降糖舒心方对糖尿病心肌内质网应激c-JNK凋亡旁路的影响
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摘要
目的:探讨降糖舒心方(JTSX)抑制SD糖尿病大鼠心肌细胞内质网应激c-JNK凋亡旁路,减轻糖尿病心肌重构的分子机制。方法:采用高脂饲养+链脲佐菌素制备2型糖尿病大鼠模型,成模后随机分为模型组、西药组、JTSX低剂量组(JTSX1组)和JTSX高剂量组(JTSX2组),另设正常组作对照。相应药物干预8周后,检测空腹血糖(FBG)、血脂、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)变化。用透射电镜观察心肌组织超微结构; TUNEL法检测心肌细胞凋亡水平;用RT-PCR和免疫组化分别检测c-JUN氨酸端激酶(c-JNK)和糖调节蛋白78(GRP78)的mRNA转录及其功能蛋白的表达水平。结果:与模型组相比,各给药组均能明显降低FBG(P <0. 05),升高SOD、GSH-Px(P <0. 05);均可降低TG、LDL-C(P <0. 05),升高HDL-C(P <0. 05);均可下调c-JNK、GRP78mRNA转录及其蛋白表达(P <0. 05),降低心肌细胞凋亡指数(AI)(P <0. 05),并改善心肌组织超微结构;与西药组相比,JTSX2疗效更显著(P <0. 05)。结论:JTSX能改善糖尿病代谢紊乱,提高抗氧化应激能力,抑制内质网应激细胞凋亡通路,从而改善心肌重构,疗效具有剂量依赖性。
Objecive:To investigate the molecular mechanism of Jiangtang Shuxin( JTSX) Recipe inhibiting endoplasmic reticulum stress c-JNK apoptotic pathway and reducing myocardial remodeling in SD diabetic rats. Methods: Type 2 diabetes were established in male SD rats by injecting streptozotocin and feeding high lipid diet. After T2 DM models established successfully,the rats were divided into model group,Western medicine group( Gliquidone + Benazepril),low-dose JTSX group( JTSX_1) and high-dose JTSXgroup(JTSX_2). Another normal group was used as control group. After 8 weeks of corresponding drug intervention,the levels of fasting blood glucose( FBG),serum superoxide dismutase( SOD),glutathione peroxidase( GSH-Px) were measured. The ultrastructure of myocardial tissue was observed with transmission electron microscope. TUNEL was used to detect cardiomyocyte apoptosis. The mRNA and their functional protein levels of c-Jun N-terminal kinases( c-JNK) and glucose-regulated protein78( GRP78) were determined by the method of RT-PCR and immunohistochemistry respectively. Results:Compared with model group,after drugs intervention,all administered groups could significantly decrease FPG( P < 0. 05),elevate SOD and GSH-Px activity( P < 0. 05); all could depress the levels of TG and LDL-C( P < 0. 05),and raise HDL-C( P < 0. 05);all could cut down the mRNA transcription and their functional protein expression of the c-JNK and GRP78( P < 0. 05); all could reduce cardiomyocyte apoptosis index( AI)( P < 0. 05) and improve the ultrastructure of myocardial tissue. Compared with Western medicine group,the JTSX_2 effect was more significant( P < 0. 05). Conclusion:JTSX can improve metabolic disorders in diabetes mellitus,ameliorate the ability of antioxidant stress,suppress endoplasmic reticulum stress apoptosis c-JNK pathway,and therefore improve myocardial remodeling,and the therapeutic effect was dose-dependent.
