探讨胆碱对H2O2诱导的心肌细胞凋亡的保护作用

左卡尼汀对过氧化氢诱导的心肌细胞凋亡的抑制作用梁春光;戴红良;黄雷;王东海;王洪新【期刊名称】《中国药理学与毒理学杂志》【年(卷),期】2012(26)5【摘要】目的探讨左旋尼汀对心肌细胞凋亡的抑制作用及其可能的作用机制.方法采用培养的新生乳大鼠心肌细胞制备H2O2 200 μmol· L-1氧化损伤模型.实验分为正常对照组、H2O2 200 μmol·L -1模型组、左卡尼汀(0.3,0.6及1.2 mmol· L-1)组、左卡尼汀1.2 mmol·L -1+ H2O2 200 μmol·L-1组.左卡尼汀0.3,0.6和1.2 mmol· L-作用1h后加入H2O2 200 μmol·L-1培养12 h,MTT法测定心肌细胞存活率,流式细胞仪测定心肌细胞的凋亡率,Western印迹法测定胱天蛋白酶3表达；用试剂盒方法检测过氧化物歧化酶(SOD)活性和丙二醛(MDA)水平.左卡尼汀1.2 mmol· L-1作用5 min后加入H2O2 200μmol·L-1,采用Till 阳离子测定系统监测5 min的心肌细胞[Ca2+]i瞬间变化.结果H2O2200 μmol·L-1作用于心肌细胞12 h能引起心肌细胞凋亡.左卡尼汀0.3,0.6和1.2 mmol· L-1能够不同程度的逆转由H2O2所致的损伤,使SOD活性明显增加,MDA水平明显降低(P＜0.01).左卡尼汀1.2 mmol· L-1作用最强,能够明显降低心肌细胞胱天蛋白酶3表达(P＜0.01),明显降低H2O2引起的[Ca2+]i瞬间变化升高(P＜0.01),但对于H2O2引起无钙血清培养的心肌细胞静息钙升高无影响.结论左卡尼汀能够抑制由H2O2引起的心肌细胞凋亡,该抑制作用可能与其保护细胞膜和减轻钙通道的损害、纠正[Ca2+]i瞬变失调及其抗氧化作用有关.%OBJECTIVE To investigate the effect of L-carnitine on H2O2-induced apoptosis in cultured neonatal rat cardiomyocytes and its possible mechanisms.METHODS H2O2 200 μmol· L-1 was used to set upan oxidative stress induced injury model in neonatal rat cardiomyocytes.There were normal control,H2O2 200 μmol·L-1 model,L-carnitine (0.3,0.6 and 1.2 mmol· L-1 ),L-carnitine ( 1.2 mmol· L-1 ) + H2O2 ( 200 μmol· L-1 )groups in the experiment.The cells were pretreated with L-carnitine 0.3,0.6 and 1.2 m mol· L- 1 for 1 h before H2O2 200 μmol·L-1 was added for 12 h.Cell viability was measured by MTT assay and apoptosis was determined by flow cytometry.The expression of caspase 3 was determined by Western blotting.Superoxide dismutase (SOD) and malondialdehyde (MDA) were measured according to the instructions of commercial kits. L-Carnitine 1.2 mmol· L- 1 was added 5 min prior to H2O2 200 μ mol· L- 1.H2O2 was perfused for 5 min to measure [Ca2 +] i transient by Till image system by cell loading Fura-2/AM.RESULTS Application of H2O2 200 μ mol ·L-1 to myocytes for 12 h could cause cardiomyocyte apoptosis.L-Carnitine 0.3,0.6 and 1.2 mmol· L-1 significantly reversed the lesion of primary cultured cardiomyocytes treated with H2O2 200 μmol· L-1,increased SOD activity (P ＜0.01 ),and decreased MDA content( P ＜0.01 ).L-Carnitine 1.2 mmol·L-1 was the most powerful in that it decreased caspase 3 expression(P ＜0.01 ) and the adverse effect ofH2O2 on the [Ca2+]i transient was abolished by L-carnitine pretreatment.L-Carnitine had no effect on the elevation of the resting [Ca2+] i evoked by H2O2 in Ca2+ -free medium. CONCLUSION L-Carnitine can produce an inhibitory effect on cardiomyocyte apoptosis induced by H2O2,which may be mediated by the protective effect on the cellmembrane and the damage to voltagedependent Ca2+ channel as well as its inhibition of lipid peroxidation.【总页数】8页(P602-609)【作者】梁春光;戴红良;黄雷;王东海;王洪新【作者单位】辽宁医学院分子生物学与新药开发重点实验室,辽宁锦州121001;辽宁医学院分子生物学与新药开发重点实验室,辽宁锦州121001;锦州市食品药品检验所,辽宁锦州121000;大同煤总医院,山西大同037003;辽宁医学院分子生物学与新药开发重点实验室,辽宁锦州121001【正文语种】中文【中图分类】R975;R963【相关文献】1.C-sis基因对过氧化氢诱导大鼠肝细胞凋亡的抑制作用 [J], 张弘英;胡树根;元文峰;丁浩2.吡格列酮对过氧化氢诱导的乳鼠心肌细胞凋亡的抑制作用 [J], 王东海;杨育红;戴红良;吴国强3.刺五加叶皂苷B对过氧化氢诱导的心肌细胞凋亡的抑制作用 [J], 梁启明;曲绍春;于晓风;徐华丽;张四维;邹敬涛;常文光;睢大筼4.银杏酮酯对过氧化氢诱导大鼠乳鼠心肌细胞凋亡的抑制作用 [J], 金玉子;杨育红;王洪新;李香华5.家蚕Bm30Kc6蛋白对过氧化氢诱导的家蚕细胞凋亡的抑制作用 [J], 应慧慧;余海鹏;寿鑫;全滟平;张耀洲;童富淡;于威因版权原因，仅展示原文概要，查看原文内容请购买。

