红景天苷对缺氧复氧星形胶质细胞细胞损伤的影响

Effects of salidroside on cell damage in anoxia reoxygenation astrocytes

目的:研究红景天苷对缺氧复氧星形胶质细胞分泌炎症因子及氧化损伤得的影响。方法:分离培养大鼠皮质星形胶质细胞,构建缺氧复氧星形胶质细胞损伤模型,在缺氧复氧前给予红景天苷预处理,CCK8测定细胞活力,二硝基苯肼法检测乳酸脱氢酶(LDH)漏出率,黄嘌呤氧化法检测超氧化物歧化酶(SOD)活性,改良二硫双硝基苯甲酸定量法检测还原型谷胱甘肽(GSH)活性,代巴比妥酸法检测丙二醛(MDA)含量,ELISA法检测细胞分泌肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6表达水平,流式细胞术检测细胞凋亡变化,Western blot检测细胞中活化型Caspase-3(Cleaved Caspase-3)和活化型Caspase-9(Cleaved Caspase-9)蛋白表达水平,用罗丹明123法检测细胞线粒体膜电位变化。结果:缺氧复氧处理星形胶质细胞后细胞活力降低,LDH漏出率升高,细胞SOD活性、GSH活性降低,MDA含量升高,细胞分泌的TNF-α、IL-1β、IL-6增多,细胞凋亡率升高,Cleaved Caspase-3和Cleaved Caspase-9蛋白水平也升高,线粒体膜电位降低。红景天苷处理后的缺氧复氧星形胶质细胞,细胞活力升高,细胞LDH漏出率降低,细胞SOD活性、GSH活性升高,MDA含量降低,细胞分泌的TNF-α、IL-1β、IL-6减少,细胞凋亡率降低,Cleaved Caspase-3和Cleaved Caspase-9蛋白表达水平降低,线粒体膜电位升高。结论:红景天苷对缺氧复氧星形胶质细胞氧化损伤、炎症因子分泌及细胞凋亡均有抑制作用,红景天苷具有减轻缺氧复氧星形胶质细胞损伤作用。

Objective: To study the effects of salidroside on inflammatory factors and oxidative damage in astrocytes exposed to hypoxia and reoxygenation. Methods: Rat cortical astrocytes were cultured to construct an anoxia reoxygenation astrocyte injury model.Salidroside treatment was given before anoxic reoxygenation.CCK8 assay detected cell viability.The leakage rate of LDH was detected by two nitrophenyl hydrazine method.Xanthine oxidation assay was used to detect SOD activity.Modified two sulfur double nitrobenzoic acid quantitative method was used to detect GSH activity.MDA content was detected by barbituric acid method.ELISA assay was used to detect the protein expression levels of TNF-α,IL-1β and IL-6.Flow cytometry was used to detect cell apoptosis.Western blot was used to detect the protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 in the cells,and the mitochondrial membrane potential was detected by Luo Danming 123 assay. Results: The activity of cell survival rate in anoxia reoxygenation astrocytes was decreased.The leakage rate of LDH was increased.The activity of GSH and SOD was reduced,while the content of MDA was increased.The expression of TNF-α,IL-1 β and IL-6 secreted by cells were increased.The rate of apoptosis increased.Cleaved Caspase-3 and Cleaved Caspase-9 protein expression was also increased,and the mitochondrial membrane potential was decreased.Anoxia and reoxygenation astrocytes were treated with salidroside.Cell viability was elevated,the leakage rate of LDH was reduced.The activity of GSH and SOD was elevated.The content of MDA was reduced.The expressions of TNF-α,IL-1β and IL-6 were induced in cells.The rate of apoptosis was reduced.The protein expression levels of Cleaved Caspase-3 and Cleaved Caspase-9 were decreased,while the mitochondrial membrane potential was increased. Conclusion: Salidroside inhibits oxidative damage,secretion of inflammatory cytokines and apoptosis of astrocytes in hypoxia reoxygenation.Salidroside can alleviate the damage of astrocytes induced by anoxia reoxygenation.