肝豆汤改良方对TX小鼠脑组织XIAP、Caspase-9和Caspase-3蛋白及其mRNA表达的影响  

Effects of Modified Gandou Decoction on Protein and mRNA Expression of X-linked Inhibitor of Apoptosis,Caspase-9,and Caspase-3 in Brain Tissue of TX Mice

作　　者：赵雯[1,2] 马艳红[1,2] 韩咏竹[2] 程楠[2] 饶娆[2] 王训[2] 

机构地区：[1]安徽中医药大学,安徽合肥230038 [2]安徽中医药大学神经病学研究所附属医院,安徽合肥230061

出　　处：《安徽中医药大学学报》2015年第4期58-64,共7页Journal of Anhui University of Chinese Medicine

基　　金：国家自然科学基金项目(81173212)

摘　　要：目的探究肝豆汤改良方抑制肝豆状核变性(Wilsons disease,WD)脑神经细胞凋亡的分子机制。方法选取1月龄TX小鼠120只,随机分为TX模型组(40只)、肝豆汤改良方组(40只)、丁苯酞组(20只),另选DL小鼠40只作为正常组;每组再分成2组,各20只,分别予以生理盐水、肝豆汤改良方、丁苯酞灌胃2个月和4个月。分别采用免疫组织化学法和Western blot法检测小鼠脑组织中X-连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis,XIAP)、胱冬肽酶-9(Caspase-9)及胱冬肽酶-3(Caspase-3)蛋白表达水平,采用RT-PCR技术检测小鼠脑组织中XIAP、Caspase-9及Caspase-3mRNA表达水平。结果免疫组织化学检测显示,肝豆汤改良方组XIAP表达水平较同月龄模型组明显增多,Caspase-9和Caspase-3表达水平较同月龄模型组下降,5月龄和3月龄肝豆汤改良方组XIAP和Caspase-3表达水平有所差异。Western blot检测结果显示,月龄因素对脑组织XIAP、Caspase-9、Caspase-3蛋白表达水平的主效应均无统计学意义(P>0.05);月龄因素和分组因素对3种蛋白表达水平的交互作用均无统计学意义(P>0.05);对于同月龄小鼠,模型组脑组织XIAP表达水平显著低于正常组(P<0.05),Caspase-9和Caspase-3蛋白表达水平显著高于正常组(P<0.05);肝豆汤改良方组XIAP表达水平显著高于模型组(P<0.05),Caspase-9和Caspase-3蛋白表达水平显著低于模型组(P<0.05)。RT-PCR检测结果显示,月龄因素对小鼠脑组织XIAP mRNA表达水平的主效应无统计学意义(P>0.05),但对Caspase-9、Caspase-3mRNA表达水平的主效应具有统计学意义(P<0.05);月龄因素和分组因素对XIAP、Caspase-9、Caspase-3mRNA表达水平的交互作用均无统计学意义(P>0.05)。对于相同月龄小鼠,模型组脑组织XIAP mRNA表达水平显著低于正常组(P<0.05),而Caspase-9、Caspase-3mRNA表达水平显著高于正常组(P<0.05);与模型组比较,丁苯酞组和肝豆汤改良方组XIAP mRNA表达水平显著升高(P<0.05)Objective To investigate the molecular mechanism of action of modified Gandou Decoction （MGD） in inhibiting cerebral neuronal apoptosis in Wilson＇s disease. Methods hundred and twenty 1- month-old TX mice were randomly and equally divided into model group, MGD group, and butylphthalide group. Mice in each group were equally divided into two subgroups, which were given saline solution, MGD, or butylphthalide by gavage for 2 and 4 months, respectively. Forty DL mice were enrolled into normal group. The protein expression of X-linked inhibitor of apoptosis （XIAP）, Caspase-3, and Caspase- 9 in the brain tissue of mice was measured using immunohistochemical assay and Western blot; the mRNA expression of XIAP, Caspase-3, and Caspase-9 in the brain tissue of mice was determined using RT-PCR. Results According to the immunohistochemical assay, the MGD-treated mice had significantly higher ex- pression of XIAP protein but significantly lower expression of Caspase-9 and -3 protein than the model mice of the same age. Moreover, there were differences in the protein expression of XIAP and Caspase-3 be- tween the 5-month-old and 3-month-old mice in MGD group. According to the results of Western blot, there was no significant relationship of age with the protein expression of XIAP, Caspase-9, and Caspase- 3 in brain tissue （P〉0. 05）; there was also no significant interaction between age and group for the ex- pression of these three proteins （P〉0.05）; the model mice had significantly lower expression of XlAP protein but significantly higher expression of Caspase-9 and -3 protein than the normal mice of the same age （all P〈0.05） ; the MGD-treated mice had significantly higher expression of XIAP protein but significantly lower expression of Caspase-9 and -3 protein than the model mice of the same age （all P〈0.05）. Accord- ing to the results of RT-PCR, there was significant relationship of age with the mRNA expression of Caspase-9 and -3 （both P〈0.05） rather than XIAP （P〉0.05） ; there

关 键 词：肝豆状核变性 肝豆汤改良方 TX小鼠 神经细胞损伤 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学] R742.4[医药卫生—中医学]