瓜蒌皮激活PI3K/PKB通路减轻柯萨奇病毒感染心肌细胞损伤的实验研究  

Pericarpium Trichosanthis Reduces Damage to Myocardial Cells Infected by the Coxsackie Virus by Activating the PI3K/PKB Pathway

作　　者：邱清勇 刘大勇 彭锦 吕秋 杜兰雪 QIU Qingyong;LIU Dayong;PENG Jin;LYU Qiu;DU Lanxue(928th Hospital Of PLA Joint Locistics Support Force,Haikou 570206,China)

机构地区：[1]中国人民解放军联勤保障部队第九二八医院,海口570206

出　　处：《病毒学报》2021年第5期1036-1042,共7页Chinese Journal of Virology

摘　　要：柯萨奇病毒B3(Coxsackievirus B3,CVB3)是引起病毒性心肌炎最常见的病毒株,感染心肌细胞后能够激活细胞凋亡、引起细胞损伤;已有研究证实,瓜蒌皮激活磷脂酰肌醇3⁃激酶(Phosphatidylinositol 3⁃kinase,PI3K)/蛋白激酶B(Protein kinase B,PKB)通路减轻缺氧/复氧引起的心肌细胞凋亡,但瓜蒌皮对CVB3感染引起心肌细胞凋亡的调节作用及机制尚不清楚。因此,本研究将观察瓜蒌皮激活PI3K/PKB通路减轻柯萨奇病毒感染心肌细胞损伤的作用。在培养心肌H9c2细胞后分组,对照组给予不含药物及病毒的培养基,CVB3组给予含有CVB3病毒液的培养基,瓜蒌皮组含有CVB3病毒液及瓜蒌皮提取物的培养基,瓜蒌皮+抑制剂组含有CVB3病毒液、瓜蒌皮提取物及抑制剂LY294002的培养基。检测培养基中乳酸脱氢酶(Lactic dehydrogenase,LDH)及磷酸肌酸激酶同工酶(Creatine phosphokinase⁃isoenzyme⁃MB,CK⁃MB)的含量、细胞活力A490、细胞凋亡率、细胞中cleaved caspase⁃9、cleaved caspase⁃3、p⁃PI3K、p⁃PKB的表达。结果显示,与对照组比较,CVB3组培养基中LDH、CK⁃MB的含量、细胞凋亡率、细胞中天冬氨酸蛋白水解酶⁃9(cleaved caspase⁃9,cleaved cysteinyl aspartate specific proteinase⁃9)、cleaved caspase⁃3的表达均增加,A490及细胞中p⁃PI3K、p⁃PKB的表达均降低(P<0.05);与CVB3组比较,瓜蒌皮组培养基中LDH、CK⁃MB的含量、细胞凋亡率、细胞中cleaved caspase⁃9、cleaved caspase⁃3的表达均降低,A490及细胞中p⁃PI3K、p⁃PKB的表达均增加(P<0.05);与瓜蒌皮组比较,瓜蒌皮+抑制剂组培养基中LDH、CK⁃MB的含量、细胞凋亡率、细胞中cleaved caspase⁃9、cleaved caspase⁃3的表达均增加,A490及细胞中p⁃PI3K、p⁃PKB的表达均降低(P<0.05)。以上结果表明,瓜蒌皮提取物能够减轻CVB3感染心肌细胞的损伤及凋亡,激活PI3K/PKB通路是可能的分子机制。本研究首次发现瓜蒌皮提取物对CVB3感染�Coxsackie virus B3(CVB3)is the most common viral strain to cause viral myocarditis.It can activate apoptosis and cause cell injury after infecting cardiomyocytes.It has been demonstrated that Pericarpium Trichosanthis(PT)can reduce the apoptosis of cardiomyocytes caused by oxygen hypoxia/reoxygenation by activating the phosphatidylinositol 3⁃kinase(PI3K)/protein kinase B(PKB)pathway.However,the regulatory effect and mechanism of action of PT on cardiomyocyte apoptosis caused by CVB3 infection is not known.We investigated the effect of PT on reducing damage to myocardial cells infected by CVB3.The cardiomyocyte line H9c2 was cultured and divided into groups.The control group was given medium without drugs or viruses.The CVB3 group was given medium containing a solution of CVB3 virus.The PT group was given medium containing a solution of CVB3 virus and a PT extract.The PT+inhibitor group was given medium containing a solution of CVB3 virus,PT extract,and an inhibitor(LY294002).The contents of lactate dehydrogenase(LDH)and creatine kinase⁃myocardial band(CK⁃MB),absorbance at 490 nm(A490),percent apoptosis,as well as expression of cleaved caspase⁃9,cleaved caspase⁃3,phosphorylated(p)⁃PI3K and p⁃PKB were measured.Compared with the control group,the contents of LDH and CK⁃MB,percent apoptosis,and expression of cleaved caspase⁃9 and cleaved caspase⁃3 increased,and A490,and expression of p⁃PI3K and p⁃PKB in cells decreased in the CVB3 group(P<0.05 for all).Compared with the CVB3 group,the contents of LDH and CK⁃MB,percent apoptosis,and expression of cleaved caspase⁃9 and cleaved caspase⁃3 decreased,and A490,and expression of p⁃PI3K and p⁃PKB in cells increased in the PT group(P<0.05 for all).Compared with the PT group,the contents of LDH and CK⁃MB,percent apoptosis,and expression of cleaved caspase⁃9 and cleaved caspase⁃3 increased,and A490,and expression of p⁃PI3K and p⁃PKB in cells decreased in the PT+inhibitor group(P<0.05 for all).These results showed that a PT extract coul

关 键 词：病毒性心肌炎 柯萨奇病毒B3(CVB3) 瓜蒌皮 凋亡 PI3K/PKB通路 

分 类 号：R373.23[医药卫生—病原生物学]