氧化苦参碱对免疫性肝纤维化的胶原纤维合成和TGF-β_1mRNA的影响

Effect of Oxymatrine on Collagen Synthesis and Expression of TGF-β1 in an Immune Hepatic Fibrosis Model

目的:研究氧化苦参碱对小鼠免疫性肝纤维化胶原纤维合成和TGF-β_1 mRNA的影响。方法:将BABL/c小鼠随机分成正常对照组、阳性对照组、氧化苦参碱小剂量、大剂量治疗组,阳性对照组和氧化苦参碱治疗组每周静脉注射1次刀豆蛋白A,共20W。氧化苦参碱治疗组分别腹腔注射氧化苦参碱30mg/kg、60mg/kg。共20W,分别于注射刀豆蛋白A后5、12和20W取血低温保存,并每次处死1批小鼠取肝脏组织液氮低温保存,常规HE染色、VanGieson胶原纤维染色,图像分析系统定量分析肝内纤维组织含量。抽提小鼠肝脏组织总RNA,RT—PCR方法扩增检测肝组织内TGF-β_1、β-actin。凝胶图像分析系统扫描扩增产物的灰度值,以β-actin作为内参照,计算相对表达量。结果:氧化苦参碱治疗组血清ALT含量及肝组织内炎症活动度、纤维含量均低于阳性对照组,而且呈剂量依赖性,TGF-β_1 mRNA相对表达量下降。结论:氧化苦参碱有减轻小鼠肝脏炎症活动度并可能通过下调TGF-β_1的表达而抑制小鼠免疫性肝纤维化的发生。

To elucidate the effect of Oxymatrine on collagen synthesis and expression of TGF-β1 in an immune hepatic fibrosis model induced by Concanavalin A in BALB/c mice. Methods. Female BALB/c mice were divided into 4 groups,which were normal control group,positive control group and Oxymatrine-treated groups respectively. Positive control group and Oxymatrine-treated groups received weekly injections of Concanavalin A by intravenous injection (via the retro-orbital venous plexus under anesthesia). Oxymatrine-treated groups were administered intraperitoneal injections of Oxymatrine (30mg/kg of 60mg/kg, qd, ×20week). Normal control group mice were administered Saline. Blood were collected by venous puncture from mice at 5, 12 and 20 week after Concanavalin A administration and sera were stored under-30Cuntil they were measured. The livers of different groups were fixed in 10% Formalin for routine light microscopy. Fibrosis was assessed with Van Gieson staining and the fibrous tissue was measured with MPIAS 500 imaging system. Extracting total RNA from liver tissues and analysis transforming factor β1 messenger RNA by reverse transcription polymerase chain reaction. PCR products were electrophoresed on an ethidium bromide and visualized using ultraviolet light. Densitometric RT-PCR data were standardized withβ-actin signals. Results: The levels of serum ALT and expression of TGF-β1 were lower in the Oxymatrinetreated mice than the positive control group, and so were the degrees of inflammatory activity and fibrogenesis in liver tissues. Conclusion: The results indicated that Oxymatrine can ameliorate the inflammatory activity and fibrogenesis through downregulation the expression of TGFβ1 mRNA in chronic T-cell-mediated immune hepatic fibrosis mouse models induced by Concanavalin A.