金丝桃苷对过氧化氢损伤心肌细胞的保护作用研究梁凯;孔晓龙;覃振明;莫凤珍;汪永玲;黄绍德;郭梅红【摘要】目的研究金丝桃苷对过氧化氢(H2O2)损伤心肌细胞的保护作用.方法以原代培养的乳鼠心肌细胞建立H2O2损伤模型.实验分为7组:正常对照组、模型对照组、二甲基亚砜组(1%二甲基亚砜)、维拉帕米组(25 μmol/L)、金丝桃低剂量组(0.6 μmol/L)、金丝桃中剂量组(6 μmol/L)、金丝桃高剂量组(60 μmol/L).测定各组心肌细胞一氧化氮合酶(NOS)、Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶、肌酸激酶同工酶MB(CK-MB)活性变化及心肌细胞肌钙蛋白(cTnI)的含量.结果金丝桃苷中、高剂量组结构型一氧化氮合酶(cNOS)、总一氧化氮合酶(tNOS)、Na+-K+-ATP酶、Ca2+-Mg2+-ATP酶活性较模型对照组显著增加(P＜0.01),诱导型一氧化氮合酶(iNOS)活性较模型对照组降低(P＜0.05);金丝桃苷低剂量组、中剂量组、高剂量组上清液的CK-MB、cTnI含量较模型对照组低(P＜0.01).结论金丝桃苷具有抗心肌细胞过氧化氢损伤作用,这可能与其提高心肌细胞膜的Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶活性,并调节不同亚型NOS活性有关.%Objective To study the protective effects of hyperin( Hyp ) on the hydrogen peroxide-injured ( H2O2-injured ) myocardial cells. Methods A H2O2-injured model of the primary cultrued neonatal rat cell was established. There were seven experimental groups:normal control group,model control group,DMSO group( 1% DMSO ), Verapamil group( 25 μmol/L ),low-,medium- and high-dosage Hyp groups( 0. 6 ,6 ,60 μmol/L ). The activi ties of NOS, Na +-K + -ATPase,Ca +-Mg + -ATPase, creatine kinase isoenzyme MB( CK-MB ) and content of myocardial troponin I ( cTnl ) in myocardial cells of each group were detected. Results Compared with model control group,the activitiesof cNOS.tNOS, Na+-K +-ATPase, Ca2+-Mg2+-ATPase increased significantly ( P < 0. 01 ) and the activity of iNOS decreased in medium- and high dose Hyp groups( P <0.05 ). The contents of CK-MB,cTnl from the supernatant in low-, medium-, high-dose groups were lower than those of model control group( P <0. 01 ). Conclusion Hyp has protective effect on the H2O2-injured myocardial cells,its mechanism may be related to increasing the activities of Na+-K+-ATPase, Ca + -Mg +-ATPase and regulating the activities of different subtypes of NOS.【期刊名称】《广西医学》【年(卷),期】2012(034)012【总页数】4页(P1600-1603)【关键词】金丝桃苷;心肌细胞;过氧化氢损伤;心肌保护【作者】梁凯;孔晓龙;覃振明;莫凤珍;汪永玲;黄绍德;郭梅红【作者单位】广西医科大学药学院药理学教研室,南宁市,530021;广西医科大学药学院药理学教研室,南宁市,530021;广西医科大学药学院药理学教研室,南宁市,530021;广西医科大学药学院药理学教研室,南宁市,530021;广西医科大学药学院药理学教研室,南宁市,530021;广西医科大学药学院药理学教研室,南宁市,530021;广西医科大学药学院药理学教研室,南宁市,530021【正文语种】中文【中图分类】R542.2;R96金丝桃苷(hyperoside，Hyp)，又名槲皮素-3-O-β-D-吡喃半乳糖苷，属于黄酮醇苷类化合物，具有多种生理活性，可以抗炎、解痉、利尿、止咳、降压、降低胆固醇、蛋白同化等作用［1］。

