黄芪-丹参对LPS诱导的小鼠巨噬细胞焦亡的影响

Effect of Astragalus and Salvia Miltiorrhiza on LPS Induced Apoptosis of Macrophages in Mice

目的:研究药对黄芪-丹参对LPS诱导的小鼠巨噬细胞炎症的影响,并探讨与焦亡相关的可能性的机制。方法:CCK-8法细胞增殖检测筛选诱导RAW264.7巨噬细胞活力值的最佳LPS浓度。用最佳的LPS浓度复制炎症模型,根据文献得到最佳的含药血清浓度干预造模后的巨噬细胞,24h后收集细胞上清做ELISA检测RAW264.7巨噬细胞炎症因子白细胞介素-1β(IL-1β),提取细胞蛋白用蛋白免疫印迹法(Western blot)测定TNF-!、NF-κB的水平。结果:CCK-8结果显示10μg/mL LPS作用时,细胞活力值显著最强,20%为最佳含药血清浓度。ELISA结果显示:模型组IL-1β浓度显著高于空白组(P<0.01),与模型组比较,黄芪-丹参药对组可降低IL-1β的水平(P<0.05,P<0.01)。Western blot结果显示:模型组TNF-!、NF-κB蛋白表达量均高于空白组(P<0.05,P<0.01),与模型组相比较,黄芪-丹参药对组可降低TNF-!、NF-κB蛋白表达量(P<0.05)。结论:药对黄芪-丹参抗LPS诱导巨噬细胞活化可能是通过抑制TNF-!/NF-κB信号通路的焦亡相关指标的表达水平。

Objective:To study the effect of Astragalus and Salvia miltiorrhiza on LPS induced macrophage inflammation in mice,and to explore the possible mechanism related to coke death.Methods:The best LPS concentration of RAW264.7macrophage was selected by CCK-8method.The best LPS concentration was used to replicate the inflammatory model,and the best serum concentration was obtained according to the literature.After 24hours,the supernatant of macrophages was collected for ELISA to detect the inflammatory factor IL-1β(IL-1β)of RAW264.7macrophages.The levels of TNF-!and NF-κB were measured by Western blot.Results:The results of CCK-8showed that the cell viability was the strongest when LPS was 10μg/mL,and 20%was the best serum concentration.The results of ELISA showed that the concentration of IL-1βin the model group was significantly higher than that in the blank group(P<0.01).The results of Western blot showed that the expression of TNF-!and NF-κB protein in the model group was higher than that in the blank group(P<0.05,P<0.01).Compared with the model group,the expression of TNF-!and NF-κB protein in the Astragalus Danshen group was decreased(P<0.05).Conclusion:The anti LPS effect of Astragalus and Salvia miltiorrhiza on macrophage activation may be through the inhibition of TNF-!/NF-κB signal pathway related to the expression level of charring.