褪黑素对CCl_4诱导大鼠肝纤维化模型肝星状细胞凋亡和功能的影响

The influence of melatonin on the apoptosis and function of hepatic stellate cells from the rat hepatic fibrosis model induced by carbon tetrarchloride

目的探讨褪黑素(MT)对肝星状细胞(HSC)凋亡和功能的影响。方法予SD大鼠背部皮下注射CCl4造肝纤维化模型。体内实验MT组大鼠灌胃给予MT〔0.125、0.5、2.0mg/(kg.d)〕8周,HE染色观察病理改变,放免法检测血清透明质酸(HA)与Ⅲ型前胶原(PCⅢ)。体外实验从模型组来源的HSC给予MT(10、0.1μmol/L和1nmol/L),从正常组来源的HSC给予生理盐水;48h后,流式细胞术检测HSC细胞凋亡,放免法检测培养上清中HA、PCⅢ。结果体内实验中,MT各剂量组均明显改善CCl4引起的肝脏病理改变,降低血清HA、PCⅢ。体外实验中,MT10μmol/L显著诱导HSC凋亡,各浓度MT均明显降低HA、PCⅢ的水平。结论MT可能通过诱导HSC凋亡和抑制其合成或分泌HA与PCⅢ而改善CCl4致大鼠肝纤维化。

Objective To explore the influence of melatonin（MT） on the apoptosis and function of hepatic stellate cells （HSC）. Methods The model of rat hepatic fibrosis was prepared by back subcutaneous injection of CCL4. In vivo,rats in MT groups were intragastric gavage administrated MT[0. 125,0. 5,2.0 mg/（ kg · d） ] for 8 weeks,the pathological changes were determined by HE stain, the levers of serum hyaluronic acid（HA） and procollagen type Ⅲ （PC Ⅲ ） were determinded by radioimmunoassay. In vitro, HSC isolated from model group rats was exposed to MT （ 10,0. 1 μmol/L and lnmol/L）, and HSC from normal group rats was exposed to NS. After 48 h,the apoptosis of HSC was determined by flow cytometry, and the levers of HA and PC III were determinded by radioimmunoassay. Results In vivo, liver pathological changes induced by CCl4 were relieved, and the levers of serum HA and PC III were reduced by MT with each dose. In vitro,the apoptosis of HSC were significantly induced by MT of 10 μmol/L, and the levers of HA and PC Ⅲ were reduced by MT of all concentrations. Conclusion Melatonin probably relieves rat hepatic fibrosis induced by CCl4 through inducing the apoptosis or inhibiting the synthesis or secretion of HA and PC Ⅲ of HSC.