Jung Gwon-Soo, Jeon Jae-Han, Choi Yeon-Kyung, Jang Se Young, Park Soo Young, Kim Mi-Kyung, Shin Eui-Cheol, Jeong Won-Il, Lee In-Kyu, Kang Yu Na, Park Keun-Gyu
Department of Internal Medicine, Kyungpook National University School of Medicine, Daegu, Korea.
Department of Internal Medicine, Keimyung University School of Medicine, Daegu, Korea.
Liver Int. 2015 Oct;35(10):2233-45. doi: 10.1111/liv.12871. Epub 2015 Jun 3.
BACKGROUND & AIMS: An atypical orphan nuclear receptor small heterodimer partner (SHP) is known to be regulated by AMP-activated protein kinase (AMPK). Both of them inhibit TGF-β and Smad signalling and exhibit antifibrotic activity in the liver. However, little is known about the protective effects of SHP and AMPK against hepatitis c virus (HCV)-induced hepatic fibrosis.
Levels of SHP, p-AMPK and fibrotic markers in HCV-infected human liver and in Huh-7.5 cells infected with HCV genotype 2a (JFH-1) were investigated. The effect of adenovirus-mediated overexpression of SHP (Ad-SHP) and AMPK activation via metformin and 5-amino-1-b-D-ribofuranosyl-imidazole-4-carboxamide (AICAR) on fibrotic gene expression was evaluated in HCV-infected cells. Finally, we examined the effect of Ad-SHP and AMPK activators on invasion and activation of LX2 human HSCs induced by conditioned media from HCV-infected hepatocyte (CM).
In HCV-infected human livers and Huh-7.5 cells infected with HCV, SHP mRNA and protein levels were diminished compared with controls, whereas profibrotic factors were increased. Pharmacological AMPK activation recovered SHP expression, and Ad-SHP inhibited HCV-induced fibrotic gene expression. This finding was accompanied by inhibition of HCV-stimulated nuclear factor-kappa B, an inducer of TGF-β. Moreover, CytoSelect invasion assay revealed that enhanced activity and invasiveness of hepatic stellate cells induced by CM.
These results demonstrate that overexpression of SHP and activation of AMPK reverses profibrogenic features of HCV-infected cells by decreasing TGF-β and fibrotic gene expression. These findings provide a rationale for SHP as a possible therapeutic target against HCV-induced hepatic fibrosis.
已知一种非典型孤儿核受体小异二聚体伴侣（SHP）受AMP激活蛋白激酶（AMPK）调控。二者均抑制转化生长因子-β（TGF-β）和Smad信号传导，并在肝脏中表现出抗纤维化活性。然而，关于SHP和AMPK对丙型肝炎病毒（HCV）诱导的肝纤维化的保护作用知之甚少。
研究了HCV感染的人肝脏以及感染HCV基因型2a（JFH-1）的Huh-7.5细胞中SHP、p-AMPK和纤维化标志物的水平。评估了腺病毒介导的SHP过表达（Ad-SHP）以及通过二甲双胍和5-氨基-1-β-D-呋喃核糖基咪唑-4-甲酰胺（AICAR）激活AMPK对HCV感染细胞中纤维化基因表达造成的影响。最后，我们检测了Ad-SHP和AMPK激活剂对HCV感染的肝细胞条件培养基（CM）诱导的LX2人肝星状细胞侵袭和激活的影响。
在HCV感染的人肝脏和感染HCV的Huh-7.5细胞中，与对照组相比，SHP mRNA和蛋白水平降低，而促纤维化因子增加。药理学上激活AMPK可恢复SHP表达，且Ad-SHP抑制HCV诱导的纤维化基因表达。这一发现伴随着对HCV刺激的核因子-κB（一种TGF-β诱导剂）的抑制。此外，细胞侵袭分析显示CM诱导肝星状细胞的活性和侵袭性增强。
这些结果表明，SHP的过表达和AMPK的激活通过降低TGF-β和纤维化基因表达来逆转HCV感染细胞的促纤维化特征。这些发现为SHP作为抗HCV诱导的肝纤维化的潜在治疗靶点提供了理论依据。
