安徽医科大学学报
安徽醫科大學學報
안휘의과대학학보
ACTA UNIVERSITY MEDICINALIS ANHUI
2015年
5期
565-568
,共4页
张洁%沈兵%桑大成%杜鹃%丁圣刚
張潔%瀋兵%桑大成%杜鵑%丁聖剛
장길%침병%상대성%두견%정골강
钙激活钾通道%瞬时受体电位离子通道 4%14,15-EET%气管平滑肌
鈣激活鉀通道%瞬時受體電位離子通道 4%14,15-EET%氣管平滑肌
개격활갑통도%순시수체전위리자통도 4%14,15-EET%기관평활기
Ca2 + -activated K + channel%transient receptor potential vanilloid 4%14,15-EET%airway smooth mus-cle
目的:探讨花生四烯酸细胞色素 P450(CYP)表氧化酶代谢产物14,15-环氧化二十碳三烯酸(14,15-EET)对小鼠气管平滑肌收缩功能的影响及其机制。方法采用离体气管实验,通过运用特异性钙激活钾通道阻断剂和瞬时受体电位离子通道4(TRPV4)通道阻断剂,观察14,15-EET 对卡巴胆碱引起的小鼠气管收缩的影响;采用免疫共沉淀实验检测 TRPV4通道蛋白与小电导钙激活钾通道(SKCa )蛋白在小鼠气管平滑肌组织中的相互作用。结果与对照组相比,300 nmol/ L 14,15-EET 预处理小鼠气管后,卡巴胆碱引起的收缩显著减弱;大、中电导钙激活钾通道阻断剂 IbTX 和TRAM34没有显著影响14,15-EET 对卡巴胆碱引起的小鼠气管收缩的抑制效应;而 SKCa阻断剂 Apamin 和 TRPV4通道阻断剂 RN-1734都分别显著阻断了14,15-EET 对卡巴胆碱引起的小鼠气管收缩的抑制作用。免疫共沉淀结果显示TRPV4通道蛋白和 SKCa通道蛋白可以彼此相互共沉淀。结论在小鼠气管平滑肌中,14,15-EET 通过 TPRV4-SKCa钙信号复合物调节气管平滑肌的收缩。
目的:探討花生四烯痠細胞色素 P450(CYP)錶氧化酶代謝產物14,15-環氧化二十碳三烯痠(14,15-EET)對小鼠氣管平滑肌收縮功能的影響及其機製。方法採用離體氣管實驗,通過運用特異性鈣激活鉀通道阻斷劑和瞬時受體電位離子通道4(TRPV4)通道阻斷劑,觀察14,15-EET 對卡巴膽堿引起的小鼠氣管收縮的影響;採用免疫共沉澱實驗檢測 TRPV4通道蛋白與小電導鈣激活鉀通道(SKCa )蛋白在小鼠氣管平滑肌組織中的相互作用。結果與對照組相比,300 nmol/ L 14,15-EET 預處理小鼠氣管後,卡巴膽堿引起的收縮顯著減弱;大、中電導鈣激活鉀通道阻斷劑 IbTX 和TRAM34沒有顯著影響14,15-EET 對卡巴膽堿引起的小鼠氣管收縮的抑製效應;而 SKCa阻斷劑 Apamin 和 TRPV4通道阻斷劑 RN-1734都分彆顯著阻斷瞭14,15-EET 對卡巴膽堿引起的小鼠氣管收縮的抑製作用。免疫共沉澱結果顯示TRPV4通道蛋白和 SKCa通道蛋白可以彼此相互共沉澱。結論在小鼠氣管平滑肌中,14,15-EET 通過 TPRV4-SKCa鈣信號複閤物調節氣管平滑肌的收縮。
목적:탐토화생사희산세포색소 P450(CYP)표양화매대사산물14,15-배양화이십탄삼희산(14,15-EET)대소서기관평활기수축공능적영향급기궤제。방법채용리체기관실험,통과운용특이성개격활갑통도조단제화순시수체전위리자통도4(TRPV4)통도조단제,관찰14,15-EET 대잡파담감인기적소서기관수축적영향;채용면역공침정실험검측 TRPV4통도단백여소전도개격활갑통도(SKCa )단백재소서기관평활기조직중적상호작용。결과여대조조상비,300 nmol/ L 14,15-EET 예처리소서기관후,잡파담감인기적수축현저감약;대、중전도개격활갑통도조단제 IbTX 화TRAM34몰유현저영향14,15-EET 대잡파담감인기적소서기관수축적억제효응;이 SKCa조단제 Apamin 화 TRPV4통도조단제 RN-1734도분별현저조단료14,15-EET 대잡파담감인기적소서기관수축적억제작용。면역공침정결과현시TRPV4통도단백화 SKCa통도단백가이피차상호공침정。결론재소서기관평활기중,14,15-EET 통과 TPRV4-SKCa개신호복합물조절기관평활기적수축。
Objective To provide a mechanistic insight into how 14,15-epoxyeicosatrienoic acid (14,15-EET) which is a product of cytochrome P450 epoxygenase regulates mouse airway smooth muscle contraction. Methods Isolated mouse tracheal tension was measured in vitro. Mouse tracheal rings were treated by Ca2 + -activated K +channel blockers or transient receptor potential vanilloid 4 (TRPV4) channel blocker. The changes of relaxation caused by 14,15-EET in tracheal rings were recorded after the tracheal rings were contracted by carbachol in con-centration-dependent fashion. Co-immunoprecipitation was used to examine the physical interaction between TRPV4 and SKCa in airway smooth muscle. Results Tracheal tension measurement showed that compared with the control group, carbachol-induced contraction in 300 nmol/ L 14,15-EET pretreatment group was significantly reduced. BK-Ca and IKCa blockers did not affect the effect of 14,15-EET on carbachol-induced tracheal contraction. However, SKCa and TRPV4 blockers notably inhibited the effect of 14,15-EET on carbachol-induced tracheal contraction, re-spectively. Conclusion 14,15-EET regulates airway smooth muscle contraction via TRPV4-SKCa signal complex.