基于蛋白质组学探讨痰湿壅盛高血压大鼠下丘脑对高脂饮食应答的改变

Role of Hypothalamic Response to High Fat Diet in Hypertensive Rats with Excessive Accumulation of Phlegm-Dampness Based on Proteomics

文章基于蛋白质组学方法旨在探讨痰湿壅盛高血压病理状态下下丘脑对高脂饮食应答的改变。自发性高血压大鼠(SHR) 10只、血压正常的WKY大鼠10只,给以高脂饲料喂养24周。取下丘脑进行双向电泳、硝酸银染色、筛选差异蛋白质点提交质谱鉴定,采用MALDI-TOF-TOF质谱仪进行质谱分析、肽指纹图谱做蛋白质鉴定,并以western blot验证结果。结果显示,高脂饮食24周后,大鼠体重、血压、胆固醇、胰岛素、血糖/胰岛素比均有不同程度的改变。SHR、WKY大鼠共分离出3280&3844个蛋白点,其中SHR与WKY大鼠比较,105个蛋白质表达降低,54个蛋白质表达增加。差异蛋白分别与细胞骨架、神经元结构功能、能量代谢、应激、肿瘤相关蛋白等有关。

Objective:To explore the molecular basis of responsiveness of hypertensive hypothalamus with excessive accumulation of phlegm-dampness to high fat diet based on proteomics.Methods:Ten spontaneously hypertensive rats(SHR)and 10 normotensive WKY rats were fed with high fat diet for 24 weeks.The hypothalami were removed and the differentially expressed profile in hypothalami was identified using two-dimension electrophoresis coupled with MALDI-TOF-TOF mass spectrometry technique.The results were verified by Western blot.Results:After chronic high fat diet,the weight,blood pressure,cholesterol,insulin,ratio of blood glucose/insulin were changed in varying degrees in SHR and WKY rats.A total of 3280&3844 protein spots were isolated from SHR and WKY rats.Among them,105 proteins were decreased and 54 proteins were increased in SHR compared with WKY rats.The differentially expressed proteins were enriched in cytoskeleton,neuronal structure and function,energy metabolism,stress and tumor-related protein.Conclusion: