茵陈蒿汤及其不同组方提取物对雌激素诱导胆汁淤积大鼠的治疗作用及机制研究

目的：基于肝细胞膜转运体多药耐药相关蛋白1~4（multidrug resistant associated protein 1~4，Mrp1~4）、Na⁺依赖性牛磺胆酸共转运多肽（Na⁺-taurocholate co-transporting polypeptide，Ntcp），探讨茵陈蒿汤及其不同组方提取物对胆汁淤积大鼠的治疗作用。方法： 30只Wistar雄性大鼠随机分为6组，即正常组、对照组、茵陈组、茵陈栀子组、茵陈大黄组、茵陈蒿汤组；采用皮下连续注射苯甲酸雌二醇（estradio benzoate，EB，5 mg·kg^-1·d^-1）5 d制备胆汁淤积大鼠模型。茵陈组、茵陈栀子组、茵陈大黄组、茵陈蒿汤组分别灌胃给予茵陈（0.43 g·kg^-1·d^-1）、茵陈栀子（1.08 g·kg^-1·d^-1）、茵陈大黄（1.09 g·kg^-1·d^-1）、茵陈蒿汤（1.69 g·kg^-1·d^-1）提取物14 d。干预结束后，测定各组大鼠胆汁流速，血清生化及肝脏病理，并采用Western blot检测肝组织中Mrp1~4和Ntcp的表达。结果：茵陈蒿汤及不同配伍组干预14 d后，胆汁流速均显著升高，血清生化与肝脏病理较对照组均有明显改善；与对照组相比，茵陈组、茵陈栀子组、茵陈大黄组及茵陈蒿汤组Mrp2、Mrp3、Ntcp的表达均升高；Mrp4的表达均降低；此外，茵陈栀子组Mrp1的表达降低。结论：茵陈蒿汤及其不同组方提取物对雌激素诱导的大鼠胆汁淤积均有治疗作用；其疗效可能与调控转运体Mrp1-4和Ntcp的表达相关，且不同组方提取物对Mrp1~4和Ntcp的调控作用不同。

The therapeutic effect and the underlying mechanism of Yinchenhao decoction and its different modified combinations on estrogen induced cholestasis rats

OBJECTIVE To explore the therapeutic effects and the underlying mechanism of Yinchenhao decoction and its different modified combinations on estrogen induced cholestasis rats based on the liver membrane transporter Ntcp and Mrp1-4.METHODS Male Wistar rats were randomly divided into six groups:normal group, control group, Yinchen group, Yinchen plus Zhizi group, Yinchen plus Dahuang group, and Yinchenhao decoction group. Cholestasis model rats were induced via subcutaneous injection of estradiol benzoate (EB, 5 mg·kg^-1·d^-1) for 5 days. The doses of Yinchen, Yinchen plus Zhizi, Yinchen plus Dahuang and Yinchenhao decoction were 0.43, 1.08, 1.09, and 1.69 g·kg^-1·d^-1, respectively. After drug treatment for 14 days, bile flow rate, serum biochemical parameters, liver pathohistologic examination, and the expression of Ntcp and Mrp1 4 were further investigated.RESULTS After the treatment of Yinchenhao decoction and its different modified combinations for 14 days, the bile flow rate was obviously increased, serum biochemical parameters and liver pathohistology were improved significantly. Compared with control group, the expressions of Mrp2, Mrp3 and Ntcp were significantly increased and the expression of Mrp4 was significantly decreased in Yinchen group, Yinchen plus Zhizi group, Yinchen plus Dahuang group, and Yinchenhao decoction group. In addition, the expression of Mrp1 was significantly decreased in Yinchen plus Zhizi group.CONCLUSION Yinchenhao decoction and its different modified combinations can improve the estrogen induced rat cholestasis. The underlying mechanism may correlate with their effect on regulating the expression of transporters responsible for the balance of bile acid salt and bilirubin.