丹参酮Ⅱ_A磺酸钠治疗先天性人巨细胞病毒性肝炎小鼠的实验研究

Experimental study of tanshinone A sulfonic acid sodium in the treatment of congenital cytomegalovirus hepatitis in mice

目的建立小鼠先天性人巨细胞病毒肝炎动物模型,研究丹参酮Ⅱ_A磺酸钠的治疗机制,为治疗人类感染先天性巨细胞病毒肝炎的合理用药提供理论依据。方法 Balb/c小鼠80只(雌雄各半),配对后随机分对照组、病毒感染组、丹参酮大剂量组、丹参酮小剂量组,丹参酮大、小剂量组经腹腔注射丹参酮Ⅱ_A磺酸钠1、0.5 mg/(10 g·d),对照组、病毒感染组注射等体积细胞维持液DMEM,四组均治疗21d。放射免疫法检测治疗前后小鼠血清肝纤维化血清学指标:层黏连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(cⅣ)、透明质酸(HA)、转化生长因子-β_1(TGF-β_1),小鼠肝功能:总胆红素(STB)、胆汁酸(TBA)、丙氨酸氨基转移酶(ALT)、谷氨酰转移酶(GGT)、天冬氨酸氨基转移酶(AST),肝微粒体细胞色素P450亚型酶:CYP2E1、CYP1A2、CYP3A4、CYP2D6、CYP2C9活性。结果经丹参酮Ⅱ_A磺酸钠治疗21 d后,丹参酮大小剂量组LN、PC-Ⅲ、cⅣ、HA、TGF-β_1、STB、TBA、ALT、GGT、AST及CYP2E1、CYP3A4酶活性均有不同程度降低,与病毒感染组比较,差异均有统计学意义(P<0.05),丹参酮大、小剂量组组间比较,差异无统计学意义(P>0.05)。结论丹参酮Ⅱ_A磺酸钠对小鼠接种人巨细胞病毒性肝炎有明确的治疗作用,机制与其活血化瘀、降低TGF-β_1、抑制小鼠肝微粒体细胞色素_(450)亚型酶酶活性,促进肝功能恢复有关。

Objective To research the mechanisms of sodium tanshinone A sulfonate in the treatment of animal model of murine congenital human cytomegalovirus hepatitis and to provide a theoretical basis for the treatment of congenital human cytomegalovirus hepatitis.Methods Eighty Balb/c mice（male and female） were paired and randomly divided into control group,viral infection group,small dose of tanshinone group and high dose of tanshinone group.Mice of tanshinone group were injected sodium tanshinone A sulfonate[0.5 mg/（10 g·d）,1 mg/（10 g·d）]through intraperitone,and the mice of control group and infection group were injected DMEM.All the mice were treated for 21 d.The serum serologic indicators of hepatic fibrosis,such as laminin（LN） type Ⅲ procollagen（PC-Ⅲ）,type IV collagen（cⅣ）,hyaluronic acid（HA） and transforming growth factor-β_1（TGF-β_1）,were measured before and after treatment by Radioimmunoassay.The indexes of liver function of mice,such as,the content of total bilirubin（STB）,bile acid（TBA）,alanine aminotransferase（ALT）,glutamyl transferase（GGT）,aspartate aminotransferase（AST） and the activity of hepatic microsomal cytochrome P_（450） isoforms（CYP2E1,CYP1A2,CYP3A4,CYP2D6,CYP2C9） were defermined.Results The content of LN,PC-Ⅲ,cⅣ,HA,TGF-β_1,STB TBA,ALT,GGT,AST,and the activity of CYP2E1 and CYP3A4 in sodium tanshinone A sulfonic group were lower than those of viral infection group at 21 d after treatment（P〈0.05）.There was no significant difference between the two tanshinone groups（P〈0.05）.Conclusion Sodium tanshinone A sulfonate has definite therapeutic effect on mice inoculated with human cytomegalovirus hepatitis,and the mechanism is related to the decreased sodium tanshinone A sulfonate TGF-β_1 and liver microsomal cytochrome P_（450） isoform enzyme activity.