辣木叶及辣木籽对大鼠CYP450酶3种亚型基因表达影响

目的:研究辣木叶及辣木籽对大鼠肝脏CYP450亚型酶中mRNA及蛋白表达量的影响。方法:将SD大鼠随机分为空白组(0.5%羧甲基纤维素钠混悬溶液)、辣木叶高、中、低剂量组(分别为0.813 8,0.406 9,0.203 5g·kg^-1);辣木籽高、中、低剂量组(分别为1.067 4,0.533 7,0.266 9g·kg^-1)。灌胃给药,给药容量为10mL·kg^-1,2次/天,连续给药14d,取大鼠肝脏,采用实时荧光定量PCR(RT-qPCR)及蛋白免疫印迹(Western-blot)法检测大鼠肝脏中CYP2E1、CYP3A1及CYP1A2的mRNA及蛋白相对表达量。结果:辣木叶高剂量组对CYP2E1的mRNA表达有明显抑制作用(P<0.05);在(0.813 8±0.203 5)g·kg^-1剂量范围内,辣木叶对CYP1A2mRNA表达的抑制作用随给药剂量增加而增加(P<0.05)。本实验剂量范围内的辣木籽对3个酶的mRNA表达都有明显的抑制作用(P<0.05),其抑制效率CYP2E1>CYP3A1>CYP1A2。高剂量组辣木叶及辣木籽对CYP2E1蛋白表达都有明显抑制作用(P<0.01);中、低剂量组的辣木籽对CYP3A1及CYP1A2蛋白表达都有不同程度的抑制作用(P<0.05;P<0.05);中剂量辣木叶对CYP1A2蛋白表达抑制作用显著(P<0.01)。结论:辣木叶及辣木籽对大鼠肝脏中3个亚型酶的mRNA及蛋白表达都有不同程度的抑制作用。

Effect of Moringa leaves and seed on mRNA and protein expression of CYP450 enzymes in rats

OBJECTIVE To research the effect of Moringa oleifera Lam. leaves and seed on mRNA and protein expression levels of CYP2E1, CYP3A1 and CYP1A2 subtypes of CYP450 enzymes in rat liver. METHODS SD rats were randomly divided into the normal control group (treated with 0.5% sodium carboxymethyl cellulose suspension solution), Moringa oleifera Lam. leaves high dose, middle dose, low dose groups (doses were 0.813 8, 0.406 9, 0.203 5 g·kg^-1, respectively), Moringa oleifera Lam. seed high dose, middle dose, low dose groups (doses were 1.067 4, 0.533 7, 0.266 9 g·kg^-1, respectively). Doses were given by intragastric administration at 10 mL·kg^-1,bid. The liver of rats was taken after continuous administration for 14 days. The mRNA expression levels of CYP2E1, CYP3A1 and CYP1A2 subtypes in rat liver were detected by real-time quantitative PCR technology, and the protein expression level was detected by Western-blot. RESULTS The CYP2E1 mRNA expression was inhibited significantly by high dose group of Moringa oleifera Lam. leaves (P<0.05). Within the dose range of (0.813 8±0.203 5) g·kg^-1, the inhibitory effect of Moringa oleifera Lam. leaves on the expression of CYP1A2 mRNA increased with the concentration (P<0.05). The mRNA expressions of three enzymes were inhibited significantly in this dose range of Moringa oleifera Lam. seed (P<0.05), inhibitory efficiencies were ordered CYP2E1 > CYP3A1 > CYP1A2. The expression of CYP2E1 protein was inhibited by high dose groups of Moringa leaves and seed (P<0.05). The middle and low dose groups of Moringa oleifera Lam. seed had different inhibition effects on the expression of CYP3A1 and CYP1A2 proteins (P<0.05). The expression of CYP1A2 protein was inhibited in middle dose group of Moringa oleifera Lam. leaves(P<0.01). CONCLUSION The expressions of mRNA and protein of 3 isoforms in rat liver are inhibited by Moringa oleifera Lam.leaves and seed in different degrees.