MicroRNA-30 Protects Against Carbon Tetrachloride-induced Liver Fibrosis by Attenuating Transforming Growth Factor Beta Signaling in Hepatic Stellate Cells.
Transforming growth factor beta (TGF-β) is crucial for transdifferentiation of hepatic stellate cells (HSCs) and the blunting of TGF-β signaling in HSCs can effectively prevent liver fibrosis. Krüppel-like factor 11 (KLF11) is an early response transcription factor that potentiates TGF-β/Smad signaling by suppressing the transcription of inhibitory Smad7. Using a mouse model of carbon tetrachloride (CCl4)-induced liver fibrosis, we observed significant upregulation of KLF11 in the activated HSCs during liver fibrogenesis. Meanwhile, the downregulation of miR-30 was observed in the HSCs isolated from fibrotic liver. Adenovirus-mediated ectopic expression of miR-30 was under the control of smooth muscle α-actin promoter, showing that the increase in miR-30 in HSC greatly reduced CCl4-induced liver fibrosis. Subsequent investigations showed that miR-30 suppressed KLF11 expression in HSC and led to a significant upregulation of Smad7 in vivo. Mechanistic studies further confirmed that KLF11 was the direct target of miR-30, and revealed that miR-30 blunted the profibrogenic TGF-β signaling in HSC by suppressing KLF11 expression and thus enhanced the negative feedback loop of TGF-β signaling imposed by Smad7. Finally, we demonstrated that miR-30 facilitated the reversal of activated HSC to a quiescent state as indicated by the inhibition of proliferation and migration, the loss of activation markers, and the gain of quiescent HSC markers. In conclusion, our results define miR-30 as a crucial suppressor of TGF-β signaling in HSCs activation and provide useful insights into the mechanisms underlying liver fibrosis.
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Xiaolong
X
Tu
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Xiuxiu
X
Zheng
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Huanan
H
Li
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Zhipeng
Z
Cao
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Hanwen
H
Chang
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Shaoyuan
S
Luan
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Jie
J
Zhu
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China.
Jiangning
J
Chen
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China *State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China jfzhang@nju.edu.cn.
Yuhui
Y
Zang
*State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China *State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University and Jiangsu Engineering Research Center for microRNA Biology and Biotechnology, Nanjing University, Nanjing 210093, People's Republic of China jfzhang@nju.edu.cn.
Junfeng
J
Zhang
D000818:Animals; D002252:Carbon Tetrachloride Poisoning / Q000517:prevention & control*; D056486:Chemical and Drug Induced Liver Injury / Q000517:prevention & control*; D055166:Hepatic Stellate Cells / Q000473:pathology*; D008103:Liver Cirrhosis / Q000139:chemically induced* / Q000517:prevention & control*; D051379:Mice; D035683:MicroRNAs / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D035683:MicroRNAs; C534715:Mirn30d microRNA, mouse; D016212:Transforming Growth Factor beta
KLF11; Smad7; TGF-β signaling; hepatic stellate cell; liver fibrosis; miR-30