Li Tao, Leng Xi-Sheng, Zhu Ji-Ye, Wang Gang
Department of Hepatobiliary Surgery, Peking University People's Hospital Xizhimen South Street, West District, Beijing 100044, P. R. China.
Int J Clin Exp Pathol. 2015 Nov 1;8(11):14574-9. eCollection 2015.
Hepatic stellate cells (HSCs) play an important role in liver fibrosis. This study investigates the expression of hedgehog in HSC and the role of hedgehog signaling on activation and collagen secretion of HSC. Liver ex vivo perfusion with collagenase IV and density gradient centrifugation were used to isolate HSC. Expression of hedgehog signaling components Ihh, Smo, Ptc, Gli2 and Gli3 in HSC were detected by RT-PCR. Hedgehog siRNA vectors targeting Ihh, Smo and Gli2 were constructed and transfected into HSC respectively. Suppression of hedgehog signaling were detected by SYBR Green fluorescence quantitative RT-PCR. Effects of hedgehog signaling inhibition on HSC activation and collagen I secretion were analyzed. Hedgehog signaling components Ihh, Smo, Ptc, Gli2 and Gli3 were expressed in HSC. siRNA vectors targeting Ihh, Smo and Gli2 were successfully constructed and decreased target gene expression. Suppression of hedgehog signaling significantly decreased the expression of α-SMA in HSC (P<0.01). Collagen type I secretion of HSC were also significantly decreased (P<0.01). In summary, HSC activation and collagen secretion can be regulated by hedgehog signaling. Hedgehog may play a role in the pathogenesis of liver fibrosis.
肝星状细胞（HSCs）在肝纤维化中起重要作用。本研究调查了刺猬信号通路在肝星状细胞中的表达以及刺猬信号通路对肝星状细胞激活和胶原分泌的作用。采用IV型胶原酶肝脏离体灌注和密度梯度离心法分离肝星状细胞。通过RT-PCR检测肝星状细胞中刺猬信号通路成分Ihh、Smo、Ptc、Gli2和Gli3的表达。构建靶向Ihh、Smo和Gli2的刺猬信号通路小干扰RNA（siRNA）载体，并分别转染到肝星状细胞中。通过SYBR Green荧光定量RT-PCR检测刺猬信号通路的抑制情况。分析刺猬信号通路抑制对肝星状细胞激活和I型胶原分泌的影响。刺猬信号通路成分Ihh、Smo、Ptc、Gli2和Gli3在肝星状细胞中表达。成功构建了靶向Ihh、Smo和Gli2的siRNA载体，并降低了靶基因的表达。抑制刺猬信号通路可显著降低肝星状细胞中α-SMA的表达（P<0.01）。肝星状细胞I型胶原的分泌也显著减少（P<0.01）。综上所述，刺猬信号通路可调节肝星状细胞的激活和胶原分泌。刺猬信号通路可能在肝纤维化的发病机制中起作用。
