基础研究(Basic Research)

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

Published at: 2017年第26卷第2期

李克跃 1,2 , 石承先 2 , 汤可立 2 , 刘振华 2 , 黎涛 2 , 张帅民 2 , 徐贤刚 2
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
通讯作者 承先 石 Email: chengxianL@aliyun.com
DOI: 10.3978/j.issn.1005-6947.10.3978/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。


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

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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引用

引用本文: 克跃 李, 承先 石, 可立 汤, 振华 刘, 涛 黎, 帅民 张, 贤刚 徐. 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 [J]. 中国普通外科杂志, 2017, 26(2): 172-178.
Cite this article as: LI Keyue, SHI Chengxian, TANG Keli, LIU Zhenhua, LI Tao, ZHANG Shuaimin, XU Xiangang . 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 [J]. Chin J Gen Surg, 2017, 26(2): 172-178.