肉豆蔻-8散通过JAK2/STAT3信号通路减轻缺氧/复氧心肌细胞的损伤
Roudoukou-8 San reduces the hypoxia/reoxygenation injury of cardiac myocytes through JAK2/STAT3 pathway
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摘要: 研究肉豆蔻-8散提取物对心肌细胞缺氧/复氧损伤的保护作用及其与酪氨酸蛋白激酶2(JAK2)/信号转导子和激活子3(STAT3)信号通路的关系。使用连二亚硫酸钠(Na2S2O4)建立H9c2细胞缺氧/复氧损伤模型；采用CCK-8法检测细胞活力；采用全自动生化分析仪检测细胞培养液中乳酸脱氢酶(LDH)、磷酸肌酸激酶(CK)和天冬氨酸氨基转移酶(AST)的含量；采用试剂盒法测定细胞中过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)的含量；采用Hoechst染色法检测细胞凋亡程度；采用蛋白免疫印迹法(Western blot)检测各组心肌细胞JAK2、p-JAK2、STAT3、p-STAT3、Bcl-2、Bax的蛋白表达。Na2S2O4对H9c2细胞活力的半数抑制量为2mmol/L；与模型组比较，不同浓度肉豆蔻-8散提取物均能提高细胞活力；能显著降低细胞培养液中CK、LDH、AST的水平、能显著增加细胞中CAT、SOD、GSH-Px的水平；同时可显著增加p-JAK2、p-STAT3、Bcl-2的蛋白表达、显著减少Bax的蛋白表达，而AG490阻断剂组可减弱肉豆蔻-8散提取物的作用。肉豆蔻-8散通过JAK2/STAT3信号通路发挥对Na2S2O4诱导缺氧/复氧损伤心肌细胞的保护作用。
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关键词:
- 肉豆蔻-8散 /
- 连二亚硫酸钠 /
- AG490阻断剂 /
- 心肌细胞缺氧/复氧损伤 /
- 酪氨酸蛋白激酶2(JAK2)/信号转导子和激活子3(STAT3)
Abstract: To study the protective effects of Roudoukou-8 San extract on hypoxia/reoxygenation injury of cardiac myocytes and its relationship with tyrosine protein kinase 2(JAK2)/signal transducer and activator 3(STAT3)signaling pathway, hypoxia/reoxygenation injury model of H9c2 cells was built by sodium dithionite(Na2S2O4). The vitality of the cells was tested by CCK-8; the contents of lactate dehydrogenase(LDH), creatine phosphate kinase(CK)and aspartate aminotransferase(AST)in cell culture medium were tested by fully automatic biochemical analyzer; the contents of catalase(CAT), superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)in cells were tested by kits; cell apoptosis degree was tested by hoechst staining. And the protein expressions of JAK2, p-JAK2, STAT3, p-STAT3, Bcl-2 and Bax in myocardial cells of each group were tested by Western blot. Hypoxia for 1 hour and reoxygenation for 3 hours of 2 mmol/L Na2S2O4 caused damage of about 50% H9c2 cells. Compared with the model group, the extracts of Roudoukou-8 San with different concentrations could increase the viability of cells. Besides, contents of CK, LDH and AST in cell culture medium were decreased significantly, while contents of CAT, SOD and GSH-Px were increased significantly. At the same time, the expression of p-JAK2, p-STAT3 and Bcl-2 were significantly increased and that of Bax was significantly decreased. The effects of Roudoukou-8 San extract could bereduced by AG490 blocker. Therefore, Roudoukou-8 San extract possesses protective effects on Na2S2O4 induced-hypoxia/reoxygenation injury of cardiac myocytes through JAK2/STAT3 signaling pathway. -
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[1] Lakota J. Molecular mechanism of ischemia-reperfusion injury after myocardial infarction and its possible targeted treatment[J].Int J Cardiol,2016,220:571-572. [2] Hashmi S,Al-Salam S.Acute myocardial infarction and myocardial ischemia-reperfusion injury:a comparison[J].Int J Clin Exp Pathol,2015,8(8):8786-8796. [3] Heusch G,Boengler K,Schulz R.Inhibition of mitochondrial permeability transition pore opening:the Holy Grail of cardioprotection[J].Basic Res Cardiol,2010,105(2):151-154. [4] Li T,Zhang P,Liu J,et al.Protective effects of hemoglobin-based oxygen carrier given to isolated heart during ischemia via attenuation of mitochondrial oxidative damage[J].Free Radic Biol Med,2010,48(8):1079-1089. [5] Akewang LD.Pu Ji Preparation (普济方集)[M].Inner Mongolia:Inner Mongolia Science and Technology Press,1986 [6] Inner Mongolia Autonomous Region Food and Drug Administration.Inner Mongolia Mongolian Medicine Preparation Specification(内蒙古蒙药制剂规范)[S].Inner Mongolia:Inner Mongolia People′s Publishing House,2014:152. [7] Xiao YF,Li WY,Wang YH.The protective effect of the eight-flavor capsule on the heart of the experimental animal[J].Chin Tradit Pat Med(中成药),2016,38(4):898-902. [8] Zhang YY,Xiao YF,Li WY,et al.Effects of Roudoukou-8 San against hydrogen peroxide-induced injury of cardiomyocyte[J].J China Pharm Univ(中国药科大学学报),2018,49(2):222-228. [9] Lochner A.Cardioprotective effect of melatonin against ischaemia reperfusion damage[J].Front Biosci,2013,E5(1):305-315. [10] Smith CCT,Dixon RA,Wynne AM,et al.Leptin-induced cardioprotection involves JAK/STAT signaling that May be linked to the mitochondrial permeability transition pore[J].Am J Physiol-Heart Circ Physiol,2010,299(4):H1265-H1270. [11] Kamiński KA,Kozuch M,Bonda TA,et al.Effect of interleukin 6 deficiency on the expression of Bcl-2 and Bax in the murine heart[J].Pharmacol Rep,2009,61(3):504-513. [12] Tanaka T,Yasui Y,Tanaka M,et al.Melatonin suppresses AOM/DSS-induced large bowel oncogenesis in rats[J].Chem Biol Interact,2009,177(2):128-136. [13] Patel AK,Hackam AS.Toll-like receptor 3(TLR3)protects retinal pigmented epithelium(RPE)cells from oxidative stress through a STAT3-dependent mechanism[J].Mol Immunol,2013,54(2):122-131. [14] Mantel C,Messina-Graham S,Moh A,et al.Mouse hematopoietic cell-targeted STAT3 deletion:stem/progenitor cell defects,mitochondrial dysfunction,ROS overproduction,and a rapid aging-like phenotype[J].Blood,2012,120(13):2589-2599.
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