血根碱通过STAT3通路对LPS致RAW264.7细胞炎症的保护作用  

The Protective Effect of Sanguinarine on LPS-induced RAW264.7 Cells through the STAT3 Pathway

作　　者：张梦雅[1] 王春丽[1] 杜先华[1] 黄松[1,2] 

机构地区：[1]广州中医药大学,广东广州510006 [2]东莞广州中医药大学中医药数理工程研究所,广东东莞523000

出　　处：《中药新药与临床药理》2017年第6期714-718,共5页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金："十二五"国家科技支撑计划项目(2012BAI29B09);广东省科技计划项目(2013A022100002)

摘　　要：目的研究血根碱对脂多糖(LPS)诱导的RAW264.7细胞炎症的保护作用及其机制。方法采用MTT法检测血根碱对细胞增殖作用的影响;ELISA法检测细胞上清中肿瘤坏死因子(TNF-α)和白细胞介素-6(IL-6)的含量;RT-PCR法检测细胞中STAT3 m RNA的表达;Western Blot法检测STAT3通路JAK2、STAT3磷酸化蛋白的表达。结果血根碱在0.01μg·m L^(-1)~5μg·m L^(-1)浓度范围对RAW264.7细胞或LPS诱导的RAW264.7细胞存活率无明显影响,与正常组比较差异无统计学意义(P>0.05);与正常组比较,模型组的TNF-α和IL-6含量、STAT3 mRNA相对表达量、p-JAK2和p-STAT3蛋白相对表达量均明显上升(P<0.01);与模型组比较,血根碱1,0.1,0.01μg·m L^(-1)浓度组的TNF-α和IL-6含量、STAT3 mRNA相对表达量均明显下降(P<0.01),血根碱1,0.1μg·m L^(-1)浓度组的p-JAK2和p-STAT3蛋白相对表达量显著降低(P<0.01)。结论血根碱抑制LPS诱导的RAW264.7细胞炎症可能与通过STAT3通路调节有关。Objective To study the protective effect of sanguinarine on LPS-stimulated RAW264.7 cell line and its possible mechanism. Method The MTT assay was used to dectect the influence of the different concentration of sanguinarine on the prolifiration of the cells; enzyme linked immunosorbent assay（ELISA） was used to detect the contents of IL-6 and TNF-α in the supernatant; RT-PCR was used to measure the expression of STAT3 m RNA;Western Blot method was used to detect the expression level of protein involved in STAT3 signaling pathway. Result Sanguinarine showed no significant effect on survival rate of RAW264.7 cells or LPS-stimulated RAW264.7 cell compared with the normal group in the dose range of 0.01-5 μg·m L-1（P〈0.05）. Compared with the model group,sanguinarine（1,0.1 μg·m L-1）reduced the concernation of IL-6 and TNF-α,the m RNA level of STAT3（P〈0.01）,and decreased the expression of p-JAK2 and p-STAT3 proteins（P〈0.01）. Conclusion Sanguinarine has a positive effect on LPS-stimulated RAW264.7 cells through STAT3 pathway.

关 键 词：血根碱 RAW264.7细胞 脂多糖 肿瘤坏死因子 白细胞介素-6 STAT3信号通路 

分 类 号：R285.5[医药卫生—中药学]