五灵胶囊对LPS诱导鼠枯否细胞p38MAPK信号转导通路的影响

Effects of WuLing capsules on p38 mitogen-activated protein kinase signaling pathway in LPS induced rat Kupffer cells

目的:探讨五灵胶囊对脂多糖(LPS)诱导的大鼠枯否细胞(Kupffer cells,KC)p38MAPK信号转导通路的影响。方法:分离纯化KCs,60ng/ml LPS刺激建立LPS的肝细胞损伤模型;40只SD大鼠药物处理后,分离制备含药血清。实验分为四组:空白血清组、LPS+空白血清组、含药血清Ⅰ组(10.0g/kg)+LPS、含药血清Ⅱ组(6.25g/kg)+LPS。KCs产生促炎因子(I125放免法测定TNF-α、IL-6、IL-8,比色法测定NO生成量),采用Western blot法检测ERK、p-ERK、p38、p-p38、TNF-α和STAT3的蛋白水平。结果:1、空白血清+LPS组,TNF-α、IL-6、IL-8和NO浓度明显高于空白血清组;2、同空白血清+LPS组比较,含药血清Ⅰ、Ⅱ组TNF-α、IL-6、IL-8和NO水平明显降低;3、与空白血清组比较,空白血清+LPS组能上调KCs对p-ERK、P38、p-P38、STAT3和TNF-α表达(p<0.01,p<0.05),对ERK表达无影响(p>0.05)4、同空白血清+LPS组比较,含药血清Ⅰ+LPS、含药血清Ⅱ+LPS组p-p38、STAT3、TNF-α表达显著下调。结论:五灵胶囊对LPS诱导的大鼠KCs p38MAPK信号转导通路及KCs分泌促炎细胞因子TNF-a、IL-6、IL-8和NO具有抑制作用,其作用机制为防治慢性肝炎提供新的理论依据,具有潜在的临床应用前景。

Objective：To investigate the effects of WuLing capsules on p38 mitogen-activated protein kinase （p38MAPK） signaling pathway of rat Kupffer cells （KCs） induced by lipopolysaccharide （LPS）. Methods：Kupffer＇s cells were isolated and purified from rat livers and exposed to 60ng/ml LPS. The cytokines secreted from Kupffer＇ s cells were determined in the presence of WuLing Capsules. Forty SD rats were treated and drugs blood srrum was made. They were seperated into four groups： Control group, LPS group, Ⅰ group （10.0g/kgWuLing capsule）＋LPS and Ⅱ group （6.25g/kg WuLing capsule） ＋LPS The protective effects of WuLing Capsule induced by LPS stimulation of kupffer＇s cells were detected. Results：WuLing Capsules significantly reduced LPS inhibited TNF-α , IL6 and IL8 secretion of Kupffer＇s cells; And significantly reduced the expression of p-p38,STAT3,TNF-α. Conclusion：WuLing Capsule may restrain KC p38MAPK signal transduction and the secretion of proinflammatory cytokines such as TNF-α, IL-6, IL-8 and NO. The mechanism may have effects on preventing and curing chronic hepatitis, and have potential clinical prosective.