去甲肾上腺素和酚妥拉明对大鼠肝星状细胞的影响  

Effects of noradrenaline and phentolamine in the rat hepatic stellate cells

作　　者：陈猛[1,2] 马勇[1,2] 魏伟[1] 李响[1] 张林[3] 

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药理学安徽省重点实验室,抗炎免疫药物安徽省工程技术研究中心,安徽省中药研究与开发重点实验室,合肥230032 [2]中国人民解放军第123中心医院,安徽蚌埠233015 [3]安徽医科大学公共卫生学院流行病与卫生统计学系,合肥230032

出　　处：《中国新药杂志》2009年第3期257-261,共5页Chinese Journal of New Drugs

基　　金：高等学校博士学科点专项科研基金(20060366003);安徽高校首批科技创新团队基金(2005TD003);安徽省研究实验基地优秀中青年科研带头人基金(200622805)

摘　　要：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 determine the effects of a-adrenoceptor agonist （noradrenaline, NA） and antagonist （phentolamine, PTL） in the rat hepatic stellate cells. Methods： After HSC-T6 cells were recovered and cultured, their proliferation was evaluated by MTT colorimetric assay. The levels of HA and PCⅢ released from the cultured hepatic stellate cells （HSC） was determined by radioimmunoassay （RIA） ; the apoptosis of HSC-T6 cells was determined by flow cytometric analysis; the expression of TIMP-1 and MMP-13 proteins was detected by immunocytochemical technique. Results： In the in vitro experiments, NA（ 10^-5, 10^-7 and 10^-9 mol·L^-1 ） promoted the proliferation of HSC-T6 cells, increased the expression of TIMP-1 protein and the levels of HA and PCⅢ released from the cultured HSC, and inhibited the apoptosis of HSC-T6 cells. Whereas, phentolamine （10^-5, 10^-7 and 10^-9 mol·L^-1） inhibited the proliferation of HSC-T6 cells, promoted the expression of MMP-13 protein, and induced the apoptosis of HSC-T6 cells. Conclusion： NA can promote the hepatic fibrosis, while PTL has protective effect on hepatic fibrosis. The mechanism might relate to the proliferation, apoptosis, and the release of HA and PCⅢ of HSC-T6 cells.

关 键 词：凋亡 肝星状细胞 Α-肾上腺素受体 肝纤维化 

分 类 号：R971.9[医药卫生—药品] R965.1[医药卫生—药学]