Zeng Xi, Huang Ping, Chen Mingkai, Liu Shiqian, Wu Nannan, Wang Fang, Zhang Jing
Department of Gastroenterology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.
Department of Gastroenterology, People's Hospital of Yichang Center, Yichang, Hubei 443003, P.R. China.
Exp Ther Med. 2018 Jan;15(1):1062-1068. doi: 10.3892/etm.2017.5466. Epub 2017 Nov 8.
The aim of the present study was to elucidate the effect of transmembrane protein 16A (TMEM16A) on portal vein smooth muscle cell (PVSMC) proliferation associated with portal vein remodeling in portal hypertension (PHT). Sprague-Dawley rats were subjected to bile duct ligation to establish a rat model of liver cirrhosis and PHT. Sham-operated animals served as controls. At 8 weeks after bile duct ligation, the extent of liver fibrosis and the portal vein wall thickness were assessed using hematoxylin-eosin staining. The protein expression levels of TMEM16A, extracellular signal-regulated kinase 1 and 2 (ERK1/2) and phosphorylated ERK1/2 (p-ERK1/2) in the portal vein were detected by immunohistochemistry and western blotting. , the lentivirus vectors were constructed and transfected into PVSMCs to upregulate the expression of TMEM16A. Isolated rat primary PVSMCs were treated with a small molecule inhibitor of TMEM16A, T16A-inhA01. Cell cycle was detected by flow cytometry. The activity of TMEM16A in the portal vein isolated from bile duct ligated rats was decreased, while the expression level of p-ERK1/2 was increased. However, , upregulation of TMEM16A promoted the proliferation PVSMCs, while inhibition of TMEM16A channels inhibited the proliferation of PVSMCs. The results indicated that TMEM16A contributes to PVSMCs proliferation , but , it may be a negative regulator of cell proliferation influenced by numerous factors.
本研究的目的是阐明跨膜蛋白16A（TMEM16A）对门静脉高压症（PHT）中与门静脉重塑相关的门静脉平滑肌细胞（PVSMC）增殖的影响。将Sprague-Dawley大鼠进行胆管结扎以建立肝硬化和PHT大鼠模型。假手术动物作为对照。胆管结扎8周后，使用苏木精-伊红染色评估肝纤维化程度和门静脉壁厚度。通过免疫组织化学和蛋白质印迹法检测门静脉中TMEM16A、细胞外信号调节激酶1和2（ERK1/2）以及磷酸化ERK1/2（p-ERK1/2）的蛋白表达水平。此外，构建慢病毒载体并转染到PVSMCs中以上调TMEM16A的表达。用TMEM16A的小分子抑制剂T16A-inhA01处理分离的大鼠原代PVSMCs。通过流式细胞术检测细胞周期。胆管结扎大鼠分离的门静脉中TMEM16A的活性降低，而p-ERK1/2的表达水平升高。然而，上调TMEM16A促进PVSMCs增殖，而抑制TMEM16A通道则抑制PVSMCs增殖。结果表明，TMEM16A促进PVSMCs增殖，但其可能是受多种因素影响的细胞增殖负调节因子。