Objecive:To investigate the molecular mechanism of Jiangtang Shuxin( JTSX) Recipe inhibiting endoplasmic reticulum stress c-JNK apoptotic pathway and reducing myocardial remodeling in SD diabetic rats. Methods: Type 2 diabetes were established in male SD rats by injecting streptozotocin and feeding high lipid diet. After T2 DM models established successfully,the rats were divided into model group,Western medicine group( Gliquidone + Benazepril),low-dose JTSX group( JTSX_1) and high-dose JTSXgroup(JTSX_2). Another normal group was used as control group. After 8 weeks of corresponding drug intervention,the levels of fasting blood glucose( FBG),serum superoxide dismutase( SOD),glutathione peroxidase( GSH-Px) were measured. The ultrastructure of myocardial tissue was observed with transmission electron microscope. TUNEL was used to detect cardiomyocyte apoptosis. The mRNA and their functional protein levels of c-Jun N-terminal kinases( c-JNK) and glucose-regulated protein78( GRP78) were determined by the method of RT-PCR and immunohistochemistry respectively. Results:Compared with model group,after drugs intervention,all administered groups could significantly decrease FPG( P < 0. 05),elevate SOD and GSH-Px activity( P < 0. 05); all could depress the levels of TG and LDL-C( P < 0. 05),and raise HDL-C( P < 0. 05);all could cut down the mRNA transcription and their functional protein expression of the c-JNK and GRP78( P < 0. 05); all could reduce cardiomyocyte apoptosis index( AI)( P < 0. 05) and improve the ultrastructure of myocardial tissue. Compared with Western medicine group,the JTSX_2 effect was more significant( P < 0. 05). Conclusion:JTSX can improve metabolic disorders in diabetes mellitus,ameliorate the ability of antioxidant stress,suppress endoplasmic reticulum stress apoptosis c-JNK pathway,and therefore improve myocardial remodeling,and the therapeutic effect was dose-dependent.
引文
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[18]Darnel P,Miles J D B,Chengxue Q,et al. Phosphoinositide 3-Kinase(p110α)Gene Delivery Limits Diabetes-induced Cardiac NADPH Oxidase and Cardiomyopathy in a Mouse Model with Established Diastolic Dysfunction[J]. Clinical Science,2017,131(12):1345.
[19]Paras K,Mishra,Wei Ying. Diabetic Cardiomyopathy:An immunometabolic Perspective[M]. Frontiers in Endocrinology,2017:72.
[20]Wang WK,Lu QH,Wang X,et al. Ulinastatin attenuates diabetes-induced cardiac dysfunction by the inhibition of inflammation and apoptosis[J]. Exp Ther Med,2017,14(3):2497-2504.
[21]Choi K. Defective intracellular Ca2+signaling contributes to cardiomyopathy in Type 1 diabetic rats[J]. Am J Physiol Heart Circ Physiol,2002,283(4):398-408.
[22]赵灵燕,毕力夫,赵慧辉,等. 147例2型糖尿病患者中医辨证分型及临床指标相关性分析[J].北京中医药大学学报,2013,36(7):480-483.
[23]符显昭,黄振峰,黄文华,等.滋阴益气活血解毒法防治糖尿病合并ACS的回顾与展望[J].中国中西医结合急救杂志,2017,24(5):547-551.
[24]符显昭,喻嵘,成细华,等.降糖舒心方对MKR糖尿病鼠内质网应激CHOP凋亡信号及心肌重构的影响[J].中华中医药杂志,2014,29(6):1994-1997.
[2]Wu QQ,Xiao Y,Yuan Y,et al. Mechanisms contributing to cardiac remodelling[J]. Clin Sci(Lond),2017,131(18):2319-2345.
[3]周宇,李晶,鲍翠玉.内质网应激在糖尿病心肌病中的研究进展[J].中国药理学通报,2017,33(9):1200-1202.
[4]Xiao-meng W,Ying-cui W,Xiang-juan L,et al. BRD7 mediates hyperglycaemia-induced myocardial apoptosis via endoplasmic reticulum stress signalling pathway[J]. J Cell Mol Med,2017,21(6):1094-1105.
[5]Othman AI,El-Sawi MR,El-Missiry MA,et al. Epigallocatechin-3-gallate protects against diabetic cardiomyopathy through modulating the cardiometabolic risk factors,oxidative stress,inflammation,cell death and fibrosis in streptozotocinnicotinamide-induced diabetic rats[J]. Biomed Pharmacother,2017,94:362-373.
