Ji Wen-juan, Qu Qiang, Jin Ye, Zhao Lei, He Xiao-dong
Department of General Surgery, Peking Union Medical Collage Hospital, Beijing, China.
Zhonghua Yi Xue Za Zhi. 2009 Nov 17;89(42):2997-3001.
To investigate the mechanism of ursodeoxycholic acid (UDCA) in hepatocyte apoptosis using differentiated hepatocytes derived from bone marrow mesenchymal cells.
Rat bone marrow mesenchymal cell was induced into mature hepatocytes in vitro and then treated with PBS (Cont), deoxycholic acid (DCA), DCA plus UDCA (U + D) or UDCA alone (UDCA). Cell apoptosis was detected by Hoechst staining and Caspase-3 activity measurement. The mRNA expressions of p53 and Bax were measured by semi-quantitative RT-PCR and real-time RT-PCR. The Bax protein expression was detected by immunohistochemistry.
In comparison with Cont, DCA obviously induced hepatocyte apoptosis as measured by an increased cell count and a higher Caspase-3 activity (P < 0.05). These increments could be inhibited by addition of UDCA. The expressions of p53 and Bax in hepatocytes were up-regulated by DCA. These up-expressions could also be inhibited by UDCA. The DCA-induced increased count of Bax-positive cells could be reduced by UDCA.
UDCA inhibits DCA-induced hepatocyte apoptosis by down-regulating the expression of p53/Bax signal molecule.
利用源自骨髓间充质细胞的分化肝细胞，研究熊去氧胆酸（UDCA）在肝细胞凋亡中的作用机制。
将大鼠骨髓间充质细胞体外诱导为成熟肝细胞，然后分别用磷酸盐缓冲液（对照）、脱氧胆酸（DCA）、DCA加UDCA（U+D）或单独使用UDCA处理。通过Hoechst染色和Caspase-3活性测定检测细胞凋亡。采用半定量逆转录-聚合酶链反应（RT-PCR）和实时RT-PCR检测p53和Bax的mRNA表达。通过免疫组织化学检测Bax蛋白表达。
与对照相比，DCA可明显诱导肝细胞凋亡，表现为细胞计数增加和Caspase-3活性升高（P<0.05）。添加UDCA可抑制这些增加。DCA可上调肝细胞中p53和Bax的表达。UDCA也可抑制这些上调表达。UDCA可减少DCA诱导的Bax阳性细胞计数增加。
UDCA通过下调p53/Bax信号分子的表达抑制DCA诱导的肝细胞凋亡。
