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目的:探讨TNF-α对兔胆管成纤维细胞P311/TGF-β1/α-SMA通路的影响及川芎嗪的干预作用。方法:分离、培养正常家兔胆管成纤维细胞并鉴定,将胆管成纤维细胞分别给予TNF-α、TNF-α联合不同浓度的TMP(0.08、0.4、2.0 mg/m L)干预48 h,以无处理的胆管成纤维细胞为空白对照,用CCK-8法检测细胞增殖水平;real-time PCR检测细胞P311、TGF-β1、α-SMA m RNA表达;Western blot检测细胞TGF-β1、α-SMA蛋白表达。结果:与空白对照比较,胆管成纤维细胞经TNF-α处理后,增殖明显增强,P311、TGF-β1、α-SMA m RNA以及TGF-β1、α-SMA蛋白表达明显上调(均P<0.05);TMP对TNF-α的上述效应有抑制作用,并且呈现浓度依耐趋势,其中0.4、2.0 mg/m L的TMP有明显作用(均P<0.05)。结论:TNF-α可能通过调控P311/TGF-β1/α-SMA信号通路促进胆管成纤维细胞增殖,TMP能抑TNF-α对该通路的活化,故可能对良性胆道狭窄有防治作用。
Objective: To investigate the effect of TNF-α on P311 / TGF-β1 / α-SMA pathway in rabbit bile duct fibroblasts and the effect of ligustrazine. Methods: Normal rabbit bile duct fibroblasts were isolated and cultured. The bile duct fibroblasts were treated with TNF-α and TNF-α for 48 h with different concentrations of TMP (0.08,0.4,2.0 mg / mL) The untreated bile duct fibroblasts were blank control, the cell proliferation was detected by CCK-8 method; the expression of P311, TGF-β1 and α-SMA m RNA were detected by real-time PCR; the expressions of TGF- SMA protein expression. Results: Compared with the blank control group, the proliferation of biliary fibroblasts significantly increased after treatment with TNF-α, and the expressions of P311, TGF-β1, α-SMA m RNA and TGF-β1 and α- ). TMP inhibited the above effects of TNF-α, and showed a trend of concentration-dependent. TMP of 0.4 and 2.0 mg / mL had a significant effect (all P <0.05). CONCLUSION: TNF-α may promote bile duct fibroblast proliferation through the regulation of P311 / TGF-β1 / α-SMA signaling pathway. TMP inhibits the activation of TNF-α pathway and may prevent and treat benign biliary stricture.