260新医学综述2024年4月第55卷第4期低温诱导的RBM3对各器官缺血再灌注损伤研究进展孙路轩 党晓平 孙子健【摘要】缺血再灌注损伤（IRI）是一种复杂的血流动力学紊乱状态，致死率极高，且目前的治疗方法相对有限。

亚低温治疗是临床上公认的一种缓解缺血、缺氧损伤的治疗方式，尤其在脑保护中的研究较多。

多项研究表明，RNA 结合基序蛋白3（RBM3）作为一种冷应激蛋白，主要在低温诱导下产生，可促进翻译，减轻氧化应激、降低细胞凋亡率。

因此，诱导RBM3可能代表一种治疗IRI的新策略，代替亚低温治疗，减轻低温对机体的不良影响。

基于这一观点，文章对RBM3蛋白功能的最新发现进行综述，重点关注RBM3对各器官IRI相关疾病的保护作用以及未来前景，为相关研究提供新思路。

【关键词】低温；RNA结合基序蛋白3；脑缺血再灌注损伤；心缺血再灌注损伤；肝缺血再灌注损伤； 肾缺血再灌注损伤Research progress in hypothermia-induced RBM3 in ischemia-reperfusion injury of various organs Sun Luxuan△， Dang Xiaoping， Sun Zijian. △The Second Clinical Medical College of Xi’an Medical College， Xi’an 710038， ChinaCorresponding author: Dang Xiaoping， E-mail:***************【Abstract】 Ischemia-reperfusion injury （IRI） is a complex hemodynamic disorder with high mortality rates and limited treatment options. Mild hypothermia is a widely-accepted treatment method for alleviating ischemic and hypoxic injury， especially in the study of brain protection. Many studies have shown that RNA-binding motif protein 3 （RBM3）， as a cold stress protein， is mainly produced under low-temperature induction， which can promote translation， alleviate oxidative stress， and reduce cell mortality. Therefore， inducing RBM3 may represent a new strategy for treating IRI， replacing mild hypothermia and mitigating the side e ff ects of hypothermia on the body. In this study， the latest fi ndings on the function of RBM3 protein were summarized， highlighting the protective role and future prospect of RBM3 in IRI-related diseases of various organs.【Key words】 Hypothermia； RNA-binding motif protein 3； Brain ischemia-reperfusion injury；Heart ischemia-reperfusion injury； Liver ischemia-reperfusion injury； Renal ischemia-reperfusion injury缺血再灌注损伤（IRI）是指组织缺血一段时间后血流恢复时，组织损伤进一步加重、器官功能进一步恶化的综合征［1-2］。

