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.
Zhongguo Zhong Yao Za Zhi. 2018 Dec;43(23):4692-4697. doi: 10.19540/j.cnki.cjcmm.20180917.001.
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 PI3K/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 PI3K/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 PI3K/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 PI3K/Akt-eNOS signaling pathway.
本文旨在研究丝状铁线莲总黄酮（TFCD）后处理对心肌缺血再灌注损伤（MIRI）的影响及PI3K/Akt-eNOS信号通路的作用。将40只雄性SD大鼠随机分为五组：假手术组、模型组（I/R）、TFCD后处理组（TFCD）、TFCD后处理+LY294002（PI3K/Akt信号通路抑制剂）组（TFCD+LY）和LY294002组（LY）。再灌注结束时，记录血流动力学参数，采用HE染色评估心肌组织形态学变化，观察心肌梗死面积，采集血样测定血浆乳酸脱氢酶（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）和谷胱甘肽过氧化物酶（GSH-Px）的活性。采用Western blot检测Akt、p-Akt、eNOS和p-eNOS蛋白的表达，采用RT-PCR检测eNOS和诱导型一氧化氮合酶（iNOS）mRNA的表达。结果显示，与模型组相比，TFCD后处理显著改善血流动力学功能和心肌结构，减小心肌梗死面积，提高NO、eNOS、SOD和GSH-Px的含量，降低LDH、CK和MDA的含量，增加Akt和eNOS蛋白表达的磷酸化水平，eNOS和iNOS mRNA表达显著增加（P＜0.05或P＜0.01）。PI3K/Akt信号通路阻滞剂LY294002抑制了这些作用。上述实验表明，TFCD后处理可显著减轻大鼠MIRI，其机制可能与增强抗氧化作用、清除氧自由基、调节NO生成及激活PI3K/Akt-eNOS信号通路有关。
