HSP70及其磷酸化在下肢动脉硬化闭塞症中的表达及临床意义

霍鑫 曹治婷 蔡许超

[摘要] 目的 分析下肢动脉硬化闭塞症（LEASO）患者不同阶段热休克蛋白70（HSP70）及其磷酸化水平的差异及其临床意义。 方法 收集2017年9月至2019年12月在柳州市人民医院收治的80例下肢动脉硬化闭塞症（LEASO）患者的血样，按踝肱指数（ABI）分为A组（ABI≤0.430）和B组（ABI>0.430）。采用酶联免疫吸附法（ELISA）测定本院患者各阶段HSP70的表达，分析两组HSP70浓度及ABI的差异。同时采集4例正常股动脉标本（正常组）和4例动脉粥样硬化性股动脉标本（病变组），用免疫印迹法检测HSP70及其磷酸化HSP70（Ser15）、磷酸化HSP70（Ser78）和磷酸化HSP70（Ser82）。对正常组与病变组的蛋白表达水平进行统计学分析。 结果 A组HSP70浓度为（46.731±12.986）ng/mL，ABI为（0.231±0.056）。B组HSP70浓度为（67.104±18.262）ng/mL，ABI为（0.622±0.065）。正常组HSP70蛋白表达及磷酸化水平分别为（0.812±0.131） ng/mL、（0.643±0.107） ng/mL、（0.918±0.206） ng/mL和（0.891±0.168） ng/mL;病變组蛋白表达分别为（0.463±0.072） ng/mL、（0.424±0.081） ng/mL、（0.393±0.065） ng/mL和（0.572±0.101） ng/mL，组间比较差异有统计学意义（P<0.05）。 结论 血清HSP70浓度与LEASO严重程度呈负相关，而HSP70浓度与ABI值呈正相关。病变组HSP70含量及其磷酸化水平明显低于正常组，可能与动脉粥样硬化的发生发展密切相关。

[关键词] 下肢动脉硬化闭塞症;热休克蛋白70;磷酸化;踝肱指数

[中图分类号] R543.5          [文献标识码] B          [文章编号] 1673-9701（2021）07-0062-04

Expression and clinical significance of HSP70 and its phosphorylation in arteriosclerosis obliterans of lower extremities

HUO Xin1   CAO Zhiting2   CAI Xuchao3

1.Department of Vascular Surgery，Liuzhou People′s Hospital in Guangxi Zhuang Autonomous Region， Liuzhou   545005， China; 2.Department of Radiology，Liuzhou People′s Hospital in Guangxi Zhuang Autonomous Region， Liuzhou   545005，China; 3.Department of Vascular Surgery， Hangzhou Geriatric Hospital， Hangzhou   310022， China

[Abstract] Objective To analyze the differences in heat shock protein 70（HSP70）and its phosphorylation levels in patients with lower extremity arteriosclerosis obliterans（LEASO） at different stages， and its clinical significance. Methods Blood samples of 80 patients with lower extremity arteriosclerosis obliterans（LEASO） who were admitted to Liuzhou People′s Hospital from September 2017 to December 2019 were collected and divided into group A （ABI≤0.430） and group B （ABI>0.430） according to the ankle-brachial index （ABI）. Enzyme-linked immunosorbent assay （ELISA） was used to determine the expression of HSP70 in patients in our hospital at various stages. The differences in HSP70 concentration and ABI between the two groups were analyzed. At the same time， four normal femoral artery specimens（normal group） and four atherosclerotic femoral artery specimens（pathological group） were collected. The HSP70 and its phosphorylated HSP70（Ser15）， phosphorylated HSP70（Ser78） and phosphorylated HSP70 （Ser82） were detected by Western blotting. The protein levels of the normal group and the diseased group were statistically analyzed. Results The concentration of HSP70 in group A was（46.731±12.986）ng/mL， and the ABI was （0.231±0.056）. The concentration of HSP70 in group B was（67.104±18.262）ng/mL， and the ABI was （0.622±0.065）. The protein expression and phosphorylation level of HSP70 in the normal group were （0.812±0.131）ng/mL， （0.643±0.107）ng/mL， （0.918±0.206）ng/mL and （0.891±0.168） ng/mL， respectively. The protein expression and phosphorylation level of HSP70 in the pathological group was（0.463±0.072）ng/mL， （0.424±0.081）ng/mL， （0.393±0.065）ng/mL and（0.572±0.101）ng/mL， there was significant difference between the two groups（P<0.05）. Conclusion Serum HSP70 concentration is negatively correlated with LEASO severity， while HSP70 concentration is positively correlated with ABI value. The HSP70 content and phosphorylation level of the diseased group is significantly lower than that of the normal group，which may be closely related to the occurrence and development of atherosclerosis.

[Key words] Lower extremity arteriosclerosis obliterans; Heat shock protein 70; Phosphorylation; Ankle-brachial index

动脉粥样硬化是导致心、脑、四肢缺血的主要心血管疾病，严重者可导致梗死。动脉粥样硬化已成为威胁人类健康的第一大杀手和世界上人类死亡的主要原因[1]。下肢动脉硬化闭塞症（Lower extremity arteriosclerosis obliterans，LEASO）是一种慢性进行性疾病，其表现为动脉粥样硬化，使下肢动脉内膜增厚，管腔变窄甚至闭塞，进而导致患肢间歇性跛行、静息痛甚至溃疡等一系列症状和体征坏死。动脉粥样硬化，尤其是LEASO，是一个复杂的炎症过程，是冠狀动脉疾病的主要潜在原因。目前对下肢动脉粥样硬化闭塞症发病机制的研究表明，内皮功能障碍是动脉粥样硬化的早期事件，与动脉粥样硬化斑块的发展密切相关[2]。