瑞舒伐他汀抑制miR-122-5p减轻LPS诱导的神经细胞损伤

Effects of rosuvastatin on neuronal damage induced by lipopolysaccharide and possible action mechanism

目的探讨瑞舒伐他汀(rosuvastatin)对脂多糖(LPS)诱导的神经细胞损伤的作用和可能机制。方法体外培养大鼠肾上腺嗜铬细胞瘤PC-12,采用LPS诱导PC-12细胞建立脊髓损伤模型。不同浓度(0.01、0.1、1.0μmol/L)的瑞舒伐他汀作用于LPS诱导的PC-12细胞24 h或1.0μmol/L的瑞舒伐他汀作用于LPS诱导的转染miR-122-5p模拟物的PC-12细胞24 h后,CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中Cleaved-caspase-3、IL-1β、IL-6和TNF-α蛋白表达,RT-qPCR检测miR-122-5p表达。结果PC-12细胞经LPS诱导后,细胞A值降低(P<0.05),凋亡率及Cleaved-caspase-3、IL-1β、IL-6、TNF-α的蛋白表达水平升高(P<0.05),miR-122-5p表达水平升高(P<0.05)。而不同浓度(0.01、0.1、1.0μmol/L)的瑞舒伐他汀提高LPS诱导的PC-12细胞A值(P<0.05),降低细胞凋亡率及Cleaved-caspase-3、IL-1β、IL-6、TNF-α的蛋白表达及miR-122-5p表达(P<0.05)。上调miR-122-5p逆转1.0μmol/L的瑞舒伐他汀对LPS诱导的PC-12细胞增殖、凋亡及炎性因子IL-1β、IL-6、TNF-α表达的影响(P<0.05)。结论瑞舒伐他汀可能通过下调miR-122-5p抑制LPS诱导的PC-12细胞凋亡及炎性因子表达。

Objective To investigate the effects of rosuvastatin on neuronal damage induced by lipopolysaccharide(LPS),and to explore possible action mechanism.Methods The rat adrenal pheochromocytoma PC-12 cells were cultured in vitro,which were induced by LPS to establish a spinal cord injury model.After the cells were treated by different concentrations(0.01,0.1,1.0μmol/L)of rosuvastatin for 24h,or the cells transfected by miR-122-5p were treated by 1.0μmol/L of rosuvastatin for 24h,the CCK-8 was used to detect the cell proliferation,and flow cytometry was used to detect cell apoptosis,Western Blot was used to detect the expression levels of Cleaved-caspase-3,IL-1β,IL-6 and TNF-αin the cells,and RT-qPCR was used to detect the expression levels of miR-122-5p.Results After PC-12 cells were induced by LPS,the A value of the cell was significantly decreased(P<0.05),however,the apoptosis rate and expression levels of Cleaved-caspase-3 protein,IL-1β,IL-6,and TNF-αas well as miR-122-5p were significantly increased(P<0.05).The different concentrations(0.01,0.1,1.0μmol/L)of rosuvastatin increased the A value of PC-12 cells induced by LPS(P<0.05),however,which decreased cell apoptosis rate and the expression levels of Cleaved-caspase-3 protein,IL-1β,IL-6 and TNF-αas well as miR-122-5p(P<0.05).The up-regulation of miR-122-5p reversed the effects of 1.0μmol/L rosuvastatin on the proliferation,apoptosis and the expressions of inflammatory factors IL-1β,IL-6,and TNF-αin PC-12 cells induced by LPS(P<0.05).Conclusion Rosuvastatin may inhibit the cell apoptosis and inflammatory factor expression in PC-12 cells induced by LPS by down-regulating the expression of miR-122-5p.