基于蛋白质组学探讨泻南补北法对去卵巢雌鼠脑组织蛋白表达水平的影响  

Effects of reducing the south and reinforcing the north method on brain protection in ovariectomized female rats based on proteomics

作　　者：吴宏进[1] 戴薇薇[1] 王利波[1] 张婕 王成龙[1] WU Hong-jin;DAI Wei-wei;WANG Li-bo;ZHANG Jie;WANG Cheng-long(Central Laboratory for Science and Technology,Longhua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院科技中心实验室,上海200032

出　　处：《中华中医药杂志》2022年第7期4026-4032,共7页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：上海市自然科学基金项目(No.16ZR1447300);上海中医药大学第十九期校级课程建设项目(No.2019SHUTCM109)。

摘　　要：目的:基于蛋白质组学方法研究泻南补北法对去卵巢雌鼠脑组织蛋白表达水平的影响,进而探索其脑保护机制。方法:将40只雌性SD大鼠随机分为4组,对照组(等容积蒸馏水)、模型组(等容积蒸馏水)、中药组(中药复方,5.46g·kg^(-1)·d^(-1))、西药组(戊酸雌二醇,0.1mg·kg^(-1)·d^(-1)),每组10只。给药6周后,分析各组大鼠体质量、性激素水平。取大鼠脑组织用非标记蛋白质组学联合液性色谱质谱(LC-MC/MC)进行蛋白质组学分析,并将筛选出差异蛋白进行Western blot法进行检测验证。结果:共得到4678个蛋白质。对照组和模型组共有87个差异蛋白(上调37个,下调50个);中药组和模型组共有242个差异蛋白(上调95个,下调147个)。中药组或西药组与对照组变化趋势相一致的差异蛋白质有22个,这些差异蛋白质主要涉及脂肪酸代谢、生物氧化、信号转导、胆固醇代谢、免疫反应、应激反应及细胞凋亡等生物学过程;Western blot方法检测相关差异蛋白结果,泻南补北法对去卵巢大鼠脑组织中的GAT1、ApoE、JNK和p-JNK蛋白具有一定的调节作用(P<0.05),与蛋白质组学分析结果一致。结论:泻南补北法可能通过调节GAT1、ApoE、JNK、p-JNK等蛋白水平,影响脂质代谢、免疫反应、应激反应等,从而实现对去卵巢大鼠的脑部保护作用。Objective:To explore the effects of reducing the south and reinforcing the north(RSRN)method on differential brain tissue proteins in ovariectomized(OVX)rats,further to analyze its brain protection mechanism.Methods:Forty eight-week-old female Sprague-Dawley rats were randomly divided into four groups:SHAM group(sham-operation,equal volume ddH_(2)O);OVX group(ovariectomized,equal volume ddH_(2)O);TCM group(ovariectomized,RSRN:5.46 g·kg^(-1)·d^(-1));and EV group(ovariectomized,estradiol valerate:0.1 mg·kg^(-1)·d^(-1)).After administration for 6 weeks,we measured body weight,serum FSH,and LH.Rat brain tissues were analyzed using LC-MS/MS.Subsequently,we performed bioinformatics analysis and screened differential proteins for detection and verification through Western blot analysis.Results:A total of 4678 proteins were identified and quantified.There were 87 differential proteins in the SHAM and OVX group;among them,37 were up-regulated and 50 were down-regulated.There were 242 differential proteins in the TCM and OVX groups;among them,95 were up-regulated and 147 were down-regulated.Twenty-two proteins in the TCM group or EV group that showed a similar trend to the OVX group,which were mainly involved in lipid metabolism,biological oxidation,signal transduction,cholesterol metabolism,immune response,stress response,and cell apoptosis.Western blot method was used to detect the results of related differential proteins.The method of RSRN had an regulation effect on GAT1,ApoE,JNK and p-JNK proteins in the brain tissue of ovariectomized rats(P<0.05),which was consistent with the results of proteomic analysis.Conclusion:RSRN may protect the brains of OVX hyperlipemia rats by exerting an effect on lipid metabolism,immune response,stress response through regulating GAT1,ApoE,JNK,and p-JNK proteins.

关 键 词：泻南补北 围绝经期 脑保护 蛋白质组学 中医药 神经系统疾病 

分 类 号：R285.5[医药卫生—中药学]