肝豆汤改良方对TX小鼠脑组织ASM、Cer及p38 MAPK mRNA和蛋白表达的影响

Effects of Modified Gandou Decoction on the mRNA and protein expression of ASM, Cer and p38 MAPK in the brain of TX mouse

目的:探讨肝豆汤改良方(MGDD)对TX小鼠脑组织酸性鞘磷脂水解酶(ASM)、神经酰胺(Cer)及p38丝裂原活化蛋白激酶(p38 MAPK)m RNA和蛋白表达的影响。方法:将140只TX小鼠随机分成1、3、5月龄模型组,3、5月龄MGDD组及3、5月龄丁苯酞组,每组20只,同时选用相同月龄60只DL小鼠分别为1、3、5月龄正常对照组,每组20只,饲养环境及喂食食物相同,均从入组时开始给药,MGDD治疗组按24.5g·kg^(-1)·d^(-1)灌胃,丁苯酞组给以丁苯酞混悬液0.156g·kg^(-1)·d^(-1)灌胃,正常对照组及模型组灌胃100m L·kg^(-1)·d^(-1) 0.9%氯化钠溶液,1次/d,收集处理标本,采用RT-PCR、免疫组织化学法及Western blot技术检测脑组织内ASM、Cer及p38 MAPK m RNA及蛋白表达水平。结果:模型组小鼠脑组织ASM、Cer及p38 MAPK mRNA及蛋白表达程度伴随着月龄的增加,表达有增多趋势,MGDD治疗组较模型组可显著降低小鼠脑内ASM、Cer及p38 MAPK mRNA及蛋白表达水平(P<0.05,P<0.01),且随着疗程的延长,抑制更加显著。结论:MGDD可能通过下调小鼠脑组织神经酰胺信号通路中ASM、Cer、p38 MAPK mRNA及蛋白的表达水平,改善铜负荷引起的神经元损伤。

Objective: To explore the effect of Modified Gandou Decoction(MGDD) on the expression of ASM, Cer and p38 MAPK in the brain of TX mouse. Methods: One hundred and twenty TX mice were randomly divided into model group and MGDD group and selected 60 DL mice as the control group, MGDD group were given 24.5 g·kg-1·d-1 in gavage, Butyphthalide group were given 0.156 g·kg-1·d-1 in gavage. The control group and model group were given 100 m L·kg-1·d-1 in gavage, three months of age and five months of age were given two months and four months in gavage successionally, collect samples and then detect Acid sphingomyelinase(ASM), Ceramide(Cer), p38 mitogen activated protein kinase(p38 MAPK) of m RNA and proteins expression levels in TX mouse with MGDD treatment by RT-PCR, immunofluorescence and Western blot. Results: ASM, Cer, p38 MAPK of m RNA and proteins expression levels of model group in TX mouse was increased with the increasing of the months of age, and MGDD significantly decrease the expression of ASM, Cer, p38 MAPK in the brain of TX mouse(P<0.05, P<0.01) in dose-dependent and time-dependent manners. Conclusion: MGDD may improve neurotoxic induced by copper overload through reducing ASM, Cer, p38 MAPK of m RNA and proteins expression levels in TX mouse. MGDD relieve damage in brain tissue of TX mouse by regulating ceramide signaling pathway.