摘要
目的探讨健脾填精方对血管性痴呆(VD)小鼠脑组织关键蛋白的影响及作用机制。方法40只小鼠采用随机数字表法分为中药组15只,模型组15只,假手术组10只。模型组和中药组采用短暂双侧颈总动脉阻断法建立VD小鼠模型。中药组给予2.016 g/ml的健脾填精方溶液10 ml/(kg·d)灌胃,模型组和假手术组给予等量0.9%氯化钠溶液灌胃,3组均持续28 d。给药结束后对小鼠进行Morris水迷宫试验,利用定量蛋白质组学技术对小鼠脑组织蛋白进行质谱分析,筛选差异表达蛋白并进行生物信息学分析。结果水迷宫实验结果显示,试验第5天中药组逃避潜伏期时间[(38.12±2.15)s]显著短于模型组[(47.97±2.89)s],显著长于假手术组[(34.42±3.56)s],差异均有统计学意义(P<0.01);中药组跨越平台次数[(3.60±0.52)次]显著多于模型组[(2.40±0.52)次],显著少于假手术组[(4.70±0.82)次],差异均有统计学意义(均P<0.01);中药组停留在第Ⅲ象限时间[(16.71±1.19)s]显著长于模型组[(11.42±1.96)s],显著短于假手术组[(20.05±1.77)s],差异均有统计学意义(均P<0.01)。蛋白质组学共筛选出152个差异蛋白,模型组与中药组之间有65个差异蛋白;中药组小鼠脑组织中代谢性谷氨酸受体2(Grm2)、细胞色素c氧化酶亚基7C(Cox7c)、鸟苷酸环化酶可溶性亚基α-3(Gucy1a3)、蛋白偶联锌反转运蛋白Slc30a1(Slc30a1)等蛋白表达上调。结论健脾填精方可能通过影响小鼠脑组织中关键蛋白的表达来减轻兴奋性毒性、抗氧化应激、减轻线粒体功能障碍、抗动脉粥样硬化、调节细胞离子稳态等途径,从而改善VD。
Objective To investigate the effect and mechanism of Chinese medicine Jianpi Tianjing(JPTJ)formula on brain proteins in mice with vascular dementia(VD).Methods Forty ICR male mice were randomly divided into JPTJ group(n=15),model group(n=15)and sham group(n=10).The VD mouse model were induced by brief bilateral common carotid artery blockage in model group and the JPTJ group;while sham operation was performed in sham group.The mice in JPTJ group were gavaged with 10 ml JPTJ formula solution(2.016 g/ml),while the model group and sham group were given equal amount of saline.After the 28 d administration treatment,the mice were subjected to the Morris water maze test,then the brain tissues were harvested and the differentially expressed proteins were screened with mass spectrometry for quantitative proteomics and bioinformatics analysis.Results The water maze test showed that the latency time of escape was significantly shorter in the JPTJ group(38.12 s±2.15 s)than that in the model group(47.97 s±2.89 s),but longer than that in the sham-operated group(34.42 s±3.56 s)at D5(P<0.01).The number of platform crossings in the JPTJ group(3.60±0.52)was significantly more than that in the model group(2.40±0.52),but less than that in the sham group(4.70±0.82)(P<0.01).The time to stay in quadrant Ⅲ in the JPTJ group(16.71 s±1.19 s)was significantly higher than that in the model group(11.42 s±1.96 s),but significantly shorter than that in the sham group(20.05 s±1.77 s)(P<0.01).A total 152 differential proteins were screened out by proteomics,and there were 65 differential proteins between the model group and the JPTJ group.The expression of metabotropic glutamate receptor 2,cytochrome c oxidase subunit 7C,guanylate cyclase soluble subunit alpha-3,and proton-coupled zinc antiporter Slc30a1 were upregulated in the brain tissue of mice in JPTJ group.Conclusion Chinese medicine Jianpi Tianjing formula may improve VD in mice by affecting the expression of key proteins in brain tissue,which are associated with excitotoxicity,anti-oxidative stress,reducing mitochondrial dysfunction,anti-atherosclerosis,and regulating cellular ion homeostasis.
作者
龚娟芬
刘吉丹
房梦园
苏蒙
李伟光
何迎春
GONG Juanfen;LIU Jidan;FANG Mengyuan;SU Meng;LI Weiguang;HE Yingchun(Department of Geriatrics,Hangzhou Hospital of Traditional Chinese Medicine Affiliated to Zhejiang Chinese Medical University,Hangzhou 310007,China)
出处
《浙江医学》
CAS
2023年第9期968-974,I0005,I0006,共9页
Zhejiang Medical Journal
基金
杭州市科技发展计划项目(20201203B194)
浙江中医药科技计划项目(2022ZA104)。
关键词
健脾填精方
血管性痴呆
差异表达蛋白
蛋白质组学
中医药
Jianpi Tianjing formula
Vascular dementia
Differentially expressed protein
Proteomics
Traditional Chinese medicine
