脾阴虚证脾失健运大鼠回肠组织中相关蛋白表达差异及其调控机制的实验研究

发布时间：2018-12-14 23:45

【摘要】：目的:探讨脾阴虚证脾失健运状态下,大鼠回肠组织中相关差异蛋白的表达及其调控机制。方法:根据分层随机原则,将36只SPF级健康Sprague-Dawley(SD)大鼠,随机分成健康对照组、脾阴虚证模型组、中药反证组共3组,每组各12只。采用经典复合因素法塑造脾阴虚证脾失健运大鼠模型。行逆转录聚合酶链反应(RT-PCR)法及蛋白免疫印迹(Western blot)法,分别对课题组前期蛋白组学鉴定出的过氧化氢酶(Catalase,CAT)、顺乌头酸水合酶(Aconitate hydratase,ACO2)、钙联结蛋白(Calnexin,Canx)、Papss 2(Papss 2 69KD protein)、热休克蛋白90(heat shock protein 90KD,HSP 90)5个特异性表达蛋白进行检测。结果:(1)半定量分析比较各组大鼠回肠组织差异表达蛋白mRNA表达水平发现,与健康对照组比较,脾阴虚模型组回肠组织中CAT、Canx、ACO2及Papss 2蛋白的mRNA表达均显著上升(P0.05),而HSP 90蛋白的mRNA表达显著下调(P0.05),中药反证组各指标均未出现明显差异(P0.05);(2)半定量分析比较各组大鼠回肠组织差异表达蛋白表达水平发现,与健康对照组比较,脾阴虚模型组回肠组织中CAT、Canx、ACO2及Papss 2蛋白含量均显著上升(P0.05),其中以CAT上调的最为显著(P0.01),而HSP 90蛋白含量显著下调(P0.05),中药反证组各指标均未出现明显差异(P0.05)。结论:脾阴虚脾失健运状态下,大鼠回肠组织中可能存在以线粒体损伤为基础的细胞膜过氧化损伤、Ca2+稳态失衡、类乙醇毒害、过度硫酸化、神经元损伤及能量和遗传物质变化等一系列病理改变。
[Abstract]:Objective: to investigate the expression and regulation mechanism of differentially expressed proteins in ileum of rats with spleen deficiency syndrome. Methods: according to the principle of stratification and randomization, 36 healthy Sprague-Dawley (SD) rats of SPF grade were randomly divided into three groups: the healthy control group, the model group of spleen yin deficiency syndrome, and the Chinese medicine reverse syndrome group, each with 12 rats in each group. The rat model of spleen deficiency was established by classical compound factor method. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot (Western blot) were used to detect catalase (Catalase,CAT), cisaconitate hydratase (Aconitate hydratase,ACO2), which were identified by proteomics. The expression of Calnexin,Canx), Papss 2 (Papss 2 69KD protein), heat shock protein 90 (heat shock protein 90 KDX HSP90) was detected. Results: (1) the expression level of differentially expressed protein mRNA in ileum tissue of rats in each group was compared by semi-quantitative analysis. Compared with healthy control group, CAT,Canx, in ileum of spleen yin deficiency model group was found. The mRNA expression of ACO2 and Papss 2 protein increased significantly (P0.05), while the mRNA expression of HSP 90 protein decreased significantly (P0.05). (2) the expression level of differentially expressed protein in ileum of rats in each group was compared by semi-quantitative analysis. Compared with the healthy control group, the contents of CAT,Canx,ACO2 and Papss 2 protein in ileum of spleen yin deficiency model group were significantly increased (P0.05). Among them, CAT up-regulated the most significantly (P0.01), while the content of HSP 90 protein decreased significantly (P0.05). Conclusion: under the condition of spleen yin deficiency and spleen incapacitation, there may be membrane peroxidation damage, Ca2 homeostasis, ethanol-like toxicity and excessive sulfation in ileum tissue of rats based on mitochondrial damage. A series of pathological changes, such as neuronal damage and changes in energy and genetic material.
【作者单位】：辽宁中医药大学;
【基金】：国家自然科学基金面上项目(81373502) 辽宁省高等学校优秀人才支持计划项目(LR2013043)
【分类号】：R228

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