摘要
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 influence of TNF-α on activity of P311/TGF-β1/a-SMA signaling pathway in rabbit bile duct fibroblasts and the interventional effect of tetramethylpyrazine （TMP）. Methods： Bile duct fibroblasts of rabbits were isolated and cultured, and then identified, Te bile duct fibroblasts were exposed to TNF-a or TNF-a plus different concentrations （0.08, 0.4 and 2.0 mg/mL） of TMP respectively for 48 h,using untreated bile duct fibroblasts as blank control. Afterwards, the cell proliferation was determined by CCK-8 assaymRNA expressions of P311, TGF-β1and α-SMA were measured by real-time PCR, and protein expressions of TGF-β1 and α-SMA were examined by Western blot analysis. Results： In bile duct fibroblasts after TNF-α treatment, the cell proliferation significantly accelerated, and the mRNA expressions of P311, TGF-β1and α-SMA as well as the protein expressions of TGF-β 1 and α-SMA were significantly up-regulated compared with blank control cells （all P〈0.05）; TMP inhibited the above effects of TNF-a with a concentration-dependent tendency, and the inhibitions exerted by TMP at 0.4 and 2.0 mg/mL were significant （all P〈0.05）. Conclusion： TNF-α can promote proliferation of bile duct fibroblasts possibly a-SMA signaling pathway in fibroblasts, and TMP can inhibit the activation of this it may have preventive and therapeutic effect on benign biliary stricture via regulating P311/TGF-β1 / pathway induced by TNF-α, so
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2017年第2期172-178,共7页
China Journal of General Surgery
基金
贵州省科技厅-贵州省人民医院联合基金资助项目(黔科合LH字[2016]7146)
关键词
胆管
缩窄
病理性
成纤维细胞
川芎嗪
Bile Ducts
Constriction, Pathologic
Fibroblasts
TETRAMETHYLPYRAZINE
