MicroRNA-30 Protects Against Carbon Tetrachloride-induced Liver Fibrosis by Attenuating Transforming Growth Factor Beta Signaling in Hepatic Stellate Cells

Abstract

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.

TGF-β-induced hepatocyte lincRNA-p21 contributes to liver fibrosis in mice

“…In our previous study, we found that miR-30 blunted TGF-β/Smad signaling in HSCs by targeting KLF11, which suppressed the transcription of inhibitory Smad7 in TGF-β/Smad pathway 31 . Here, we further revealed that the inhibition of KLF11 by miR-30 resulted in the upregulation of Smad7 in hepatocytes (Fig.…”

“…We previously found that hepatic miR-30s decreased in the fibrotic liver and HSC-specific upregulation of miR-30 prevented liver fibrosis 31 . Here, we examined miR-30s expressions in the hepatocyte after CCl 4 treatment and found that they decreased about 1~4-fold (Supplementary Figure S2A).…”

“…Animals were maintained under pathogen-limited conditions and had free access to rodent chow and water. Mouse liver fibrosis was induced according to previously described mothed 31 . Briefly, mice were intraperitoneally injected with of 20% CCl 4 solution in sterile mineral oil at a dose of 2.5 ml CCl 4 per kilogram body weight twice per week for three or four weeks.…”

MicroRNA‐30c suppresses the pro‐fibrogenic effects of cardiac fibroblasts induced by TGF‐β1 and prevents atrial fibrosis by targeting TGFβRII

“…Several other studies have indicated that miR‐30c is significantly reduced in a model of pathological left ventricular hypertrophy 7, 8. MiR‐30c regulates the production of ECM in TGF‐β‐dependent liver fibrosis 9, 56. MiR‐30c reduces renal fibrosis and improves renal function in diabetic nephropathy by targeting connective tissue growth factor (CTGF) 10.…”

Recombinant T2 RNase protein of Schistosoma japonicum inhibits expression of α-SMA in LX-2 cells

“…Recent research shows that various intracellular signal transduction pathways are involved in regulating the progression of activation of HSCs and hepatic fibrosis (He and Dai 2015;Pinzani and Marra 2001). Transforming growth factor-β (TGF-β) signaling pathway is the most common and important pathway of those in the development of hepatic fibrosis (Tu et al 2014(Tu et al , 2015. During the classical Smad-dependent pathway of TGF-β signaling, TGF-β and TGF-β receptors often trigger the phosphorylation of Smad2 and Smad3.…”