含砷、汞类矿物中药对小鼠肝/肠Cyp3a和肠道P-gp的影响

Effects of Mineral Chinese Medicine Containing Arsenic and Mercury on Mice Hepatic/Intestinal Cyp3a and Intestinal P-gp

目的研究雄黄、水飞雄黄、朱砂、氯化汞(Hg Cl2)对小鼠肝/肠细胞色素P450(cytochrome P450,CYP450)及肠道P-糖蛋白(P-glycoprotein,P-gp)的影响。方法将120只NIH小鼠随机分为纯水组,羧甲基纤维素钠(CMC)对照组,利福平组(40 mg·kg-1),酮康唑组(1.8 mg·kg-1),雄黄高、低剂量组(60,15 mg·kg-1),水飞雄黄高、低剂量组(60,15 mg·kg-1),朱砂高、低剂量组(120,30 mg·kg-1)和Hg Cl2高、低剂量组(0.2,0.05mg·kg-1)。小鼠每天灌胃2次,连续5 d。第6天灌胃1 h后摘取肝脏和全段小肠。用超微量ATP酶测试盒测定肠匀浆液中P-gp依赖性ATP酶的活性;用紫外分光光度法测定梯度离心分离的小鼠肝/肠微粒体中Cyp3a的活性;用实时定量PCR(real-time PCR,RT-PCR)法测定肝中Cyp3a11和肠道P-gp基因(Abcb1b)m RNA的表达。结果小鼠肠道P-gp偶联的ATP酶活性测定结果显示,雄黄、水飞雄黄、朱砂和Hg Cl2的高、低剂量组与CMC对照组比较差异均有统计学意义(P<0.05,P<0.01);用红霉素和氨基比林为探针药检测肝脏和小肠微粒体Cyp3a酶活性的结果显示,各供试物组的Cyp3a酶活性均显著高于CMC对照组(P<0.05,P<0.01),而RT-PCR结果显示各供试物均能显著诱导Cyp3a11和抑制Abcb1b的m RNA表达。结论雄黄、水飞雄黄、朱砂、Hg Cl2对小鼠肝脏和肠道中的Cyp3a均具有显著的诱导作用,而对肠道P-gp的转运活性则有显著抑制作用。

Objective To investigate the effects of realgar,levigated realgar,cinnabar and mercuric chloride（HgCl2） on mice hepatic/intestinal cytochrome P450 （CYP450）and intestinal P-glycoprotein（P-gp）. Methods One hundred and twenty NIH mice were randomly divided into normal group, sodium carboxymethylcellulose（SCMC） group, rifampicin group （40 mg·kg^-1）,ketoconazole group （1.8 mg·kg^-1）,high- and low-dose realgar groups（60,15 mg· kg^-1）,high- and low-dose levigated realgar groups （60,15 mg·kg^-1）,high- and low-dose cinnabar groups （120,30 mg·kg^-1）, and high- and low-dose HgCl2 groups（0.2, 0.05 mg·kg^-1）. The mice were orally treated with the corresponding medicine twice a day for 5 continuous days. On the experiment day 6, the liver and intestine were quickly isolated one hour after intragastrical administration. The activities of P-gp coupled ATPase in intestinal homogenate were detected by minim ATP enzyme test kit. The activities of Cyp3a in liver/intestine were detected by ultraviolet spectrophotometry,and the expression of hepatic Cyp3a11/Abcb1b mRNA was estimated by real-time PCR. Results P-gp coupled ATPase activities were significantly improved in both high and low dosage groups of realgar, levigated realgar,cinnabar and HgCl2,and the differences were significant compared with SCMC group （P〈 0.05,P〈 0.01）. Hepatic and intestinal microsomal Cyp3a activities showed by erythromycin and aminophenazone probes were significantly increased,hepatic Cyp3a11 mRNA expression was enhanced and intestinal Abcb1b mRNA expression was inhibited in both high and low dosage groups of realgar, levigated realgar, cinnabar and HgCl2 compared with SCMC group （P〈0.05,P〈0.01）. Conclusion Realgar,levigated realgar,cinnabar and HgCl2 exert an effect on obviously inducing the expression of CYP3a in mice liver and intestine while inhibiting intestinal P-gp activities.