大黄牡丹汤含药血清对脂多糖刺激的小鼠巨噬细胞RAW 264.7中TRIF和TRAM表达的影响  

Effects of Dahuangmudan Tang Drug Serum on Expression of TRIF and TRAM in Lipopolysaccharide-Stimulated RAW 264.7 Murine Macrophages

作　　者：刘军[1] 承解静[1] 杨晓燕[1] 杨洁[1] 王丽敏[1] 孙燕妮[1] 

机构地区：[1]上海中医药大学附属普陀医院急诊内科,上海200062

出　　处：《中国临床医学》2014年第2期122-125,共4页Chinese Journal of Clinical Medicine

基　　金：上海市中西结合学会基金资助项目(编号:ZXYQ-1231)

摘　　要：目的:研究大黄牡丹汤含药血清对细菌脂多糖(lipopolysaccharide,LPS)刺激的小鼠巨噬细胞株RAW264.7中Toll白介素受体相关调节因子(Toll-interleukin 1 receptor-domain-containing adapter-inducing interferon-b,TRIF)及TRIF相关的接头分子(TRIF-related adaptor molecule,TRAM)表达的影响。方法:20只SD大鼠随机分为4组:对照组以及中药高、中、低浓度组,每组5只。各组大鼠分别相应地以0.9%氯化钠液或高、中、低浓度大黄牡丹汤(13.2、6.6、3.3 g/kg)连续灌胃7 d,末次灌胃给药2 h后行心脏采血并处死,分离血清(即含药血清)。体外培养RAW264.7细胞,随机分为对照组、模型组(LPS处理组)以及中药高、中、低浓度组。除对照组外,其他组均用5μg/mL LPS刺激RAW264.7细胞24 h,3个中药组同时给予相应浓度大黄牡丹汤含药血清进行干预,分别采用RT-PCR和Western blotting法检测各组细胞中TRIF和TRAM的mRNA和蛋白的表达水平。结果:模型组细胞TRIF和TRAM的mRNA和蛋白表达水平均显著高于对照组(P<0.01),中药中、高浓度组细胞TRIF和TRAM的mRNA表达水平均显著低于中药低浓度组及模型组(P<0.01),中药中、高浓度组TRIF和TRAM的蛋白表达水平也明显低于低浓度组(P<0.05)及模型组(P<0.01)。结论:大黄牡丹汤含药血清可抑制LPS刺激后RAW264.7细胞中TRIF和TRAM的表达。Objective：To investigate the effect of Dahuangmudan Tang drug serum on the expression of Toll-interleukin 1 receptor-domain-containing adapter-inducing interferon-b （TRIF） and TRIF-related adaptor molecule （TRAM） in [ipopolysaccha- ride-stimulated RAW 264.7 murine macrophage cell line. Methods：Twenty SD rats were randomly divided into control group, Dahuangmudan Tang low concentration group, Dahuangmudan Tang moderate concentration group, Dahuangmudan Tang high concentration group, with five rats in each group. All rats underwent gastric perfusion daily with 0.9% sodium chloride solution, Dahuangmudan Tang in low, moderate or high concentrations （13.2, 6.6 and 3.3 g/kg）, respectively. Blood was drawn from rats＇ hearts 2 h after ending perfusion on the 7th day, and the serum （that is the drug serum） was isolated for testing. RAW 264. 7 cells were cultured and equally divided into control group, model group, Dahuangmudan Tang low, moderate and high concentration group. RAW 264.7 cells in all groups except the control group were stimulated with 5 μg/mL LPS, and cells in the three Dahuangmudan Tang groups were treated with different concentrations of drug serum of Dahuangmudan Tang for 24 h. The mRNA expression of TRIF and TRAM was measured by RT-PCR, and their protein expression was analyzed by Western blotting. Results： The expression levels of mRNA and protein of both TRIF and TRAM in the model group increased significantly compared with those in the control group （P〈0.01）. The expression levels of mRNAs and proteins of both TRIF and TRAM in Dahuangmudan Tang moderate and high concentration group were significantly lower than those in Dahuangmu- dan Tang low concentration group and the model group （P〈0.01）. Conclusions： Dahuangmudan Tang drug serum inhibits the expression of TRIF and TRAM in LPS-stimulated RAW 264.7 cells.

关 键 词：大黄牡丹汤 含药血清 巨噬细胞 Toll白介素受体相关调节因子 TRIF相关的接头分子 

分 类 号：R563.9[医药卫生—呼吸系统]