去甲肾上腺素对体外肝星状细胞系凋亡的影响

Effects of norepinephrine on hepatic stellate cells apoptosis in vitro

目的探讨交感神经递质去甲肾上腺素(NE)对体外培养的肝星状细胞(HSCs)凋亡的影响。方法体外培养HSCs,用四甲基偶氮唑盐(MTT)法检测NE对HSCs增殖的影响;原位杂交凋亡检测(TUNEL)法观察NE及各受体亚型的阻滞剂对HSCs凋亡的影响;流式细胞术检测凋亡率。结果(1)不同浓度NE均促进HSCs增殖,并呈时间依赖性;NE浓度为10μmol/L时促增殖作用最显著;(2)10μmol/L浓度NE作用于HSCs24h,TUNEL和流式细胞术检测HSCs凋亡率均显著低于对照组(P<0.05);(3)加入各肾上腺素受体(AR)阻滞剂后HSCs凋亡率升高,其中α-AR和β2-AR阻滞剂作用最显著。结论交感神经递质NE对体外活化的HSCs具有促增殖作用,并可以抑制HSCs凋亡,主要是通过α-AR和β2-AR起作用的。

Objective To investigate the effects of NE on hepatic stellate cells （HSCs） apoptosis. Methods After NE and the antagonists of adrenoceptor subtypes were administered to HSCs, MTT method was used to evaluate cell proliferation. TUNEL assay and flow cytometry（FCM） were used to identify cell apoptosis. Results （ 1 ） Compared with control group, NE at concentrations of 1,10 and 100 μmol/L induced HSCs proliferation in a time-dependent manner; The action of NE at concentrations of 10μ mol/L reached the peak （0. 262 ±0. 085 vs 0. 048 ±0. 055, P 〈 0. 05）. （2）After exposure of HSCs to NE at concentration of 10μmol/L for 24 h, apoptosis rates decreased significantly as compared with control group（6. 60 ±3.05 vs 12. 60 ±4. 76 by TUNEL, P 〈0. 05; 2. 29 ±0. 22 vs 3.06 ± 0. 57 by FCM, P 〈 0. 05）. （ 3 ） The antagonists of adrenoceptor subtypes, especially of α- and β2- adrenoceptor, reversed the apoptosis greatly. Conclusion Sympathetic neurotransmitter, NE, could induce HSCs proliferation and inhibit HSCs apoptosis, mainly through α- and β2- adrenoreceptor.