尾加压素Ⅱ对原代大鼠肝脏枯否细胞分泌TGF-β1的影响  

Study on the effect of urotensin Ⅱ on the expression of transforming growth factor-β1 of primary liver Kupffer cells in rats

作　　者：刘殿刚[1] 张育先[2] 陈辉[3] 曹占宇[4] 

机构地区：[1]首都医科大学宣武医院普外科,北京100053 [2]北京市回民医院消化科,北京100054 [3]首都医科大学附属北京朝阳医院京西院区消化科,北京100043 [4]首都医科大学附属北京天坛医院普外科,北京100050

出　　处：《临床和实验医学杂志》2016年第18期1761-1764,共4页Journal of Clinical and Experimental Medicine

基　　金：国家自然科学基金(81170408);中国博士后科学基金(2012M510094)

摘　　要：目的探讨尾加压素II(UII)对原代大鼠肝脏枯否细胞分泌转化生长因子β1(TGF-β1)表达的影响。方法分离培养的原代枯否细胞接种于24孔板内培养,以不同浓度UII刺激原代大鼠枯否细胞24 h,以1×10-6mol/L UII刺激原代大鼠枯否细胞不同时间,细胞上清液中TGF-β1蛋白含量用酶联免疫吸附试验方法测定;将原代枯否细胞分为对照组、UII组、UII+Palosuran组。各组培养24 h、48 h,检测各组上清液中TGF-β1蛋白含量,以实时定量PCR检测各组TGF-β1及TβRⅠ的基因表达。结果 UII以时间依赖方式促进枯否细胞分泌TGF-β1,在24 h达峰,48 h呈下降趋势(P<0.01)。UII以浓度依赖方式促进枯否细胞分泌TGF-β1(P<0.01)。UII的促TGF-β1分泌作用能被Palosuran所阻断(P<0.01);UII亦可促进TGF-β1及TβRⅠ的基因表达,Palosuran可抑制其高表达(P<0.01)。结论UII能够以时间及浓度依赖方式促进大鼠枯否细胞表达TGF-β1,而UII受体拮抗剂Palosuran可阻断其高表达,其机制可能与抑制TβRⅠ的表达有关。Objective Methods The isolated Kupffer cells were inoculated into 24 hole plates, which were treated with different concentrations of UⅡ for 24 h , and the untreated cells were set as controls. At different period of exposure to 10 ^- 6mol / L of UⅡ, and the levels of TGF - β1 protein in supernatant were detected by ELISA. The Kupffer cells were divided into 3 groups control group, UⅡ treated group （ exposed to 10 ^-6 mol / L of UⅡ） and Palosuran group （ exposed to 10 ^-5 mol/L Palosuran for 30 min and then added 10 ^-6mol / L of UⅡ） , after an incubation period of 24 hours, the levels of TGF -β1 protein were examined by ELISA, the expression levels of TGF -β and TβR Ⅰ mRNA were measured by real - time RT - PCR . Results The expression of TGF - β1 was induced by UⅡ in a dosage - and time - dependent manner, with a maximal effect at 24 h （ P 〈 0. 01） and at concentration of 10 5mol/L （ P 〈0. 01）. Palosuran could block UⅡ - induced TGF - β1 expression in mRNA and protein levels, and suppressed TβR Ⅰ mRNA expression （ P 〈 0. 01）. Conclusion This study suggests that TGF - β1 secretion of Kupffer cells can be upregulated by UⅡ in a dosage - and time - dependent manner. In addition, the UⅡ receptor antagonist Palosuran can block its high expression, and the mechanism may be associated with the inhibition of the expression of TpR I .

关 键 词：大鼠 枯否细胞 转化生长因子Β1 尾加压素Ⅱ 肝纤维化 

分 类 号：R575[医药卫生—消化系统]