Department of Cardiovascular Surgery, Second Hospital of Shandong University, Jinan, P. R. China.
J Cell Biochem. 2018 Jan;119(1):806-816. doi: 10.1002/jcb.26243. Epub 2017 Aug 30.
The study explores the effect of astragalus polysaccharide (APS) mediating P13K/Akt/eNOS signaling pathway on apoptosis of myocardial microvascular endothelial cells (MMECs) in hypoxia/reoxygenation (H/R). MMECs were classified into blank, H/R, H/R + 25 mg/L APS, H/R + 50 mg/L APS, H/R + 100 mg/L APS, H/R + LY, and HR + 100 mg/L APS + LY groups. Cell viability was detected using MTT assay and apoptotic cell morphological changes by Hoechst staining. NO content, cell cycle and apoptosis, PI3K/Akt/eNOS signaling pathway proteins were detected using nitrate reductase assay, flow cytometry and Western blotting. An increased cell survival rate, NO content and expression of PI3K/Akt/eNOS signaling pathway associated proteins, and a decreased apoptosis rate was observed in the H/R + 50 mg/L APS and H/R + 100 mg/L APS groups compared with the H/R and H/R + 25 mg/L APS groups. Compared with the H/R + 50 mg/L APS group, the apoptosis rate decreased, whereas the cell survival rate, NO content and expression of PI3K/Akt/eNOS signaling pathway associated proteins increased in the H/R + 100 mg/L APS group. The H/R + LY and HR + 100 mg/L APS + LY groups followed opposite trends. In comparison to the HR + 100 mg/L APS group, the apoptosis rate in the H/R + LY and HR + 100 mg/L APS + LY groups increased, and the cell survival rate, NO content and expression of PI3K/Akt/eNOS signaling pathway associated proteins decreased. Collectively, APS improves the damage caused by H/P by mediating PI3K/Akt/eNOS signaling pathway.
该研究探讨了黄芪多糖(APS)通过调节 P13K/Akt/eNOS 信号通路对缺氧/复氧(H/R)心肌微血管内皮细胞(MMECs)凋亡的影响。将 MMECs 分为空白组、H/R 组、H/R+25mg/L APS 组、H/R+50mg/L APS 组、H/R+100mg/L APS 组、H/R+LY 组和 H/R+100mg/L APS+LY 组。采用 MTT 法检测细胞活力,Hoechst 染色观察细胞凋亡形态变化。硝酸还原酶法、流式细胞术和 Western blot 法检测 NO 含量、细胞周期和凋亡、PI3K/Akt/eNOS 信号通路蛋白。与 H/R 组和 H/R+25mg/L APS 组相比,H/R+50mg/L APS 组和 H/R+100mg/L APS 组细胞存活率升高,NO 含量和 PI3K/Akt/eNOS 信号通路相关蛋白表达升高,细胞凋亡率降低。与 H/R+50mg/L APS 组相比,H/R+100mg/L APS 组细胞凋亡率降低,细胞存活率、NO 含量和 PI3K/Akt/eNOS 信号通路相关蛋白表达升高。H/R+LY 组和 H/R+100mg/L APS+LY 组呈相反趋势。与 H/R+100mg/L APS 组相比,H/R+LY 组和 H/R+100mg/L APS+LY 组细胞凋亡率升高,细胞存活率、NO 含量和 PI3K/Akt/eNOS 信号通路相关蛋白表达降低。综上所述,APS 通过调节 PI3K/Akt/eNOS 信号通路改善 H/R 引起的损伤。