Li Qiu-Hong, Shan Wan-Ting, Ju Ai-Xia, Zhao Jiao
School of Pharmacy, Heilongjiang University of Chinese Medicine Harbin 150040, China.
Zhongguo Zhong Yao Za Zhi. 2020 Aug;45(16):3961-3966. doi: 10.19540/j.cnki.cjcmm.20200512.202.
The enzymes CYP1 A2 and CYP3 A4 were measured by building a "Cocktail" probe drug and the incubation system of liver microsomes. The compatibility of Aconiti Lateralis Radix Praeparata combined with dried Rehmanniae Radix on CYP450 enzyme protein and gene expression was explored from the level of protein and molecular biology. It explored the molecular mechanism of compatibility detoxication of Aconiti Lateralis Radix Praeparata to provide scientific support for clinical safe and effective application of Aconiti Lateralis Radix Praeparata. The CYP450 enzyme activity was determined by using "Cocktail" probe drugs. The content of CYP450 enzyme was measured by CO reduction of differential spectrum method. The mRNA expression of CYP1 A2 and CYP3 A4 enzyme was detected by RT-PCR technology. Compared with the blank group, the CYP1 A2 and CYP3 A4 enzyme activity and mRNA expression were increased in the dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata group with significant differences(P<0.05), while the CYP3 A4 enzyme activity and mRNA expression were no influence in the Aconiti Lateralis Radix Praeparata group. The CYP3 A4 enzyme activity and mRNA expression were increased in the dried Rehmanniae Radix and the dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata group, and there were significant differences(P<0.05). The content of CYP450 enzyme was decreased in the Aconiti Lateralis Radix Praeparata group, contributed to extremely significant difference(P<0.01). The content of CYP450 enzyme was increased in the dried Rehmanniae Radix and the dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata group, and there were significant differences(P<0.05). The CYP1 A2 and CYP3 A4 enzyme activity and gene expression were enhanced after dried Rehmanniae Radix combined with Aconiti Lateralis Radix Praeparata. The metabolism of toxic ingredients of Aconiti Lateralis Radix Praeparata was accelerated to reach an effect of detoxication. The detoxication mechanism of compatibility of Aconiti Lateralis Radix Praeparata was verified from the viewpoint of liver metabolic enzymes.
采用构建“鸡尾酒”探针药物及肝微粒体孵育体系的方法测定CYP1 A2和CYP3 A4酶。从蛋白质和分子生物学水平探讨附子与地黄配伍对CYP450酶蛋白及基因表达的影响。探究附子配伍减毒的分子机制,为附子临床安全有效应用提供科学依据。采用“鸡尾酒”探针药物法测定CYP450酶活性,用差示光谱法CO还原法测定CYP450酶含量,运用RT-PCR技术检测CYP1 A2和CYP3 A4酶的mRNA表达。与空白组比较,地黄配伍附子组CYP1 A2和CYP3 A4酶活性及mRNA表达升高,差异有统计学意义(P<0.05),而附子组CYP3 A4酶活性及mRNA表达无影响。地黄组及地黄配伍附子组CYP3 A4酶活性及mRNA表达升高,差异有统计学意义(P<0.05)。附子组CYP450酶含量降低,差异极显著(P<0.01)。地黄组及地黄配伍附子组CYP450酶含量升高,差异有统计学意义(P<0.05)。地黄配伍附子后CYP1 A2和CYP3 A4酶活性及基因表达增强,加速了附子毒性成分的代谢,达到减毒效果。从肝脏代谢酶角度验证了附子配伍的减毒机制。