在大鼠中15-LO/15-HETE参与缺氧对K_V1.5表达的抑制作用  

Hypoxia decreases K_V1.5 expression by 15-LO/15-HETE in rat

作　　者：褚晓杰[1] 张家宁[1] 郭蕾[1] 吴春铃[1] 张爽[1] 唐晓波[1] 朱大岭[1,2] 

机构地区：[1]哈尔滨医科大学药学院 [2]黑龙江省生物医药工程重点实验室-省部共建国家重点实验培育基地,黑龙江哈尔滨150086

出　　处：《中国药理学通报》2009年第3期326-331,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30370578;30470752;30870904);哈尔滨医科大学医学基础学科创新青年体基金

摘　　要：目的采用分子生物学技术从组织和细胞水平上观察阻断15-LO/15-HETE后,缺氧对KV1.5表达的影响。方法通过酶法分离、培养Wistar大鼠肺动脉血管平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)和大鼠肺动脉,应用Western blot和RT-PCR方法分别从蛋白质水平和mRNA水平上观察在15-LO阻断剂CDC和NDGA作用下,缺氧对KV1.5表达的影响。结果从组织和细胞水平上,用CDC和NDGA阻断15-LO即阻断了内源性15-HETE的产生,KV1.5的表达量在蛋白质水平和mRNA水平与未阻断组比较都增加。在阻断了内源性15-HETE的产生以后,加入外源性15-HETE,KV1.5的表达量减低。说明不仅内源性15-HETE参与诱导缺氧对KV1.5表达的影响,外源性15-HETE也同样能影响缺氧条件下KV1.5的表达量。但在阻断了内源性15-HETE的产生以后,加入外源性15-HETE,KV1.5的表达量减低。说明不仅内源性15-HETE参与诱导缺氧对KV1.5表达的影响,外源性15-HETE也同样能影响缺氧条件下KV1.5的表达量。结论上述结果表明,从大鼠肺动脉组织和细胞水平上,内源性15-HETE介导了缺氧对KV1.5表达的抑制作用。Aim To test the contribution of 15-HETE on expression of Kv1. 5 channel under hypoxia condition, using CDC or NDGA to block 15-LO/15-HETE, and to observe the effect of hypoxia on Kv1. 5 channel protein, mRNA expressions in cultured rat pulmonary arterial smooth muscle cells （PASMCs） and pulmonary arterials （ PAs ）. Methods Western blot, RT-PCR and 15-LO blockers, cinnamyl 3, 4-dihydroxy-[ alpha ] -cyanocinnamate （ CDC ） or nordihydroguiairetic acid （NDGA） were used to identify the role of endogenous 15-HETE on expression of Kv1. 5 channel in cultured rat pulmonary arterial smooth muscle cells （PASMCs） and PAs. Results （ 1 ） The expressions of Kv1. 5 channel protein and mRNA in PASMCs and PAs preteated with CDC or NDGA greatly increased than those of PASMCs under hypoxia group. （2） Exogenous 15-HETE added to PASMCs pretreated with CDC or NDGA greatly decreased the expression of Kv1. 5 than that of adding PASMCs pretreated with CDC or NDGA under hypoxia condition. Conclusion The down-regulation of Kv1. 5 channel expression caused by hypoxia is through endogenous 15-HETE.

关 键 词：15-HETE 缺氧 肺动脉平滑肌细胞 肺动脉 

分 类 号：R332[医药卫生—人体生理学] R322.121[医药卫生—基础医学]