论文部分内容阅读
目的:探讨α-肾上腺素激动剂去甲肾上腺素和拮抗剂酚妥拉明对体外活化的肝星状细胞(HSC-T6)的影响。方法:体外对HSC-T6进行复苏和培养传代;MTT法检测HSC-T6的增殖;放射免疫法检测HSC-T6培养上清液中透明质酸(HA)和III型前胶原(PCIII)的浓度;流式细胞仪检测HSC-T6凋亡率;免疫细胞化学染色检测组织基质金属蛋白酶抑制因子-1(TIMP-1)和基质金属酶-13(MMP-13)蛋白的表达。结果:在体外培养的活化HSC-T6中,去甲肾上腺素(10-5,10-7,10-9mol.L-1)能促进细胞增殖,升高分泌HA和PCIII水平,增强TIMP-1蛋白的阳性表达并降低其凋亡率;酚妥拉明(10-5,10-7,10-9mol.L-1)能抑制细胞增殖,促进MMP-13的阳性表达并增加HSC-T6的凋亡率。结论:去甲肾上腺素具有促进肝纤维化的作用,酚妥拉明则具有抗纤维化的作用,该作用与其对HSC-T6增殖、凋亡及HA和PCIII分泌水平调控有关。
Objective: To investigate the effects of α-adrenergic agonist norepinephrine and antagonist phentolamine on activated hepatic stellate cells (HSC-T6) in vitro. Methods: HSC-T6 was resuscitated and cultured in vitro. The proliferation of HSC-T6 was detected by MTT assay. The concentrations of hyaluronic acid (HA) and type III procollagen (PCIII) in HSC-T6 culture supernatant were measured by radioimmunoassay The apoptosis rate of HSC-T6 was detected by flow cytometry. The expressions of TIMP-1 and MMP-13 were detected by immunocytochemistry. Results: In activated HSC-T6 cultured in vitro, norepinephrine (10-5, 10-7, 10-9mol.L-1) can promote cell proliferation, increase secretion of HA and PCIII, and enhance the expression of TIMP-1 Phentolamine (10-5, 10-7, 10-9mol.L-1) could inhibit cell proliferation, promote the positive expression of MMP-13 and increase the expression of HSC-T6 Apoptosis rate. Conclusion: Norepinephrine can promote hepatic fibrosis and phentolamine has anti-fibrosis effect, which is related to the regulation of HSC-T6 proliferation, apoptosis and the secretion of HA and PCIII.