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
作者: |
1,2李克跃,
2石承先,
2汤可立,
2刘振华,
2黎涛,
2张帅民,
2徐贤刚
1 天津医科大学研究生院,天津 300070 2 贵州省人民医院 肝胆外科,贵州 贵阳 550002 |
通讯: |
石承先
Email: chengxianL@aliyun.com |
DOI: | 10.3978/.10.3978/j.issn.1005-6947.2017.02.007 |
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 |
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/mL)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 mRNA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 mRNA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/mL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。
关键词:
胆管
缩窄,病理性
成纤维细胞
川芎嗪
Influence of TNF-α on activity of P311/TGF-β1/α-SMA signaling pathway in rabbit bile duct fibroblasts and the interventional effect of tetramethylpyrazine
CorrespondingAuthor:SHI Chengxian Email: chengxianL@aliyun.com
Abstract
Objective: To investigate the influence of TNF-α on activity of P311/TGF-β1/α-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. The bile duct fibroblasts were exposed to TNF-α or TNF-α 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 assay, mRNA expressions of P311, TGF-β1 and 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-β1 and α-SMA as well as the protein expressions of TGF-β1 and α-SMA were significantly up-regulated compared with blank control cells (all P<0.05); TMP inhibited the above effects of TNF-α 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 via regulating P311/TGF-β1/α-SMA signaling pathway in fibroblasts, and TMP can inhibit the activation of this pathway induced by TNF-α, so it may have preventive and therapeutic effect on benign biliary stricture.
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