PI3K/Akt-eNOS信号通路在甘木通总黄酮后处理减轻大鼠心肌缺血再灌注损伤作用  

Effects of PI3K/Akt-eNOS signaling pathway on total flavones in Clematis filamentosa post-conditioning alleviated of myocardial ischemia-reperfusion injury in rats

作　　者：聂阳[1,2] 丁立 黄海潮[1] 许良葵[1] 赵晋[3] 杨燕军[1,2] NIE Yang;DING Li;HUANG Hai-chao;XU Liang-kui;ZHAO Jin;YANG Yan-jun(Guangdong Food and Drug Vocational College,Guangzhou 510520,China;Traditional Chinese Medical Institute of Guangdong Province,Guangzhou 510520,China;Guangdong Medical College,Dongguan 523808,China)

机构地区：[1]广东食品药品职业学院,广东广州510520 [2]广东省中药研究所,广东广州510520 [3]广东医学院,广东东莞523808

出　　处：《中国中药杂志》2018年第23期4692-4697,共6页China Journal of Chinese Materia Medica

基　　金：广东省建设中医药强省课题(20161035);广东省医学科研基金项目(A2016423);广东食品药品职业学院自然科学基金项目(2017ZR001)

摘　　要：研究甘木通总黄酮(TFCD)后处理对大鼠心肌缺血再灌注损伤(MIRI)的保护作用,及与PI3K/Akt-eNOS信号通路的关系。40只SD大鼠随机分成5组:假手术组(Sham)、缺血再灌注组(I/R)、TFCD后处理组(TFCD)、TFCD后处理联合PI3K抑制剂LY294002组(TFCD+LY)、PI3K抑制剂LY294002组(LY),每组8只。记录各组大鼠心脏血流动力学指标,HE染色观察心肌组织形态学变化,测算心肌梗死面积和大鼠血清中乳酸脱氢酶(LDH)、肌酸激酶(CK)、一氧化氮(NO)、内皮型一氧化氮合酶(eNOS)、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)含量,Western blot法检测心肌Akt,p-Akt,eNOS,p-eNOS的蛋白表达,RT-PCR法检测心肌eNOS,iNOS mRNA的表达。结果显示,与I/R组比较,TFCD组大鼠心脏血流动力学指标、心肌组织形态结构明显改善,心肌梗死面积显著降低,血清中LDH,CK和MDA含量显著降低,NO,eNOS,SOD和GSH-Px含量显著升高,Akt和eNOS磷酸化水平明显增强,eNOS和iNOS mRNA表达显著提高,LY294002可显著取消TFCD的以上作用(P<0.05或P<0.01)。提示TFCD后处理能够减轻大鼠MIRI,其作用机制可能与抗氧化、清除氧自由基、调控NO生成和激活PI3K/Akt-eNOS信号通路有关。The aim of this paper was to study the effect of total flavones of Clematis filamentosa Dunn(TFCD) post-conditioning against myocardial ischemia-reperfusion injury(MIRI) and the role of PI3 K/Akt-eNOS signaling pathway. Forty male SD rats were divided randomly into five groups: Sham group, model group(I/R), TFCD post-conditioning group(TFCD), TFCD post-condition-ing+LY294002(a PI3 K/Akt signaling pathway inhibitor) group(TFCD+LY), and LY294002 group(LY). At the end of reperfusion,hemodynamic parameters were recorded,morphology changes of myocardial tissue were evaluated by using HE staining, and myocardial infarct size were observed, blood samples were obtained to determine plasma activation of lactate dehydrogenase(LDH), creatine kinase(CK) nitric oxide(NO), endothelial nitric oxide synthase(eNOS), superoxide dismutase(SOD), maleic dialdehyde(MDA) and glutathione peroxidase(GSH-Px). The expressions of Akt, p-Akt, eNOS and p-eNOS proteins were assessed by using Western blot, and eNOS and inducible nitric oxide synthase(iNOS) mRNA was measured by RT-PCR. The results showed that, compared with the model group, TFCD post-conditioning remarkably improved hemodynamics function and myocardial structure,reduced myocardial infarct size and enhanced the contents of NO, eNOS, SOD and GSH-Px, and decreased the contents of LDH, CK and MDA, increased the levels of phosphorylation of Akt and eNOS protein expression, eNOS and iNOS mRNA expression significantly(P<0.05 or P<0.01). These effects were inhibited by LY294002, a blocker of PI3 K/Akt signaling pathway. The above experiments indicated that TFCD post-conditioning could significantly reduce MIRI in rats, the mechanism of which may be associated with increasing antioxidation, scavenging oxygen free radicals, regulating NO generation and activating PI3 K/Akt-eNOS signaling pathway.

关 键 词：甘木通 心肌缺血再灌注损伤 后处理 PI3K/Akt-eNOS信号通路 

分 类 号：R285.5[医药卫生—中药学]