Smad2 increases the apoptosis of activated human hepatic stellate cells induced by TRAIL.

The activation of hepatic stellate cells (HSCs) plays a critical role in the development of liver fibrosis. The induction of apoptosis in activated HSCs during the recovery phase of hepatic fibrosis represents a potential anti-fibrotic therapy. We have previously shown that Smad2 protects against hepatic fibrogenesis; however, the role of Smad2 in the regulation of activated HSC apoptosis remains unknown. We hypothesized that Smad2 regulates the apoptosis of activated HSCs, leading to the resolution of liver fibrosis. To test this hypothesis, the livers of rats were harvested at 0 and 4 weeks after hepatic fibrosis was established by CCl4 injection. Furthermore, TGF-β1-activated HSCs were treated with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) following the silencing or overexpression of Smad2. Both the phosphorylation of Smad2 and TRAIL were detected in fibrotic liver tissues. The results of TUNEL and α-SMA double-staining showed an increase in the apoptosis of activated HSCs during the spontaneous recovery phase. The knockdown of Smad2 reduced TRAIL-induced apoptosis in TGF-β1-activated human LX-2 cells and resulted in an increased expression of α-SMA and collagen I (Col. I). In contrast, the overexpression of Smad2 increased TRAIL-induced HSC apoptosis and reduced the expression of α-SMA and Col. I. The mechanisms underlying these findings were associated with the Smad2-mediated down-regulation of X-linked inhibitor of apoptosis protein (XIAP), resulting in enhanced caspase-3 activity and apoptosis. In conclusion, Smad2 enhances TRAIL-induced apoptosis in activated HSCs, which facilitates the resolution of hepatic fibrosis.

Xu F ，Zhou D ，Meng X ，Wang X ，Liu C ，Huang C ，Li J ，Zhang L ... -  《-》

NLRC5 regulates TGF-β1-induced proliferation and activation of hepatic stellate cells during hepatic fibrosis.

Therapeutic management of liver fibrosis remains an unsolved clinical problem. Hepatic accumulation of extracellular matrix, mainly collagen, is mediated by the production of transforming growth factor-β1 (TGF-β1) in hepatic stellate cells (HSCs). NLRC5, the largest member of the NLR protein family, has recently been identified as a critical regulator of immune responses. Novel evidence shows that NLRC5 is an important negative modulator of inflammatory pathways. Herein, we determined the regulation of NLRC5 in liver fibrogenesis and its underlying mechanisms. We have shown that NLRC5 was upregulated in human liver fibrotic tissues. Overexpression of NLRC5 resulted in an upregulation of collagen 1 and α-smooth muscle actin expression in HSC LX-2 cells, which was inhibited by NLRC5 knockdown with its siRNA. Furthermore, NLRC5 deficiency significantly suppressed TGF-β1-induced proliferation but increased apoptosis (i.e., increased caspases-3, DR4 and DR5) in LX-2 cells. In addition, knockdown of NLRC5 promoted the activation of NF-κB signaling pathways but abrogated phosphorylation of Smad2 and Smad3 proteins in response to TGF-β1. These results indicate that NLRC5 is a potent pro-fibrogenic molecule for HSC activation through TGF-β1/Smad and NF-κB signaling pathways. NLRC5 inhibition would be a promising therapeutic avenue for treating hepatic fibrosis.

Xu T ，Ni MM ，Xing-Li ，Li XF ，Meng XM ，Huang C ，Li J ... -  《-》

Knockdown of LOXL1 inhibits TGF-β1-induced proliferation and fibrogenesis of hepatic stellate cells by inhibition of Smad2/3 phosphorylation.

Liver fibrosis is pathological condition that seriously threatens human health. The lysyl oxidase (LOX) family has been reported to promote liver fibrosis. However, the effect of LOX-like 1 (LOXL1), a member of LOX family, on fibrogenesis of hepatic stellate cells (HSCs) remains unknown. The current study aimed to investigate the role of LOXL1 in liver fibrosis and the potential mechanism. We found that the mRNA and protein levels of LOXL1 were increased in transforming growth factor-beta 1 (TGF-β1)-stimulated human hepatic stellate cell line LX-2. Knockdown of LOXL1 inhibited the proliferation of TGF-β1-stimulated LX-2 cells. Knockdown of LOXL1 suppressed TGF-β1-induced expression of metalloproteinase type 1 (TIMP1), α-smooth muscle actin (α-SMA), and collagen type I (Col-I), as well as phosphorylation of Smad2 and Smad3 in LX-2 cells. In addition, the cell proliferation and fibrogenesis mediated by TGF-β1 stimulation and LOXL1 overexpression were abolished by knockdown of Smad2 and Smad3. Collectively, knockdown of LOXL1 suppressed cell proliferation and fibrogenesis in TGF-β1-stimulated HSCs via regulating the phosphorylation of Smad2/3.

Ma L ，Zeng Y ，Wei J ，Yang D ，Ding G ，Liu J ，Shang J ，Kang Y ，Ji X ... -  《-》

Newcastle disease virus represses the activation of human hepatic stellate cells and reverses the development of hepatic fibrosis in mice.

Li YL ，Wu J ，Wei D ，Zhang DW ，Feng H ，Chen ZN ，Bian H ... -  《-》

Fibroblast growth factor 21 attenuates hepatic fibrogenesis through TGF-β/smad2/3 and NF-κB signaling pathways.

Fibroblast growth factor 21 (FGF-21) is a secreted protein, which has anti-diabetic and lipocaic effects, but its ability to protect against hepatic fibrosis has not been studied. In this study, we investigated the ability of FGF-21 to attenuate dimethylnitrosamine (DMN)-induced hepatic fibrogenesis in mice and the mechanism of its action. Hepatic fibrosis was induced by injection of DMN, FGF-21 was administered to the mice once daily in association with DMN injection till the end of the experiment. Histopathological examination, tissue 4-hydroxyproline content and expressions of smooth muscle α-actin (α-SMA) and collagen I were measured to assess hepatic fibrosis. Ethanol/PDGF-BB-activated hepatic stellate cells (HSCs) were used to understand the mechanisms of FGF-21 inhibited hepatic fibrogenesis. Results showed that FGF-21 treatment attenuated hepatic fibrogenesis and was associated with a significant decrease in intrahepatic fibrogenesis, 4-hydroxyproline accumulation, α-SMA expression and collagen I deposition. FGF-21 treatment inhibited the activation of HSCs via down-regulating the expression of TGF-β, NF-κB nuclear translocation, phosphorylation levels of smad2/3 and IκBα. Besides, FGF-21 treatment caused activated HSC apoptosis with increasing expression of Caspase-3, and decreased the ratio of Bcl-2 to Bax. In conclusion, FGF-21 attenuates hepatic fibrogenesis and inhibits the activation of HSC warranting the use of FGF-21 as a potential therapeutic agent in the treatment of hepatic fibrosis.

Xu P ，Zhang Y ，Liu Y ，Yuan Q ，Song L ，Liu M ，Liu Z ，Yang Y ，Li J ，Li D ，Ren G ... -  《-》