针刺“太溪”穴对肾脏组织双向电泳图谱影响的实验研究

目的:利用双向电泳技术,观察针刺足少阴肾经原穴"太溪"穴前后肾脏组织双向电泳图谱的变化。方法:健康8周龄雄性Wistar大鼠18只,随机分为空白组、非经非穴组和针刺组,每组6只。非经非穴组每日针刺非穴点,针刺组每日电针双侧"太溪"穴,空白组大鼠在相同时间只予固定,不予针刺。1周后取出各组大鼠肾脏组织,提取肾脏组织总蛋白,进行双向电泳,比较分析各组肾脏组织双向电泳图谱的变化。结果:空白组和非经非穴组肾脏蛋白质未发现差异蛋白点;针刺组较空白组肾脏蛋白质显示9个3倍以上上调差异蛋白点,未发现下调蛋白质。结论:初步建立了大鼠肾脏组织蛋白质双向电泳的技术方法;针刺组大鼠蛋白质与空白组、非经非穴组存在明显差异,差异点的发现为下一步进行质谱分析奠定了基础。

Research on the Effect of Two Dimensional Electrophoresis of Kidney After Acupuncturing Yuan-acupoint "Taixi"

Objective:To research the changes of between variation of kidney proteins and acupuncture after acupuncturing yuan-acupoint(Taixi)of the Kidney Meridian of Foot-shaoyin before and after treatment using two dimensional electrophoresis.Methods:18 adult male Wistar rats were randomly divided into normal control group(n=6),sham acpoints group(n=6)and acupuncture group(n=6).Rats of acupuncture group were electro-acupunctured both side of Taixi(2Hz) for 20 min once a day.Rats of sham acpoints group were electro-acupunctured both side of sham non-points(2Hz) for 20 min once a day.The rats of normal control group took only the experiments in bag,but no acupuncture.After 7 days,the kidneys of rats were removed and the total protein of kidney was isolated.After protein sample preparation,Two-dimensional gel electrophoresis(2-DE) was performed and the changes of two-dimensional gel electrophoresis every group were compared with each other using Image MasterTM 2D Platinum.Results:No abnormal proteins was observed within normal control group and sham acpoints group.After acupuncture,9 proteins showed 3-fold increase in kidney of acupuncture group.Conclusion:The two-dimensional electrophoresis of kidney method for detection proteins was preliminarily developed.There are clear differences between acupuncture group and the other groups.The foundation of differentially expressed proteins provides a solid basis for the futher study by Liquid Chromatography-Mass Spectrometry.