[6]符显昭,许靖,黄文华,等.糖尿病冠心病活血解毒疗法思路的构建[J].中国中医急症,2014,23(11):2024-2027.
[7]符显昭,黄月凤,王清礼,等.降糖舒心方对糖尿病合并慢性心力衰竭的影响[J].中国中西医结合急救杂志,2017,24(2):123-126.
[8]Cai L,Kang YJ. Cell death and diabetic cardiomyopathy[J].Cardiovasc Toxicol,2003,3:219-228.
[9]Shilpa R,Pradeep K S,Chunghee C,et al. Salubrinal Alleviates Pressure Overload-Induced Cardiac Hypertrophy by Inhibiting Endoplasmic Reticulum Stress Pathway[J]. Molecules and Cells,2017,40(1):66-72.
[10]Zhang C,Syed TW,Liu R,et al. Role of Endoplasmic Reticulum Stress,Autophagy,and Inflammation in Cardiovascular Disease[J]. Front Cardiovasc Med,2017,12(4):29.
[11]刘中勇,李林,方家.真武汤对心力衰竭模型大鼠心室重构及心肌细胞凋亡、纤维化的影响[J].中医杂志,2017,58(14):1218-1222.
[12]Kaneko M,Imaizumi K,Saito A,et al. ER Stress and Disease:Toward Prevention and Treatment[J]. Biol Pharm Bull,2017,40(9):1337-1343.
[13]Jing G,Xing-zhi W,Tao W,et al. Molecular signal networks and regulating mechanisms of the unfolded protein response[J]. Biomed&Biotechnol,2017,18(1):1-14.
[14]关丽英,许彩民,潘华珍.内质网应激介导的细胞凋亡[J].生物化学与生物物理进展,2007,11:1136-1141.
[15]Hong F,Liu B,Wu BX,et al. CNPY2 is a key initiator of the PERK-CHOP pathway of the unfolded protein response[J].Nat Struct Mol Biol,2017,24(10):834-839.
[16]Yang L,Zhao D,Ren J,et al. Endoplasmic reticulum stress and protein quality control in diabetic cardiomyopathy[J]. Biochim Biophys Acta,2015,1852(2):209-218.
[17]Brownlee M. Biochemistry and molecular cell biology of diabetic complications[J]. Nature,2001,414(6865):813-820.
[18]Darnel P,Miles J D B,Chengxue Q,et al. Phosphoinositide 3-Kinase(p110α)Gene Delivery Limits Diabetes-induced Cardiac NADPH Oxidase and Cardiomyopathy in a Mouse Model with Established Diastolic Dysfunction[J]. Clinical Science,2017,131(12):1345.
[19]Paras K,Mishra,Wei Ying. Diabetic Cardiomyopathy:An immunometabolic Perspective[M]. Frontiers in Endocrinology,2017:72.
[20]Wang WK,Lu QH,Wang X,et al. Ulinastatin attenuates diabetes-induced cardiac dysfunction by the inhibition of inflammation and apoptosis[J]. Exp Ther Med,2017,14(3):2497-2504.
[21]Choi K. Defective intracellular Ca2+signaling contributes to cardiomyopathy in Type 1 diabetic rats[J]. Am J Physiol Heart Circ Physiol,2002,283(4):398-408.
[22]赵灵燕,毕力夫,赵慧辉,等. 147例2型糖尿病患者中医辨证分型及临床指标相关性分析[J].北京中医药大学学报,2013,36(7):480-483.
[23]符显昭,黄振峰,黄文华,等.滋阴益气活血解毒法防治糖尿病合并ACS的回顾与展望[J].中国中西医结合急救杂志,2017,24(5):547-551.
[24]符显昭,喻嵘,成细华,等.降糖舒心方对MKR糖尿病鼠内质网应激CHOP凋亡信号及心肌重构的影响[J].中华中医药杂志,2014,29(6):1994-1997.