Zhao Yumin, Dang Zhisheng, Xu Shuo, Chong Shigui
.
Biol Chem. 2017 Nov 27;398(12):1357-1366. doi: 10.1515/hsz-2017-0177.
The study aimed to explore the regulation of heat shock protein 47 (HSP47) on expressions of receptors associated with hepatic stellate cell (HSC) in liver fibrosis mouse models induced by Schistosoma japonicum (S. japonicum). Mouse fibroblasts (NIH/3T3) were transfected with HSP47 shRNA plasmid by lipofectamine transfection, and experimental fibrosis in HSCs was studied in S. japonicum mouse models treated with HSP47 shRNA in vivo. HSP47 expression was assessed using Western blot and real-time PCR. Flow cytometry was adopted to determine the expression of cell membrane receptors. HSP47-shRNA could markedly down-regulate the expression of collagen (Col1a1 and Col3a1). The expressions of HSP47, endothelin receptor A (ETAR) and endothelin receptor B (ETBR) significantly increased in the liver tissue of infected mice. However, the expressions of ETAR and HSP47 and ETBR remarkably decreased after the administration of HSP47 shRNA in vitro and in vivo. ETAR and ETBR levels were found to be positively correlated with HSP47 expression. HSP47 might exert influence on liver fibrosis via the regulation of ETAR and ETBR.
本研究旨在探讨热休克蛋白47(HSP47)对日本血吸虫(S. japonicum)诱导的肝纤维化小鼠模型中肝星状细胞(HSC)相关受体表达的调控作用。通过脂质体转染将HSP47 shRNA质粒转染至小鼠成纤维细胞(NIH/3T3),并在体内用HSP47 shRNA处理的日本血吸虫小鼠模型中研究HSC中的实验性纤维化。采用蛋白质免疫印迹法和实时荧光定量PCR评估HSP47的表达。采用流式细胞术测定细胞膜受体的表达。HSP47-shRNA可显著下调胶原蛋白(Col1a1和Col3a1)的表达。感染小鼠肝脏组织中HSP47、内皮素受体A(ETAR)和内皮素受体B(ETBR)的表达显著增加。然而,在体外和体内给予HSP47 shRNA后,ETAR、HSP47和ETBR的表达显著降低。发现ETAR和ETBR水平与HSP47表达呈正相关。HSP47可能通过调控ETAR和ETBR对肝纤维化产生影响。