腺轴和催乳素细胞分泌功能及钙调蛋白基因表达的影响 [J], 胡庆伟;康白;等4.肾阳虚大鼠下丘脑-垂体-甲状腺轴内分泌及钙调蛋白基因表达与淫羊藿总黄酮的干预 [J], 许兰芝;蒋淑君5.奶母果对肾阳虚大鼠下丘脑-垂体-性腺轴中钙调蛋白mRNA表达量的影响 [J], 余世荣因版权原因，仅展示原文概要，查看原文内容请购买。