内质网应激介导过氧化氢诱导的心肌细胞凋亡孔曼;陈娟;周洁;朱文德;万丽敏;熊宇芳;李子希【摘要】Objective To investigate relationship between the apoptosis of cardiomyocyte induced by oxidative stress and the endoplasmic reticulum stress(ERS). Methods The cultured H9C2 cells were divided into 2 groups:low concentrationC 100 jLimol/L H2O2)group and high concentrationC500 jumol/L H2O2)group at different time points. The apoptosis rate was detected by flow cytometry and the typical cardiomyocyte apoptosis was observed by hematoxylin-eosinC HE) staining. The expression of ERS marker proteins p-PERK and CHOP was detected by using Western blot. The chemical chaperone 4-phenylbutyric acid (PBA)was used to inhibit ERS in myocardial cells and the expression of p-JNK, p-PERK and CHOP was detected by using Western blot. The activities of Caspase-12 and Caspase-3 were measured by using flow cytometry. Results ? The highest rate of apoptosis was observed in the myocardial cells treated with H2O2 (100 jumol/Dfor 8 h. (2) The expression of ERS marker proteins p-PERK and CHOP was significantly higher in the H2O2 groups. (3)PBA could effectively inhibit H2O2-induced apoptosis of cardiomyocytes,significantly down-regulate the expression of ERS marker proteins p-PERK and CHOP(P<0. 01) ,and significantly reduce the activities of Caspase-3 and Caspase-12 (P<0. 01). Conclusion The low concentration of H2O2 (100 jLtmol/L)significantly induced the apoptosis of myocardial cells via triggering ERS regulation mechanism and high concentration ofH2 O2 (500 jLimol/Dresulted in myocardial necrosis. Our results could provide new clues for prevention and treatment of cardiovascular diseases.%目的探讨氧化应激诱导心肌细胞凋亡是否与内质网应激(endoplasmic reticulum stress,ERS)的调控机制有关.方法给予乳鼠心肌细胞高低两种浓度(500、100μmol/L)和不同时间(0、4、8、12、24 h)过氧化氢(H2O2)刺激.采用流式细胞仪检测各组心肌细胞的凋亡率;苏木精-伊红染色法(HE)观察心肌细胞凋亡的典型形态;通过Western blot检测ERS标志蛋白p-PERK 和CHOP的表达变化.进一步采用ERS抑制剂化学伴侣PBA(4-phenylbutyric acid)抑制心肌细胞ERS,Westernblot检测p-JNK、p-PERK 和CHOP蛋白的表达变化;采用流式细胞仪检测Caspase-12 和Caspase-3的活性.结果①低浓度H2O2可显著诱导心肌细胞凋亡,高浓度H2O2则导致心肌细胞发生坏死;100 μmol/L H2O2刺激心肌细胞8 h时其凋亡率最高.②心肌细胞发生凋亡时ERS标志蛋白p-PERK 和CHOP表达显著增高.③ERS抑制剂PBA预处理心肌细胞,可有效抑制H2O2诱导的p-PERK、CHOP表达及Caspase-3、Caspase-12活性的上调,与单纯H2O2处理组相比差异有统计学意义(P＜0.01).结论低浓度H2O2可通过促发ERS整合调控机制介导心肌细胞凋亡.这一结果将从ERS整合调控细胞应激的角度为心血管疾病的防治提供新思路.【期刊名称】《华中科技大学学报（医学版）》【年(卷),期】2012(041)003【总页数】5页(P253-257)【关键词】内质网应激;氧化应激;细胞凋亡;心肌细胞【作者】孔曼;陈娟;周洁;朱文德;万丽敏;熊宇芳;李子希【作者单位】华中科技大学同济医学院生物化学与分子生物学系,武汉,430030;武汉市中心医院检验科,武汉,430014;华中科技大学同济医学院生物化学与分子生物学系,武汉,430030;华中科技大学同济医学院生物化学与分子生物学系,武汉,430030;华中科技大学同济医学院生物化学与分子生物学系,武汉,430030;华中科技大学同济医学院生物化学与分子生物学系,武汉,430030;华中科技大学同济医学院生物化学与分子生物学系,武汉,430030;华中科技大学同济医学院附属中西医结合医院检验科,武汉,430070【正文语种】中文【中图分类】R349.5研究表明，心肌细胞凋亡可参与多种心血管疾病的发生过程，如冠心病、心绞痛、心力衰竭、心肌炎及心脏自身免疫性疾病等［1］。