摘要
目的 探讨松萝酸(UA)对缺氧/复氧(H/R)诱导的心肌细胞损伤的影响及其作用机制.方法 构建H9c2细胞H/R模型,使用0.5～64 μmol/L UA分别预处理H9c2细胞,MTT法筛选UA浓度;H9c2细胞随机分为对照组、模型组、UA低剂量组(1 μmol/L)、UA中剂量组(4 μmol/L)、UA高剂量组(16 μmol/L)和UA高剂量+Benzyl butyl phthalate(BBP)组(16 μmol/L UA+100 nmol/L BBP),检测各组细胞增殖活力、细胞凋亡情况,白介素(IL)-1β、IL-6、丙二醛(MDA)、超氧化物歧化酶(SOD)、腺嘌呤核苷三磷酸(ATP)、鞘氨醇激酶1(SphK1)、鞘氨醇-1-磷酸受体1(S1PR1)、细胞外调节蛋白激酶1/2(ERK1/2)蛋白表达水平.结果 1 μmol/L以上浓度UA可以显著提高H/R H9c2细胞增殖活力.与对照组相比,模型组细胞增殖能力和ATP、SOD含量下降,细胞凋亡率和IL-1β、IL-6、MDA、SphK1、S1PR1、ERK1/2表达增加(P＜0.05);与模型组相比,UA低、中、高剂量组细胞增殖能力和ATP、SOD含量上升,细胞凋亡率和IL-1β、IL-6、MDA、SphK1、S1PR1、ERK1/2表达下降,干预效果呈浓度依赖性(P＜0.05);BBP完全沉默UA对H/R诱导的心肌细胞的治疗作用.结论 UA减轻H/R H9c2细胞损伤与其抑制SphK1/S1P信号通路有关.
Abstract
Objective To investigate the impacts of usnic acid(UA)on myocardial cell injury induced by hypoxia/reoxygenation(H/R)and its mechanism.Methods The H/R model of H9c2 cells was constructed,H9c2 cells were pre-treated with 0.5～64 μmol/L UA,and the concentration of UA was screened by MTT method.H9c2 cells were randomly di-vided into control group,model group,low dose of UA group(1 μmol/L),medium dose of UA group(4 μmol/L),high dose of UA group(16 μmol/L),and high dose of UA+Benzyl butyl phosphate(BBP)group(16 μmol/L UA+100 nmol/L BBP).The proliferation activity,apoptosis,the level of interleukin(IL)-1β,IL-6,malondialdehyde(MDA),superoxide dismutase(SOD),adenine nucleoside triphosphate(ATP),sphingosine kinase 1(SphK1),sphingosine 1-phosphate re-ceptor 1(S1PR1),and extracellular regulated protein kinase 1/2(ERK1/2)proteins were detected in each group.Re-sults UA at concentrations above 1 μmol/L can significantly increase the proliferative activity of H/R induced H9c2 cells(P＜0.05).Compared with the control group,the cell proliferation ability and the content of ATP and SOD in the model group were decreased,the cells apoptosis rate and the expressions of IL-1β,IL-6,MDA,SphK1,S1PR1,and ERK1/2 were increased(P＜0.05).Compared with the model group,the cell proliferations ability and the content of ATP and SOD in the low,medium,and high dose UA groups were increased,the cells apoptosis rate and the expression of IL-1β,IL-6,MDA,SphK1,S1PR1,and ERK1/2 were decreased,the intervention effect was concentration dependent(P＜0.05).BBP completely silenced the therapeutic effect of UA on H/R induced myocardial cells.Conclusion UA attenu-ates the injury of H/R H9c2 cells,which is related to inhibition of the SphK1/S1P signaling pathway.
维普
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