Li Yi, Wu Wei, Jiang Yong-Fang, Wang Kang-Kai
Institute of Hepatology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2007 Aug;32(4):650-5.
To determine the effect of heat shock protein 47 (HSP47) on the expression of collagen I induced by transforming growth factor beta(1) (TGF-beta(1)) in hepatic stellate cell-T6 (HSC-T6) cells.
We used 1 ng/mL and 10 ng/mL recombinant human TGF-beta(1) to stimulate the cultured HSC-T6 cells. Heat shock response (HSR) and antisense oligonucleotides of HSP47 were used to induce and block the expression of HSP47, respectively. The expressions of HSP47 and collagen I were detected by Western blot and the cell viability was observed by MTT assay.
Both HSP47 and collagen I were expressed in normal HSC-T6 cells. Collagen I and HSP47 expression could be induced by both 1 ng/mL and 10 ng/mL TGF-beta(1) and collagen I was expressed the most after the treatment with 10 ng/mL TGF-beta(1). Although HSR could not affect the synthesis of collagen I as it induced the HSP47 expression, HSR could promote the expression of collagen I induced by TGF-beta(1). With no effect on the cell viability, antisense oligonucleotides could significantly inhibit HSR-mediated HSP47 expression and TGF-beta(1)-induced collagen I synthesis.
Over-expression of HSP47 enhances TGF-beta(1)-induced expression of collagen I in HSC-T6 cells, and HSP47 may play important roles in the process of hepatic fibrosis.
确定热休克蛋白47(HSP47)对肝星状细胞-T6(HSC-T6)中转化生长因子β1(TGF-β1)诱导的I型胶原蛋白表达的影响。
我们使用1 ng/mL和10 ng/mL重组人TGF-β1刺激培养的HSC-T6细胞。热休克反应(HSR)和HSP47反义寡核苷酸分别用于诱导和阻断HSP47的表达。通过蛋白质免疫印迹法检测HSP47和I型胶原蛋白的表达,并通过MTT法观察细胞活力。
HSP47和I型胶原蛋白在正常HSC-T6细胞中均有表达。1 ng/mL和10 ng/mL TGF-β1均可诱导I型胶原蛋白和HSP47表达,且10 ng/mL TGF-β1处理后I型胶原蛋白表达量最高。虽然热休克反应在诱导HSP47表达时不影响I型胶原蛋白的合成,但可促进TGF-β1诱导的I型胶原蛋白表达。反义寡核苷酸对细胞活力无影响,但可显著抑制热休克反应介导的HSP47表达和TGF-β1诱导的I型胶原蛋白合成。
HSP47的过表达增强了TGF-β1诱导的HSC-T6细胞中I型胶原蛋白的表达,HSP47可能在肝纤维化过程中发挥重要作用。