[关键词]
[摘要]
目的 研究氧化苦参碱(OM)对接受转化生长因子-β1(TGF-β1)刺激的人胰腺导管癌PANC-1细胞中Smad3/Gli1通路相关因子表达的影响。方法 以TGF-β1刺激PANC-1细胞模拟胰腺纤维化模型,并观察给予OM干预对Smad3/Gli1通路相关因子表达的影响。通过细胞转染技术将Gli1及Smad3的RNA干扰质粒转染入PANC-1细胞;通过Western blotting技术检测Smad3、Gli1和α-SMA的蛋白表达,ELISA技术检测纤连蛋白(FN)和I型胶原(CoL-I)在细胞培养上清中的表达。结果 与对照组相比,PANC-1细胞接受TGF-β1刺激后Smad3、Gli1和α-SMA的蛋白表达显著升高;与TGF-β1组相比,OM和TGF-β1共同处理组Smad3、Gli1和α-SMA的蛋白表达显著降低。转染Smad3干扰质粒后,Gli1、α-SMA、FN和CoL-I表达显著下降。与TGF-β1组比较,转染Gli1干扰质粒后加TGF-β1组α-SMA、FN和CoL-I表达显著降低。结论 OM可能通过调节PANC-1细胞TGF-β1/Smad3/Gli1通路发挥抗胰腺纤维化作用。
[Key word]
[Abstract]
Objective To investigate the effect of oxymatrine (OM) on expression of related molecules in Smad3, Gli1 signaling pathway in PANC-1 cells induced by TGF-β1. Methods TGF-β1-induced PANC-1 cells were used to establish the pancreatic fibrosis model in vitro, and observe the effects of OM pretreatment on the related molecular expression of Smad3/Gli1. Gli1 and Smad3 RNA interference plasmids were transfected into PANC-1 cells. The protein expression levels of Smad3, Gli1 and α-SMA were measured by Western blotting. The levels of fibronectin (FN) and type I collagen (CoL-I) in the supernatant of cell culture were detected by ELISA. Results Compared with the control group, the protein expressions of Smad3, Gli1 and α-SMA increased significantly in PANC-1 cells after treated with TGF-β1. The expressions of Gli1, α-SMA, FN, and CoL-I in PANC-1 cells decreased significantly after Gli1 RNA interference plasmid transfection compared with TGF-β1 induced group. The expression of Smad3, Gli1, α-SMA, FN, and CoL-I also decreased significantly in PANC-1 cells after Smad3 RNA interference plasmid transfection compared with TGF-β1 group. Conclusion OM could prevent pancreatic fibrosis by regulating TGF-β1/Smad3/Gli1 signaling pathway in PANC-1 cells.
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[基金项目]
国家自然科学基金资助项目(81500489);天津市自然科学基金联合资助青年项目(15JCQNJC45600);武警后勤学院附属医院种子基金重点项目(FYZ201508)