摘要
目的 基于受体相互作用蛋白激酶1(RIP1)/受体相互作用蛋白激酶3(RIP3)/混合谱系激酶结构域样假激酶(MLKL)信号通路探讨青蒿琥酯(ART)对高糖(HG)诱导大鼠心肌细胞(H9c2)坏死性凋亡的影响.方法 采用RIPK1抑制剂Necrostatin-1与Caspase抑制剂Z-VAD-FMK处理H9c2细胞,验证HG诱导的H9c2细胞程序性坏死;将H9c2细胞分为正常组(NC组)、HG组、青蒿琥酯低、中、高浓度组(L-ART组、M-ART组、H-ART组),分别检测细胞死亡率、氧化应激因子[乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)]、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)及磷酸化RIP1(p-RIP1)、RIP1、磷酸化RIP3(p-RIP3)、RIP3、磷酸化MLKL(p-MLKL)、MLKL蛋白表达水平.结果 ART可抑制HG诱导的H9c2细胞程序性死亡;Hoechst33342/PI染色结果显示,ART可抑制H9c2坏死性死亡;与HG组比较,L-ART组、M-ART组、H-ART组处理细胞死亡率、TNF-α、IL-6、IL-1β、LDH、MDA及p-RIP1、p-RIP3、p-MLKL表达均显著下降,SOD活性显著增加(P<0.05).结论 ART通过抑制RIP1/RIP3/MLKL信号通路,抑制高糖诱导的心肌细胞坏死性凋亡.
Abstract
Objective To discuss the influence of artesunate (ART) on necroptosis of rat cardiomyocytes (H9c2 cells) induced by high glucose (HG) based on signaling pathway of receptor interacting protein kinase 1/receptor interacting protein kinase 3/mixed lineage kinase domain-like protein (RIP1/RIP3/MLKL).Methods H9c2 cells were treated with RIPK1 inhibitor-necrosstatin-1 and caspase inhibitor-Z-VAD-FMK to verify the programmed necrosis of H9c2 cells induced by HG.H9c2 cells were divided into normal group (NC group),HG group and low-dose,mid-dose and high-dose ART groups (L-ART group,M-ART group and H-ART group).The cell mortality,oxidative stress factors[lacticdehydrogenase (LDH),malondialdehyde (MDA),superoxide dismutase (SOD)],tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),interleukin-1β (IL-1β),and protein expressions of phosphorylated RIP1 (p-RIP1),RIP1,phosphorylated RIP3 (p-RIP3),RIP3,phosphorylated MLKL (p-MLKL) and MLKL were detected respectively.Results ART could inhibit HG-induced programmed death of H9c2 cells.The results of Hoechst 33342/PI staining showed that ART could inhibit H9c2 necrotic death.The cell mortality,TNF-α,IL-6,IL-1β,LDH,MDA,and expressions of p-RIP1,p-RIP3 and p-MLKLdecreased significantly,and SOD activity increased significantly in L-ART group,M-ART group and H-ART groupcompared with HG group(P＜0.05).Conclusion ART can inhibit the necroptosis of cardiomyocytes induced by HG through restraining signaling pathway of RIP1/RIP3/MLKL in rats.
维普
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