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目的:通过检测大鼠肝功能指标和肝组织CYP2E1、CYP3A4的基因、蛋白表达,初步观察醋甘遂与炙甘草配伍对肝脏的损伤以及对肝药酶2个亚型的诱导作用。方法:将大鼠按体质量随机分为正常组、炙甘草组(免煎颗粒剂)、醋甘遂组(研末)、炙甘草-醋甘遂组、甘遂半夏汤组。其中正常组给予生理盐水灌胃,其余各组给予相应药液灌胃,连续给药14天。实验结束时,腹主动脉取血分离血清和血浆检测谷草转氨酶(AST)、谷丙转氨酶(ALT)、乳酸脱氢酶(LDH)含量;取肝组织一部分用福尔马林固定观察肝组织病理形态,一部分液氮冻存,采用RT-PCR和western-blot技术检测肝组织中CYP2E1、CYP3A4的基因、蛋白表达情况。结果:与正常组相比,各给药组ALT含量各组变化不明显,无统计学差异(P>0.05),AST含量甘遂组以及醋甘遂-炙甘草配伍组、甘遂半夏汤组较正常组降低,其中甘遂半夏汤组降低显著,有统计学差异(P<0.05),LDH含量各给药组均较正常组显著降低,有统计学差异(P<0.05,P<0.01)。与空白组比较,炙甘草组、醋甘遂组、醋甘遂-炙甘草组和甘遂半夏汤组CYP2E1基金和蛋白表达均有所升高,CYP3A4的基因和蛋白表达则有所下降,二者均具有统计学差异(P<0.01),其中以醋甘遂-炙甘草组升高或降低最为明显;与醋甘遂-炙甘草组比较,甘遂半夏汤组下调CYP2E1基金和蛋白表达,上调CYP3A4的基因和蛋白表达。结论:本实验从分子机制上证实了醋甘遂和炙甘草配伍有增毒效应,以复方的形式使用则能降低毒性,但在生化指标方面未表现出明显增毒作用,可能还存在其他配伍机制有待进一步研究,且需要配合病理状态的研究以确定是否有增效作用。
OBJECTIVE: To detect the hepatic function of rats and the expression of CYP2E1 and CYP3A4 gene and protein in liver tissue, and to observe the injury of liver and the inductions of two subtypes of hepatic drug through the combination of vinegrew root and Zhigancao. Methods: According to body weight, rats were randomly divided into normal group, Zhigancao group (decocted granule), vinegar and kansui group (Zhimo), Zhigancao - vinegar group, Gan Sui Banxia Tang group. The normal group was given normal saline gavage, and the remaining groups were given the corresponding liquid gavage, continuous administration for 14 days. At the end of the experiment, serum and plasma were taken from the abdominal aorta to detect the content of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH); part of the liver tissue was fixed with formalin to observe the pathological changes of liver Morphology, part of the liquid nitrogen frozen, using RT-PCR and western-blot detection of liver tissue CYP2E1, CYP3A4 gene and protein expression. Results: Compared with the normal group, there was no significant difference in the ALT content in each group (P> 0.05). The content of AST in the Gan-Sui group and the control group (P <0.05). The content of LDH in each group was significantly lower than that in the normal group (P <0.05, P <0.05), and the difference was statistically significant 0.01). Compared with the blank group, CYP2E1 expression and protein expression in Zhigancao, Jiakuishui, Zhuguansui - Zhigancao and Gansui Banxiaxutang groups were increased, but the gene and protein expression of CYP3A4 was decreased, (P <0.01). The most obvious increase or decrease was in Kampuchew-Zhigancao group. Compared with the vinegar-kansui-Zhigancao group, the Kansui Banxiaxutang group down-regulated CYP2E1 fund and protein Expression, upregulation of CYP3A4 gene and protein expression. Conclusion: This experiment confirmed from the molecular mechanism of vinegar kuishui and Zhigancao compatibility increased toxic effects, the use of the compound can reduce the toxicity of the form, but no significant increase in biochemical indicators of toxicity, there may be other compatibility Mechanisms to be further studied, and the need to cooperate with the study of the pathological state to determine whether there is a synergistic effect.