蛴螬肽提取物对铅致小鼠肾中毒的解毒保护作用机制研究

Detoxification and protection mechanism of grub peptide extract on lead-induced nephrotoxicity in mice

目的本实验通过构建醋酸铅致小鼠肾中毒模型,探讨蛴螬肽提取物对小鼠的解毒保护作用机制,为机体铅性肾病的防治提供试验依据。方法随机取小鼠分为对照组、模型组、阳性药组以及不同剂量蛴螬肽组(80、160、320 mg/kg),除对照组外所有组小鼠隔天腹腔注射醋酸铅20 mg/kg,连续15 d。此时对照组和模型组小鼠同时灌服生理盐水而阳性药组小鼠灌服二巯基丁二酸(DMSA 70 mg/kg)混悬液,蛴螬肽组小鼠灌服不同剂量蛴螬肽提取物,每天1次连续15 d,采用HE染色显微镜观察肾组织状态;检测肾功能指标(BUN、Cr)和肾组织中抗氧化酶水平(SOD、GSH-Px)及过氧化物(MDA)含量;RT-PCR、Western Blot技术检测分析Ⅱ相解毒酶(NQO1)、抗氧化酶(HO-1)和信号分子(Nrf2)基因和蛋白表达水平。结果蛴螬肽组与模型组小鼠相比,体重增加但低于对照组,肾组织形态显著改善,血清中BUN、Cr水平显著降低(P<0.05),肾组织中抗氧化酶(SOD、GSH-Px)水平显著提高(P<0.05),MDA含量显著减少,Ⅱ相解毒酶基因(NQO1)、抗氧化酶(HO-1)及信号分子(Nrf2)mRNA及蛋白表达水平均显著上调(P<0.01)。结论蛴螬肽提取物激活Nrf2-ARE信号通路,增强铅中毒小鼠的抗氧化功能和提高解毒酶基因表达而发挥其解毒保护作用。

Objective In this experiment,a mouse model of nephropathy induced by lead acetate was created to study the mechanism of detoxification and protection of grub peptide extract on mice,which provided experimental evidence for the prevention and treatment of lead-induced nephropathy.Methods Mice were randomly divided into a Control group,Model group,Positive drug group,and Grub peptide groups.The Grub peptide groups were given different doses(80,160,320 mg/kg)of grub peptide.All mice,except the Control group,were injected intraperitoneally with 20 mg/kg lead acetate every other day,for 15 consecutive days.The mice in the Control group and Model group were then fed normal saline,while the mice in the Positive drug group were fed a dimercaptosuccinic acid(DMSA 70 mg/kg)suspension.The mice in the Grub peptide groups were fed different doses of grub peptide extract.The state of the kidney tissue was observed by HE staining once a day for 15 days.The renal function indexes(BUN,Cr),antioxidant enzyme levels(SOD,GSHPx),and peroxide(MDA)content in renal tissue were detected.RT-PCR and Western Blot techniques were used to detect and analyze the gene and protein expression levels of phase Ⅱ detoxification enzyme(NQO1),an antioxidant enzyme(HO-1),and a signaling molecule(Nrf2).Results The weight of mice in the Grub peptide groups was greater than those in the Model group;however,they were less than those in the Control group,and the morphology of the kidney tissue was significantly better.Compared with the control group,the morphology of the kidney tissue of mice in the Grub peptide groups was significantly improved,the level of BUN and Cr in the serum was significantly less(P<0.05),the level of antioxidant enzymes(SOD,GSH-Px)in renal tissue was significantly less(P<0.05),and the content of MDA was significantly less.The phase Ⅱ detoxification enzyme gene(NQO1),antioxidant enzyme(HO-1),and signal molecule(Nrf2)mRNA,and protein expression levels were significantly up-regulated(P<0.01).Conclusions Grub peptides can activate Nrf2-ARE signaling pathway to enhance antioxidant function and increase the gene expression of detoxification enzymes in lead-poisoned mice.The protective detoxification effect can then be extracted.