摘要
目的探讨高糖培养大鼠Müller系(retinal Müller cell line,rMC-1)细胞中红景天苷(salidroside,SAL)对炎症反应和细胞凋亡的抑制作用。方法用高糖(35.5 mmol·L-1)培养rMC-1细胞,CCK8法检测不同浓度红景天苷对高糖环境下rMC-1细胞活力的影响,ELISA检测红景天苷对高糖诱导的rMC-1细胞炎性细胞因子的影响,流式细胞术检测高糖培养rMC-1细胞中红景天苷对其细胞凋亡的影响,细胞免疫荧光检测红景天苷对高糖诱导的rMC-1细胞谷氨酰胺合成酶(glutamine synthetase,GS)表达的影响,同时采用Western blot检测其对凋亡相关蛋白Cleaved caspase1、Cleaved caspase3及NF-κB信号通路相关蛋白的影响。结果高糖诱导后rMC-1细胞活力显著降低,炎性细胞因子TNF-α及IL-1β表达水平明显升高,细胞凋亡率、凋亡相关蛋白Cleaved caspase1、Cleaved caspase3蛋白表达水平和NF-κB信号通路相关蛋白的表达水平明显增高;红景天苷作用后rMC-1细胞活力显著升高,炎性细胞因子TNF-α及IL-1β表达水平明显减低,细胞凋亡率、Cleaved caspase1、Cleaved caspase3蛋白表达水平和NF-κB信号通路相关蛋白表达明显降低;此外加入吡咯烷二硫代氨基甲酸酯(NF-κB信号通路抑制剂)rMC-1细胞凋亡率及Cleaved caspase1、Cleaved caspase3蛋白表达水平也相应降低。结论红景天苷对高糖诱导的rMC-1细胞具有保护作用,其作用机制主要是抑制炎症反应及其诱导的细胞凋亡,NF-κB信号通路可能参与红景天苷对细胞的保护作用。
Objective To investigate the inhibitory effect and mechanism of salidroside(SAL)on inflammatory response and apoptosis in rat retinal Müller cell line(rMC-1)induced by high glucose.Methods The rMC-1 cells were cultured with high glucose(HG,35.5 mmol·L-1).Effect of different concentration of SAL on the viability of rMC-1 cells was measured by CCK-8 assay.Effect of SAL on the levels of the expression of inflammatory cytokines in rMC-1 cells induced by HG was determined by Elisa.Effect of SAL on the rate of apoptotic cells in rMC-1 cells induced by HG was detected by flow cytometry.Effect of SAL on the expression of glutamine synthase(GS)in rMC-1 cells induced by HG was detected by immunofluorescence.Effect of SAL on the expression of proapoptotic proteins Cleaved caspcase1,3 and NF-κB signaling pathway was detected by Western blot.Results HG could decrease the viability of rMC-1 cells,increase the levels of TNF-α,IL-1βand apoptotic rate.At the same time,HG could increase the expression of proapoptotic proteins Cleaved caspcase1,3 and NF-κB signaling pathway.SAL significantly enhanced the viability of rMC-1 cells,decreased the levels of TNF-α,IL-1βand apoptotic rate.Treatment with SAL also inhibited the expression of proapoptotic proteins Cleaved caspcase-1,3 and NF-κB signaling pathway.The addition of pyrrolidine dithiocarbamic acid(the NF-κB signaling pathway inhibitor)could also suppress apoptosis and the expression of Cleaved caspase1 and Cleaved caspase 3.Conclusion SAL has protective effect on rMC-1 cells induced by HG through inhibiting apoptosis mediated by inflammatory response.The NF-κB signaling pathway may be involved in the protective effect of SAL on rMC-1 cells.
作者
赵宁
于洪丹
刘学政
ZHAO Ning;YU Hongdan;LIU Xuezheng(College of Integrative Chinese and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;School of Basic Medicine,Jinzhou Medical University,Jinzhou 121001,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2021年第4期408-414,共7页
Journal of Shenyang Pharmaceutical University
