热休克蛋白90在急性肺栓塞患者血浆刺激内皮细胞一氧化氮表达中的作用  

Effects of Hsp90 on the expression of eNOS in endothelial cells stimulated by pulmonary thromoembolism （PTE） patients＇ plasma

作　　者：张建初[1] 杨卫兵[1] 张晓菊[1] 夏蕾[1] 陶晓南[1] 白明[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院呼吸科,武汉430022

出　　处：《中华急诊医学杂志》2007年第2期185-188,共4页Chinese Journal of Emergency Medicine

基　　金：国家自然科学基金资助（50347009）;湖北省卫生厅自然科学基金资助项目（JX1B060）

摘　　要：目的探讨急性肺血栓栓塞（PTE）患者血浆刺激人脐静脉血管内皮细胞（HUVEC）内皮型一氧化氮合酶（eNOS）的表达过程中热休克蛋白90（Hsp90）的作用。方法分别用正常人血浆和PTE患者血浆孵育HUVEC，另设哥尔德霉素干预组，硝酸还原酶法测定各组细胞培养上清NO浓度，RT-PCR检测eNOS及Hsp90 mRNA的表达，放射免疫法测定cGMP的浓度。免疫共沉淀方法检测各组Hsp90和eNOS蛋白的共表达状况。结果PTE血浆处理组eNOS及Hsp90 mRNA表达水平较正常血浆对照组增强；NO浓度亦明显高于正常对照组及哥尔德霉素处理组[（16．62±3．14）μmol／L vs（5．16±1．98）μmol/L，（9．34±1．09）μmol/L]，差异均具有统计学意义（t=13．2,P〈0．01；t=15．6，P〈0．01）；PTE血浆处理组cGMP浓度亦明显高于正常对照组及哥尔德霉素处理组[（3．76±1．04）pmo;/L体（2．01±0．98）pmol/L，（1．87±0．99）pmo;/L]，差异均具有统计学意义（t=11．6，P〈0．01；t=12．1，P〈0．01）；免疫共沉淀检测结果显示Hsp90和eNOS蛋白共表达。结论 PTE患者血浆可以刺激血管内皮细胞Hsp90 mRNA及eNOS表达上调，抑制Hsp90可下调eNOS生物活性从而减少NO的产生。Objective To observe the effects of heart shoch protein （Hsp90） on the expression of eNOS in endothelial cells stimulated by PET patients＇ plasma. Methods Human umbilical vein endothelial cells were cultured with plasma of PTE patients （experiment group） and with plasma of normal people （control group）. The NO and cyclic guansine monophosphate （cGMP） concentration in cell cultured supematant were detected with nitrate reduetase and radio-immunity, respectively. The expression of Hsp90 and eNOS mRNA were detected by RT-PCR. The NO and eGMP production were measured after using Geldanamyein to block the action of Hsp90, and the protein expression of Hsp90 and eNOS in experimental group and control group were detected with coimmunoprecipitatian. Results The expression of Hsp90 and eNOS mRNA in experimental group were higher than that in control group. The NO concentration in experimental group was （16.62 ± 3.14） μmol/L, whereas it was （5.16 ± 1.98） μmol/L in control group; it and （9.34 ± 1.09） μmol/L in the C, eldanamyein treated group, and compared with the experimental group, there existed difference （ t = 13.2, P 〈0.01; t = 15.6, P 〈0.01） . The eGMP concentration in experimental group were higher than that in control group and Geldanamyein treated group C （3.76 ± 1.04） pmol/L vs. （2.01± 0.98） pmol/L, （ 1.87± 0.99） pmol/L], and there was significant difference （ t = 11.6, P 〈 0.01; t = 12.1, P 〈 0.01） . At the same time, the result of co- showed there was cecxpression of Hsp90 and eNOS proteins. Conclusions FTE patients＇ plasma could up-regulate the expression of eNOS and HspgO in endothelial cell, and inhibiting the role of Hsp90 resulted in the down-regulated expression of eNOS, so reduced production of NO.

关 键 词：内皮型一氧化氮合酶 急性肺血栓栓塞 血浆 热休克蛋白90 

分 类 号：R686[医药卫生—骨科学]