中国药理学通报2018，Vol.34Issue(9)：1218-1225,8.DOI:10.3969/j.issn.1001-1978.2018.09.008
PP242诱导多囊肾大鼠胆管上皮细胞自噬和凋亡的机制研究
Mechanism of PP242 inducing autophagy and apoptosis in cholangiocytes of polycystic kidney rats
闫文帝 1李珍玲 1刘特思 1李玉姬 1原田宪一 2任香善1
作者信息
- 1. 延边大学肿瘤研究中心,吉林 延边 133002
- 2. 金泽大学医学部病理学教研室,日本 金泽 9208640
- 折叠
摘要
Abstract
Aim To investigate the molecular mecha-nism of mTORC1/2 inhibitor PP242, which inhibiting cholangiocyte cell preliferation and cystic diliatation via inducing apoptosis and autophagy in the polycystic kid-ney ( PCK ) rats. Methods The expression of p-mTOR and p-Akt in the bile duct epithelial cells was examined by immunohistochemistry. The inhibiting effect of rapamycin and PP242 on cell proliferation ac-tivity on bile duct epithelial cells, the effect of gene si-lence on LC3, Beclin-1 and the effect of the authoph-agy-specific inhibitor 3-methyladenine (3-MA) on cell proliferation were respectively analyzed by WST-1 as-say. The expression of PI3K/Akt signaling pathway re-lated proteins, autophagy-related proteins LC3, Bec-lin-1 and clevead caspase-3, which were treated by PP242 were determined by Western blot. The effect of PP242 on apoptosis was detected by Annexin V/PI double staining and ELISA. The expression of LC3 in cytoplasm was detected by immunofluorescence. The a-bility of rat bile duct epithelial cells spheroid formation was detected by 3D cell culture method, and the cells were treated by single applied with rapamycin and ap- plied rapamycin combined with Rictor gene silencing respectively. Results The protein levels of p-Akt and p-mTOR markedly increased in the bile duct epitheli-um of PCK rats. PP242 inhibited the proliferation of bile duct epithelial cells more effectively than rapamy-cin and showed a dose-and time-dependent manner ( P<0.05 ) . PP242 significantly reduced the levels of PI3K/Akt signaling pathway-related proteins in PCK rat cholangiocytes. PP242 induced apoptosis and auto-phagy, up-regulated the levels of cleaved caspase-3, Beclin-1 and increased the ratio of LC3-II/LC3-I. The combination of Rictor gene silencing and rapamycin was more effective than rapamycin alone in inhibiting cholangiocytes in PCK rats. The inhibitory effect of PP242 on the cell viability was significantly weakened by treatment with 3-MA and knockdown of LC3 and Beclin-1 ( P <0.05 ) . Conclusions PP242 inhibits the proliferation of PCK rat cholangiocytes through PI3K/Akt/mTOR signaling pathway, and the mecha-nism is closely related with autophagy and apoptosis.关键词
自噬/凋亡/PI3K/Akt/mTOR 信号通路/多囊肾大鼠/PP242/Caroli病Key words
autophagy/apoptosis/PI3K/Akt/mTOR axis/polycystic kidey rats/PP242/Caroli’s disease分类
医药卫生引用本文复制引用
闫文帝,李珍玲,刘特思,李玉姬,原田宪一,任香善..PP242诱导多囊肾大鼠胆管上皮细胞自噬和凋亡的机制研究[J].中国药理学通报,2018,34(9):1218-1225,8.
