槲皮素对17α-乙炔雌二醇诱导胆汁淤积的肝保护作用研究

Hepatoprotective effect of quercetin on 17α-ethynylestradiol-induced cholestasis in rats

目的探讨槲皮素(Que)对17α-乙炔雌二醇(EE)诱导胆汁淤积的肝保护作用及可能机制。方法采用随机数字表法将SD大鼠分为对照组、模型组、给药组(Que 50 mg/kg组和Que 100 mg/kg组),每组8只。模型组及给药组连续5 d皮下注射5 mg/kg EE造模,对照组皮下注射EE溶剂丙二醇。Que 50 mg/kg组和Que 100 mg/kg组分别予50 mg/kg和100 mg/kg Que(溶于0.5%羧甲基纤维素钠溶液)灌胃,对照组及模型组予Que溶剂0.5%羧甲基纤维素钠溶液灌胃。采用HE染色和血清生化指标评价大鼠肝损伤情况,按照胆汁酸测定试剂盒测定血清与肝脏中胆汁酸水平;眼科剪顺着胆管方向剪一个小口,收集1 h内的胆汁酸,并计算胆汁流量;RT-qPCR测定胆汁酸摄取转运体、外排转运体、胆汁酸合成酶及代谢酶mRNA表达情况。结果与模型组相比,Que显著减轻EE诱导的胆汁淤积肝损伤,减轻大鼠血清AST、ALT和ALP水平(均P<0.05),增加大鼠胆汁流量(均P<0.01),降低血清及肝脏胆汁酸水平(均P<0.05),显著增加外排转运体胆汁酸盐输出泵、多药耐药相关蛋白(MRP)2、MRP3、MRP4和胆汁酸代谢酶细胞色素P450酶(CYP)3A2、CYP2B10、硫酸基转移酶2A1的表达(均P<0.05),显著抑制胆汁酸摄取转运体钠离子-牛磺胆酸共转运蛋白和胆汁酸合成酶CYP7A1和CYP8B1表达(均P<0.05)。结论Que可通过调节胆汁酸转运体与酶的表达缓解肝脏胆汁酸累积,从而发挥保肝利胆的功效。

Objective To investigate the effect of quercetin(Que)on 17α-ethynylestradiol(EE)-induced cholestatic liver injury and the possible mechanism.Methods Thirty-two SD rats were randomly divided into control group,model group,Que 50 mg/kg group and Que 100 mg/kg groups with 8 rats in each group.The model group and the Que treatment groups were injected subcutaneously with EE(5 mg/kg)for 5 consecutive days,and the control group was injected with EE solvent(propylene glycol).Que 50 mg/kg group and Que 100 mg/kg group were given 50 mg/kg and 100 mg/kg Que(dissolved in 0.5%sodium carboxymethyl cellulose solution)by gavage,respectively.The control group and model group were given Que solvent 0.5%sodium carboxymethyl cellulose solution.HE staining and serum biochemical indicators were used to evaluate the liver injury in rats,and the bile acid levels in serum and liver were determined according to the bile acid determination kit.Ophthalmological scissors were used to cut a small mouth along the direction of the bile duct to collect bile acids within 1 h and calculate the bile flow.RT-qPCR was used to determine the mRNA expression of bile acid efflux transporters,uptake transporters,bile acid synthase and metabolic enzymes.Results Compared with the model group,Que significantly attenuated EE-induced cholestasis liver injury,and reduced the levels of serum AST,ALT,and ALP in rats(all P<0.05),increased bile flow rate(both P<0.01)and decreased serum and liver bile acid levels(all P<0.05).Que significantly increased the expression of efflux transporters BSEP,MRP2,MRP3,MRP4 and bile acid metabolizing enzymes CYP3A2,CYP2B10,SULT2A1(all P<0.05).Que significantly inhibited the expression of bile acid uptake transporter NTCP and bile acid synthetic enzymes CYP7A1 and CYP8B1(all P<0.05).Conclusion Quercetin can alleviate the accumulation of bile acid in liver by regulating the expression of bile acid transporters and enzymes,thus exerting the hepatoprotective effect and promoting bile flow.