摘要
目的研究中药复方861对血管紧张素Ⅱ(AngⅡ)引起的肝星状细胞增殖、细胞内钙浓度及胶原合成的影响.方法体外实验对象为肝星状细胞系(HSC-T6).细胞增殖采用甲基-3H胸腺嘧啶核苷(3H-TdR)掺入法,细胞内钙浓度测定用Fluo-3/AM(乙酰氧甲基酯)标记后共聚焦显微镜下测定荧光光密度值.培养上清液中Ⅰ型胶原(CoL-Ⅰ)用酶联免疫吸附(ELISA)法测定,Ⅲ型前胶原肽(PⅢP)的浓度用放免法检测.HSC-T6细胞CoL-Ⅰ及Ⅲ型胶原(CoL-Ⅲ)mRNA的水平用RT-PCR方法检测.结果 AngⅡ可促进HSC-T6细胞的增殖和细胞内钙浓度的增加,复方861可阻断这些变化.复方861也可降低HSC-T6细胞CoL-Ⅰ及PⅢP的分泌和CoL-Ⅰ及CoL-Ⅲ mRNA的水平.结论复方861通过作用于血管紧张素Ⅱ1型受体(AT1R)抑制肝星状细胞的增殖及胶原蛋白的合成,这可能是其抗纤维化的机理之一.
Objective To observe the effects of compound 861 (Cpd 861) on hepatic stellate cell line (HSC - T6) proliferation, intracellular calcium concentration and collagen synthesis caused by Angll . Methods Cell proliferation of HSC - T6 was detected with Methyl - 3H thymidine (3H - TdR) incorporation. Intracellular calcium, loaded with Fluo - 3 acetoxymethyl ester, was measured with laser scanning confocal microscope in HSC - T6. Collagen synthesis and gene expression in HSC - T6 were detected by enzyme linked immunosorbent assay (ELISA) and RT- PCR. Results AngII could cause an increase in intracellular calcium concentration and cell proliferation. All these effects were blocked by Cpd 861. Cpd 861 downregulated the elevation of Col I and Col III mRNA levels in HSC - T6, and decreased Co I and PIII P levels in culture supernants of HSC - T6. Conclusion Cpd 861 can decrease cell proliferation, intracellular calcium concentration and cell collagen synthesis via AT1R.
出处
《临床和实验医学杂志》
2003年第3期145-148,共4页
Journal of Clinical and Experimental Medicine
