人参皂苷Rg1对脂多糖诱导的PC12细胞损伤的保护作用

Protective effect of ginsenoside Rg1 on PC12 cell injury induced by lipopolysaccharide

目的探讨人参皂苷Rg1(简称Rg1)对脂多糖(LPS)诱导的大鼠肾上腺嗜铬细胞瘤(PC12)细胞损伤的保护作用及机制。方法常规培养PC12细胞后分为六组:A组作为对照,B组细胞采用LPS 5μg/mL处理,C组细胞采用Rg1 10μmol/L+LPS 5μg/mL处理,D组细胞采用Rg1 20μmol/L+LPS 5μg/mL处理,E组细胞采用Rg1 30μmol/L+LPS 5μg/mL处理,F组细胞采用佛波酯(PMA)20 ng/mL预孵育1 h后再用Rg1 10μmol/L+LPS 5μg/mL处理。采用CCK-8法检测细胞生存率,ELISA检测细胞炎症因子IL-1β、IL-6、IL-10和TNF-α水平,二硝基苯肼法测定乳酸脱氢酶(LDH)水平,荧光法检测活性氧(ROS)水平,Western blot检测核因子抑制蛋白(IκBα)和p65蛋白表达水平。结果与A组比较,B组细胞生存率和IκBα蛋白水平降低,IL-1β、IL-6、IL-10、TNF-α、LDH、ROS和p65蛋白水平升高(P<0.05)。与B组相比,Rg1能够呈浓度依赖性地提高PC12细胞生存率和IκBα蛋白水平(P<0.05),呈浓度依赖性地降低IL-1β、IL-6、IL-10、TNF-α、LDH、ROS和p65蛋白水平(P<0.05)。与C组相比,F组细胞生存率和IκBα蛋白水平降低,p65蛋白水平升高(P<0.05)。结论 Rg1通过降低细胞内的炎症因子、LDH和ROS水平保护PC12细胞免受LPS损伤;其机制可能与抑制NF-κB通路有关。

Objective To investigate the protective effect and underlying mechanism of ginsenoside Rg1(Rg1) on lipopolysaccharide(LPS)-induced damage in rat adrenal pheochromocytoma(PC12) cells.Methods PC12 cells were cultured routinely and then were divided into 6 groups.Group A was taken as the control.The cells in group B were treated with LPS 5 μg/mL,which in group C were treated with Rg1 10 μmol/L+LPS 5 μg/mL,in group D with Rg1 20 μmol/L+LPS 5 μg/mL,in group E with Rg1 30 μmol/L+LPS 5 μg/mL,and in group F with PMA 20 ng/mL for 1 hour and then treated with Rg1 10 μmol/L +LPS 5 μg/mL.The CCK-8 method was used to detect the cell survival rate, ELISA was used to detect the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10) and tumor necrosis factor alpha(TNF-α).The level of lactate dehydrogenase(LDH) was determined by dinitrophenylhydrazine method, the reactive oxygen species(ROS) level was detected by fluorescence method, the expression levels of nuclear factor inhibitor protein(IκBα) and nuclear factor-κB(NF-κB) p65 protein were determined by Western blot.Results Compared with group A,the cell survival rate and IκBα protein expression in group B were decreased, and IL-1β,IL-6,IL-10,TNF-α,LDH,ROS and p65 protein expression were increased(P<0.05).Compared with group B,Rg1 improved PC12 cell survival rate and IκBα protein expression and reduced the expressions of IL-1β,IL-6,IL-10,TNF-α,LDH,ROS and p65 protein in a concentration-dependent manner(P<0.05).Compared with group C,the cell survival rate and IκBα protein level in group F were decreased, and p65 protein level was increased(P<0.05).Conclusion Rg1 protects LPS-induced PC12 cells damage by reducing the levels of inflammatory factors, LDH and ROS in the cells.The mechanism may be related to the inhibition of the NF-κB pathway.