川芎含药血清对肝星状细胞Toll样受体4及下游信号因子MyD88表达的影响

Effect of Ligusticum wallichii-containing serum on expressions of Toll-like receptor 4 and myeloid differentiation factor 88 in hepatic stellate cells

观察川芎含药血清对脂多糖(lipopolysaccharide,LPS)刺激的大鼠肝星状细胞(hepatic stellate cell,HSC)Toll样受体4(Toll-like receptor 4,TLR4)及下游信号因子髓样分化因子88(myeloid differentiation factor 88,My D88)表达的影响。清洁级SD大鼠给予川芎水煎剂、秋水仙碱、生理盐水连续灌胃7 d制备川芎含药血清。体外培养的肝星状细胞,除空白组外,其余各组均予LPS 1 mg·L-1刺激24 h,各组含药血清干预后于37℃,5%CO2培养箱中培养。24 h后收集细胞,分别采用半定量RTPCR和Western blot法测定TLR4,My D88的表达情况。用LPS刺激大鼠HSC后,TLR4与My D88的m RNA和蛋白的表达均明显增加(与空白对照组比较P<0.01)。川芎含药血清干预后,TLR4,My D88 m RNA和蛋白的表达明显减少(与模型组比较P<0.01或P<0.05),其中川芎含药血清中、高剂量组减少最为明显。川芎含药血清可通过抑制LPS诱导的HSCs中TLR4及下游信号因子My D88的表达,参与TLR4信号通路而发挥其抗纤维化作用。

To observe the effect of Ligusticum wallichii-containing serum on the expressions of Toll-like receptor 4 and myeloid differentiation factor 88 in hepatic stellate cells. Clean-grade SD rats were randomly divided into 5 groups and orally given L. wallichii decoction, colchicine and normal saline for 7 d to prepare L. wallichii-containing serums. Except for the blank group, all of the remaining groups were stimulated with LPS 1 mg·L^-1 for 24 h. After being intervened, the L. wallichii-containing serums were cultured in 5%CO2 incubator at 37 ℃ for 24 hours. The expression of TLR4 and MyD88 were detected by RT-PCR and Western blot. After HSC was stimulated with LPS, TLR4 and MyD88 mRNA and protein expressions were significantly higher than the blank control group（P〈0.01）. After being intervened with L. wallichii-containing serum, TLR4 and MyD88 mRNA and protein expressions were notably lower than the model group （P〈0.05 or P〈0.01）. In conclusion, L. wallichii-containing serum could regulate the TLR4 signaling pathway and show the anti-fibrosis effect by inhibiting the expression of TLR4 and MyD88 in LPS-induced HSCs.