川芎嗪对肝星状细胞TGF -β1及其Ⅰ-Ⅱ型受体mRNA及NF-κB表达影响.doc

川芎嗪对肝星状细胞TGF -β1及其Ⅰ\Ⅱ型受体mRNA及NF-κB表达影响【摘要】 目的：探讨中药单体川芎嗪(TMP)对肝星状细胞(HSC)核因子-κB(NF-κB) 、转化生长因子-β1(TGF-β1)及其Ⅰ、Ⅱ型受体表达的影响。方法：体外培养人肝星状细胞，免疫细胞化学法检测HSC中NF-κB表达以及核迁移情况，RT-PCR法检测HSC中TGF-β1及其Ⅰ、Ⅱ型受体mRNA表达。结果：TMP组NF-κB表达量较少并且主要位于细胞质，很少发生核迁移。RT-PCR显示，与阴性对照组相比，TMP组TGF-β1 mRNA相对表达量均有显著性差异(P＜0.01)；TMP 200 mg/L、800 mg/L组TβRⅠmRNA相对表达量降低，其差异有显著性(P＜0.05或 P＜0.01)； TMP 200 mg/L、800 mg/L组TβRⅡmRNA相对表达量均有显著性差异(P＜0.01)。结论：川芎嗪可降低HSC中NF-κB表达并抑制其核转移，降低TGF-β1及其Ⅰ、Ⅱ型受体mRNA表达，表明川芎嗪可抑制HSC活化，这可能与阻断NF-κB信号通路、阻断TGF-β/Smad通路的信号传导有关。 【关键词】 川芎嗪；肝星状细胞；肝纤维化；NF-κB；TGF-β1 Effects of Tetramethylpyrazine on Expression of TGF-β1 and its receptors mRNA，NF-κB in Human Hepatic Stellate Cells Fan Yuhui1, Zhu Lin2, Hua Haiying3 (1Henan Technician Training College of Medicine;2Department of Pharmacology, First Affiliated Hospital of Zhengzhou University; 3Zhengzhou University Academy of Medical and Pharmaceutical SciencesKaifeng 475003) 【Abstract】 Objective: To observe the effects of tetramethylpyrazine(TMP) on expression of Transforming growth factor-beta l(TGF-β1)and its receptors mRNA，nuclear factor-κB(NF-κB) in human hepatic stellate cells(HSCs), to investigate the effect of TMP against hepatic fibrosis. Methods: HSCs cultured in vitro were divided into negative control group, positive control group(cultured with Matrine 500mg/L), experimental group(cultured with different concentrations of TMP ( 50mg/L、100mg/L、200mg/L、400mg/L、800mg/L). Immunocytochemical S-P method was used to detect the expression of NF-κB p65. The effect of TMP was evaluated of the level of TGF-β1, TGFRⅠand TGFRⅡmRNA by RT-PCR. Results: The positive expression of NF-κB in negative control group was in the plasma and nuclear of HSCs. In the positive control group and experimental groups, the expression of NF-κB was decreased and little nuclear transfer was observed. Compared with the negative control group, TMP groups and the positive control group of TGF-β1 mRNA expression of both significant difference (P 1.4方法 1.4.