高脂血症早期血管脂联素抵抗受体前机制的探讨  

Mechanism of vascular adiponectin resistance in early phase of hyperlipidemia at the pre-receptor Level

作　　者：樊森 戴建立 王晓明[1] 李榕[1] 

机构地区：[1]第四军医大学西京医院老年病科,陕西西安710032

出　　处：《心脏杂志》2017年第6期631-635,共5页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(81270401;81470413);陕西省自然科学基础研究计划项目资助(2014JZ010);陕西省卫计委科研项目资助(2016D002)

摘　　要：目的明确高脂血症早期血管脂联素(APN)抵抗是否与高脂血浆中血管活性物质与APN结合并抑制其生物活性有关。方法将20只雄性SD大鼠随机分为对照组和高脂血症组,每组10只,分别饲以正常及高脂饮食8周。喂养结束后,麻醉采血检测血浆中总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)和APN的水平。采用免疫共沉淀法沉淀血浆中的APN,质谱分析法分析血浆中能够与APN结合的血管活性物质。将初步得到的血管活性物质与重组人APN(r APN)共孵育,观察孵育后的r APN对人脐静脉内皮细胞(HUVECs)中腺苷酸活化的蛋白激酶(AMPK)磷酸化(p-AMPK)水平的影响。结果与喂养正常饲料组相比,以高脂饲料喂养8周后,大鼠TC、TG、HDL-C、LDL-C、APN及血糖的含量显著升高(P<0.05或P<0.01)。将正常及高脂饲料喂养大鼠的血浆用APN抗体进行免疫共沉淀并进一步行质谱分析后,发现载脂蛋白(Apo)A1、ApoC1、对氧磷酶(PON)1等18种血管活性物质与高脂血浆中APN结合显著增多(P<0.01)。其中ApoA1在血浆中含量显著升高(P<0.05),此外,ApoA1及ApoC1可显著抑制r APN对HUVECs中7-AMPK水平(P<0.01或P<0.05)。结论高脂血症早期大鼠血管APN抵抗与高脂血浆中Apo水平升高、结合并抑制APN活性有关。AIM To identify whether vascular activity factors that can combine and inhibit the ability of APN are relate to vascular APN resistance in early phase hyperlipidemia. METHODS Twenty male Sprague Dawley rats were divided equally into control group and hyperlipidemia group randomly, receive regular diet or high-fat diet for 8 weeks, then triglyceride, total cholesterol, high-density lipoprotein, low-density lipoprotein in circulation were determined. Plasma from regular diet and high-fat diet rats was applied co-immunoprecipitation with anti-APN antibody to precipitate plasma APN then followed mass spectrometric analysis to determine which vascular activity factors can combine with APN. Pre-incubated rAPN with vascular activity factors which discovered in former step and observed the AMPK activation effect of rAPN in HUVECs. RESULTS Compared with rats fed with regular diet rats fed with high-fat diet triglyceride, total cholesterol, high-density lipoprotein, low-density lipoprotein and APN levels in circulation were obviously raised （P 〈 0. 05 or P 〈 0. 01 ） after 8 weeks. Discovered ApoA1, ApoC1, PON1 along with other fifteen vascular activity factors can combine APN and the ability of combinationsignificantly increased in hyperlipidemia plasma （P 〈 0. 01 ）. Among the eighteen vascular activity factors ApoA1 significantly increased in hyperlipidemia plasma （P 〈 0. 05 ）, ApoA1 and ApoC1 can significantly inhibit the ability of rAPN to phosphorylate AMPK in HUVECs （ P 〈 0.01 or P 〈 0.05 ）. CONCLUSION Increased apolipoprotein that can combine and inhibit the activity of APN in hyperlipidemic plasma is likely responsible for vascular APN resistance in early phase hyperlipidemia.

关 键 词：脂联素抵抗 高脂血症 载脂蛋白A1 载脂蛋白C1 

分 类 号：R587.1[医药卫生—内分泌]