Relationship between rat hepatic fibrosis and regulation of hepatic stellate cell autophagy by microRNA-181a |
| |
| Authors: | CHEN Jing ZHENG Qi CHEN Wei LAI Rui-min ZHU Yue-yong |
| |
| Institution: | Liver Research Center， The First Affiliated Hospital， Institute for Liver Disease， Fujian Medical University， Fuzhou 350005， China |
| |
| Abstract: | AIM To observe the changes of liver structure， the levels of transforming growth factor-β1 （TGF-β1）， microRNA-181a， LC3-II/-I， beclin-1 and collagen deposition in hepatic fibrosis （HF） rats induced by carbon tetrachloride （CCl4）， and the effect of microRNA-181a on autophagy of rat hepatic stellate cells （HSCs） induced by TGF-β1， and to explore the possible mechanism of microRNA-181a in regulating HSC activation and HF. METHODS Wistar rats （n=40） were randomly divided into 5 groups （with 8 in each）： control group （subcutaneous injection of olive oil， 3 mL/kg， twice a week）， and CCl4-induced HF groups of 2， 4， 6 and 8 weeks （subcutaneous injection of 40% CCl4， 3 mL/kg， twice a week for 2， 4， 6 and 8 weeks， respectively）. Masson staining was used to evaluate the changes of HF in rats. The levels of TGF-β1 in serum and liver tissue of the rats were measured by ELISA. The level of microRNA-181a in rat liver tissues was detected by RT-qPCR. The protein levels of LC3-II/-I， beclin-1， α-smooth muscle actin （α-SMA）， collagen type I （Col I） and collagen type Ⅲ （Col Ⅲ） in rat liver tissues were measured by Western blot. HSC-T6 cells were transfected with microRNA-181a inhibitor， or pretreated with the autophagy inhibitor 3-methyladenine （3-MA）， before treatment with TGF-β1 to stimulate autophagy. The expression of microRNA-181a， LC3-II/-I， beclin-1， α-SMA， Col I and Col Ⅲ in HSC-T6 cells were determined by RT-qPCR and Western blot. RESULTS The levels of TGF-β1， microRNA-181a， LC3-II/-I ratio and beclin-1 in liver tissues showed an overall trend of increasing with the progression of HF， and microRNA-181a expression showed a positive correlation with autophagy-associated proteins （P<0.01）. MicroRNA-181a level was significantly increased， which was associated with TGF-β1-induced autophagy and activation of HSC-T6 cells.MicroRNA-181a expression was significantly down-regulated in the HSC-T6 cells transfected with microRNA-181a inhibitor， along with suppression of autophagy and cell activation （P<0.01）， which were similar to the effects of 3-MA treatment. CONCLUSION CCl4 promotes rat HF， the microRNA-181a expression of liver tissue， and autophagy in a time-dependent manner. Reducing the expression of microRNA-181a in HSC-T6 cells inhibits the autophagy of HSCs-T6 cells induced by TGF-β1. The regulation of HSC autophagy by microRNA-181a may be involved in rat HF. |
| |
| Keywords: | MicroRNA-181a Hepatic stellate cells Autophagy Hepatic fibrosis Transforming growth factor-β1 |
|
| 点击此处可从《园艺学报》浏览原始摘要信息 |
| 点击此处可从《园艺学报》下载免费的PDF全文 |
