归脾汤对雷公藤醇提物致肝损伤大鼠肝微粒体CYP3A4酶活性的影响

Influence of CYP3A4 Activity of Guipi Tang to Acute Liver Injury Induced by Extract from Tripterygium wilfordii by Ethanol

目的：建立一个快速、准确的测定大鼠肝微粒体中CYP3A4酶活性的HPLC,研究雷公藤醇提物对肝损伤大鼠肝微粒体CYP3A4酶活性的影响,进而探讨归脾汤对其保护机制。方法：50只大鼠随机分为正常组、模型组、归脾汤组、P450诱导组、P450抑制组。正常组、模型组灌服生理盐水,归脾汤组灌服归脾汤（9g·kg-1）,P450诱导组、P450抑制组分别腹腔注射地塞米松（100mg·kg-1）和酮康唑（80mg·kg-1）,连续ig 4d后,再以雷公藤醇提物（3.25mg·kg-1）ig 3d造模。选用HPLC以咪达唑仑为探针药物,检测各组大鼠肝微粒体中CYP3A4酶活性。结果：在所建立的HPLC条件下,咪达唑仑的出峰时间在8.960min,能够完全分离,且无其他生物基质峰干扰;线性范围是0.25-20μmol·L-1（r=0.9999）,符合检测要求,方法可靠。结果显示,与正常组比较,模型组大鼠CYP3A4活性明显降低,差异有统计学意义（P〈0.05）;与模型组比较,归脾汤组、CYP450诱导组大鼠CYP3A4活性亦明显升高,差异有统计学意义（P〈0.05）;归脾汤组与CYP450诱导组之间比较,大鼠CYP3A4活性差异不明显,无统计学意义。结论：本方法能够对CYP3A4活性进行准确评价,雷公藤对大鼠肝微粒体CYP3A4酶活性有一定的抑制作用,归脾汤可能通过诱导CYP3A4活性而降低雷公藤的毒性从而发挥对肝脏的保护作用。

Objective： The aim of this study was to establish a rapid and accurate method in order to evaluate the activity of CYP3A4 by the high-performance liquid chromatography（HPLC）.The activity of CYP3A4 was evaluated to explore the protective effect and mechanism of Guipi Tang on rats＇ acute liver injury which is resulted from the extract from Tripterygium wilfordii by Ethanol.Method： Totally 50 rats were randomly divided into control group,model group,Guipi Tang group,P450 induction group,and P450 inhibition group.The control group and the model group was perfused with physiological saline,the Guipi Tang group was perfused with Guipi Tang（9.0g·kg-1）.The P450 induction group and P450 inhibition group have received injection of DXM（100mg·kg-1） and KCZ（80mg·kg-1） in intraperitoneal respectively.After ig 4 days,and perfused with T.wilfordii（3.25g·kg-1） for 3 days,the model was established.The activity of CYP3A4 of rat liver microsome was measured by HPLC with midazolam as a probe drug.Result： The retention time of midazolam appeared at 8.960min under the condition of HPLC,while there were no interference peaks.Its suitable linear range was 0.25 - 20μmol·L-1（r = 0.999 9）,which met the requirement of the whole examination.Compared with the control group,the activity of CYP3A4 in model group decreased（P〈0.05）.Compared with the model group,Guipi Tang and P450 induction group increase the CYP3A4 activity（P〈0.05）.Conclusion： The evaluation method of CYP3A4 activity is accurate.T.wilfordii can inhibit CYP3A4 activity significantly.Guipi Tang could reduce the toxicity of T.wilfordii and protect liver through inducing the CYP3A4 activity.