瓜蒌皮注射液调控JNK/p38通路对缺氧/复氧诱导的心肌细胞损伤和凋亡的影响  

Influence of Gualoupi Injection on cardiomyocyte injury and apoptosis induced by hypoxia/reoxygenation through regulating c-Jun N-terminal kinase/p38 mitogen-activated protein kinases pathway

作　　者：周瑜湉[1] 居羚[1] 史伦 华丰 殷建峰[2] 周翔 周阔 Zhou Yutian;Ju Ling;Shi Lun;Hua Feng;Yin Jianfeng;Zhou Xiang;Zhou Kuo(Department of Pharmacy,Hospital of Traditional Chinese Medicine of Changzhou City,Changzhou 213000,China;不详)

机构地区：[1]常州市中医医院药学部,常州213000 [2]常州市中医医院心血管科,常州213000 [3]南京医科大学附属江苏省人民医院心内科,南京210029

出　　处：《中国循证心血管医学杂志》2024年第10期1231-1235,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：江苏省常州市第九批科技计划(应用基础研究指导性)项目(CJ20229034)。

摘　　要：目的探究瓜蒌皮注射液通过调节c-Jun N末端激酶/p38丝裂原活化蛋白激酶(JNK/p38)通路提高缺氧/复氧损伤条件下心肌细胞活力及抑制其凋亡的作用及可能机制。方法使用H9c2心肌细胞构建缺氧/复氧(H/R)模型,随后根据处理情况将细胞分为对照组、H/R组、H/R+瓜蒌皮组和H/R+瓜蒌皮+茴香霉素组。通过细胞计数试剂盒(CCK-8)检测各组细胞活力,试剂盒检测各组细胞中乳酸脱氢酶(LDH)、半胱天冬酶3(Caspase 3)活性;采用末端脱氧核苷酸转移酶介导的dUTP末端标记法(TUNEL)检测各组细胞凋亡并采用免疫印迹(WB)检测各组细胞中凋亡相关蛋白如B细胞淋巴瘤2(BCL-2)、BCL-2相关的X蛋白(BAX)、前半胱天冬蛋白3(Pro-caspase 3)、裂解的半胱天冬酶3(Cleaved-caspase 3)及JNK/p38通路蛋白表达。结果与对照组相比,H/R组的细胞活力显著降低,LDH和Caspase 3活性显著增加,细胞凋亡率升高,促凋亡蛋白BAX、Pro-caspase 3、Cleaved-caspase 3的表达上升,而抗凋亡蛋白Bcl-2的表达下降;同时,JNK/p38通路蛋白(JNK、p-JNK、p38、p-p38)的表达水平也显著升高,差异有统计学意义(P<0.05)。与H/R组相比,不同浓度的瓜蒌皮注射液处理后细胞活力均有所提升,尤其是2%瓜蒌皮注射液组表现出最佳效果(P<0.05)。与H/R组相比,H/R+瓜蒌皮组以及H/R+瓜蒌皮+茴香霉素组的细胞活力均显著升高,且两组的LDH和Caspase 3活性降低,细胞凋亡率及促凋亡蛋白(BAX、Pro-caspase 3、Cleaved-caspase 3)的表达下降,抗凋亡蛋白Bcl-2的表达上升,JNK/p38通路蛋白(JNK、p-JNK、p38、p-p38)的表达受到抑制(P<0.05)。与H/R+瓜蒌皮组相比,H/R+瓜蒌皮+茴香霉素组细胞活力受到抑制,LDH和Caspase 3活性升高;BAX、Pro-caspase 3、Cleavedcaspase 3及JNK、p-JNK、p38、p-p38的表达显著上升,而Bcl-2表达下降(P<0.05)。结论瓜蒌皮注射液能够通过抑制JNK/p38通路的激活增加细胞活力并减少细胞凋亡。Objective To investigate the effect and possible metabolism of Gualoupi Injection in improving cardiomyocyte activity and inhibiting cardiomyocyte apoptosis in the condition of hypoxia/reoxygenation(H/R)injury through regulating pathway of c-Jun N-terminal kinase/p38 mitogen-activated protein kinases(JNK/p38).Methods The model of H/R was established by using H9c2 cardiomyocytes,and then divided into control group,H/R group,H/R+Gualoupi Injection group and H/R+Gualoupi Injection+anisomycin group.The cardiomyocyte activity was detected by using cell counting kit-8(CCK-8),and activities of lactic dehydrogenase(LDH)and Caspase 3 were detected by using kit assay in all groups.The cardiomyocyte apoptosis was detected by using TUNEL,and apoptosis-related proteins,including BCL-2,BAX,Pro-caspase 3 and Cleaved-caspase 3,were detected by using Western blotting assay(WB),and expressions of JNK/p38 pathway was detected in all groups.Results The cardiomyocyte activity decreased significantly,activities of LDH and Caspase 3 increased significantly,cardiomyocyte apoptosis rate increased,expressions of BAX,Pro-caspase 3 and Cleaved-caspase increased,Bcl-2 expression decreased,expressions of JNK/p38 pathway proteins(JNK,p-JNK,p38,p-p38)increased significantly in H/R group compared with control group(P<0.05).The cardiomyocyte activity was raised in different groups treated by Gualoupi Injection compared with H/R group,especially in group treated with 2%of Gualoupi Injection(P<0.05).The cardiomyocyte activity increased significantly,activities of LDH and Caspase 3 decreased,expressions of cardiomyocyte apoptosis rate and pro-apoptosis proteins(BAX,Pro-caspase 3,Cleaved-caspase 3)decreased,and expression of anti-apoptosis protein increased,and expressions of JNK/p38 pathway proteins(JNK,p-JNK,p38,p-p38)were inhibited in H/R+Gualoupi Injection group and H/R+Gualoupi Injection+anisomycin group compared with H/R group(P<0.05).The cardiomyocyte activity was inhibited,activities of LDH and Caspase 3 increased,and expressions of BAX,Pro-c

关 键 词：JNK/p38通路 瓜蒌皮注射液 细胞凋亡 缺氧/复氧损伤 心肌保护作用 

分 类 号：R730.23[医药卫生—肿瘤]