摘要
目的观察肝细胞对原代肝星状细胞(HSC)增殖、表型、胶原表达等生物学特性的影响,并为探讨其作用机制提供线索。方法原位灌流分离大鼠HSC,与大鼠肝细胞系BRL共培养48 h。在相差显微镜下观察HSC的表型变化,用Western blot方法检测其α-平滑肌肌动蛋白(α-SMA)、Ⅰ型前胶原的表达,TUNEL方法检测其凋亡。取BRL细胞的培养上清液培养原代HSC,用MTT法检测其增殖。采用抗体芯片技术显示BRL细胞的培养上清液与对照培养基之间细胞因子的表达差异。结果BRL细胞的培养上清液明显促进HSC增殖(P<0.01)。与BRL细胞共培养的原代HSC胞体收缩性增强,Ⅰ型前胶原及α-SMA的表达上调,凋亡增加(P<0.01)。较对照培养基,BRL细胞的培养上清液中血管内皮生长因子(VEGF)、单核细胞趋化蛋白-1(MCP-1)、金属蛋白酶组织抑制剂(TIMP-1)等三种细胞因子显著上调。结论肝细胞可促进原代HSC的增殖与活化,其机制可能与肝细胞分泌VEGF、MCP-1、TIMP-1等细胞因子有关。
Objective To observe the effect of hepatocytes on the biologic activity of primary hepatic stellate cells （HSC）, and to investigate its mechanism. Methods HSC isolated from SD rats were cocultured with rat hepatocyte line BRL for 48 h, then the phenotypic alternation of HSC was observed under phase contrast microscope. The expressions of α-smooth muscle actin（α-SMA） and precollagen type I were detected by Western blot. The apoptosis of HSC was detected by TUNEL method. The primary HSC was cultured in the supernatant of BRL cells or control culture medium for 48 h, and the proliferation of HSC was detected by MTT method. The differences of cytokines＇ expression between the supernatant of BRL cells and control culture medium were analyzed by using cytokines antibody arrays. Results BRL cells significantly facilitated proliferation, apoptosis, contractibility, α-SMA and procollagen type Ⅰ expression of primary HSC （P 〈 0.01 ）. Antibody arrays indicated that there was a higher up-regulation of vascular endothelial growth factor（VEGF） , monocyte chemoattractant protein-1 （ MCP-1 ） , and tissue inhibitor of metalloproteinase-1 （TIMP-1） in the supernatant of BRL cells than that in control culture medium. Conclusion Hepatocytes stimulate proliferation and activation of HSC, which may due to its secretion of VEGF, MCP-1 and TIMP-1.
出处
《胃肠病学和肝病学杂志》
CAS
2008年第11期933-936,共4页
Chinese Journal of Gastroenterology and Hepatology
关键词
肝细胞
肝星状细胞
增殖
胶原
细胞因子
Hepatocyte
Hepatic stellate cell
Proliferation
Collagen
Cytokine
