热毒宁注射液对大鼠肝微粒体CYP450酶的诱导作用研究

Study on Induction of Rat Liver Microsomes Cytochrome P450 Enzymes by Re-Du-Ning Injection

目的:研究热毒宁注射液(RDN)对大鼠肝微粒体CYP450酶的诱导作用。方法:将SD大鼠随机分为空白溶剂组、阳性对照组和热毒宁注射液低、中、高剂量组(1、2、4 m L·kg-1·d-1),分别于连续给药7天后处死,取肝组织称重并制备肝微粒体。计算大鼠肝脏的脏器系数,以BCA法测定微粒体的蛋白含量,以"cocktail"法测定大鼠肝微粒体中5种重要的CYP450酶亚型CYP1A2、CYP2C9、CYP2C19、CYP2D6和CYP3A1/2的活性。结果:与空白溶剂组比较,阳性对照组大鼠的肝脏脏器系数、微粒体得率及各CYP450酶亚型的活性均明显增加(P<0.01)。热毒宁注射液低、中剂量组的脏器系数、微粒体得率和蛋白含量无显著性差异,但高剂量组的脏器系数和微粒体得率有显著性差异(P<0.05)。对酶活性的影响方面,热毒宁注射液能显著诱导大鼠CYP1A2的活性,且呈剂量依赖性;中、高剂量组对CYP2C9和CYP2C19均有一定的诱导作用;对CYP3A1/2和CYP2D6的作用不明显。结论:阳性对照组对大鼠肝微粒体CYP450酶有明显的诱导作用,所建立的实验体系可用于诱导作用的评价;热毒宁注射液对大鼠CYP1A2、CYP2C9和CYP2C19亚型有一定的诱导作用,对CYP3A1/2和CYP2D6无明显作用。

This study was aimed to investigate the induction effect ofRe-Du-Ning （RDN） Injection on rat liver microsome CYP450 enzymes. SD rats were randomly divided into the solvent control group, positive control group as well as the low, middle and high dose group of RDN （1, 2, 4 mL·kg-1·d-1）. After drugs were administrated continuously for 7 days, the rats were sacrificed. The liver was weighed and prepared to microsomes. Meanwhile, the liver coefficients of rats were calculated. And the protein content was detected by BCA method. Finally, activities of five important subtypes of CYP450 enzymes such as CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A1/2 were measured by the“cocktail” method. The results showed that the levels of liver coefficients, microsome yield rate and activities of CYP450 subtypes increased significantly in the positive control group compared with the solvent control group （P ＆lt; 0.01）. There was no significant difference on the levels of liver coefficients, microsome yield and protein content between the low and middle dose group of RDN. However, there was significant difference on the levels of liver coefficients and microsome yield in the high dose group （P ＆lt; 0.05）. In terms of the influence on enzyme activity, RDN Injection can significantly induce the activities of CYP1A2 with dose dependence. It can induce the activities of CYP2C9 and CYP2C19 at the middle and high dose. However, there was no obvious influence on the activities of CYP3A1/2 and CYP2D6. It was concluded that the positive control group can obviously induce activities of CYP450, which can be used in the evaluation of induction experiments. RDN Injection had induction effect on CYP1A2, CYP2C9 and CYP2C19. But it had no influence on the activities of CYP3A1/2 and CYP2D6.