蛋白激酶cδ可能参与肥大心肌细胞转向凋亡

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1、生理学报ActaPhysiologicaSinica,June25,2006,58(3):269-274269http://www.actaps.com.cn研究论文蛋白激酶Cδ可能参与肥大心肌细胞转向凋亡郭万刚，余志斌*，谢满江第四军医大学航空航天生理学教研室，西安710032摘要：为了探讨肥大心肌细胞对凋亡刺激的易感性及蛋白激酶Cδ(proteinkinaseCδ,PKCδ)在其中的作用，以内皮素-1(endothelin-1,ET-1)处理原代培养的新生大鼠心肌细胞，诱导心肌细胞肥大；再用血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)作为细胞凋亡诱导因子，采用鬼笔环肽(phallo

2、idin)荧光染色与细胞面积测量两种方法检测心肌肥大，Hoechst33258荧光染色检测细胞凋亡。结果显示：(1)1与10nmol/LET-1作用48h，心肌细胞肌原纤维排列整齐、染色增浓，随ET-1浓度增加而愈加明显，心肌细胞表面积分别增加42.5%和67.3%，以此作为轻度和中度心肌细胞肥大模型。(2)正常、轻度肥大与中度肥大心肌细胞受1nmol/LAngⅡ处理24h后,凋亡率分别为(15.54±1.32)%、(20.65±1.40)%与(29.33±3.52)%，三组之间有显著差异(P<0.05)。(3)受AngⅡ刺激后，PKCδ特异性抑制剂rottlerin不影响正常心肌细胞的凋亡

3、率，却有效抑制了轻度和中度肥大心肌细胞的凋亡。肥大心肌细胞凋亡易感性明显高于正常心肌细胞，抑制PKCδ可以抑制肥大心肌细胞凋亡，提示PKCδ参与肥大心肌细胞凋亡过程。关键词：心肌细胞；肥大；细胞凋亡；蛋白激酶Cδ；rottlerin中图分类号：R541.3;Q255ProteinkinaseCδispossiblyinvolvedinthetransitionfromhypertrophytoapoptosisofmyocardiocytesGUOWan-Gang,YUZhi-Bin*,XIEMan-JiangDepartmentofAerospacePhysiology,theFourth

4、MilitaryMedicalUniversity,Xi’an710032,ChinaAbstract:Cardiachypertrophyisanadaptiveprocesstoanincreasedhemodynamicoverload.However,theadaptionmayleadtothefragilityofmyocardiumfacingpathologicalstimuli.Inthepresentstudy,experimentsweredesignedtoexplorethesusceptibilityofhypertrophicmyocardiocytestoap

5、optoticstimuliandtheroleofproteinkinaseCδ(PKCδ)duringthetransitionfromhypertrophytoapoptosis.Endothelin-1(ET-1)-treatedcardiomyocyteswereusedasmodelofcardiachypertrophy.AngiotensinII(AngII)wasusedasanapoptoticstimulus.Cellsurfaceareawasmeasuredtodeterminetheextentofhypertrophy.Theapoptoticrateincar

6、diomyocyteswasdetectedbyHoechst33258.(1)Cellsurfaceareawasincreasedby42.5%and67.3%following1nmol/Land10nmol/LET-1treatment,respectively,ascomparedwithserum-freeculturedmyocytes.Sothemildlyandmoderatelyhypertrophicmyocytemodelsweresetup.(2)Apoptoticratesinserum-freecultured,mildlyandmoderatelyhypert

7、rophicmyocytesafterAngIItreatmentwere(15.54±1.32)%,(20.65±1.40)%and(29.33±3.52)%,respectively.Itissuggestedthathypertrophicmyocytesaremoresusceptivetoapoptoticstimulus.(3)Rottlerin,aspecificinhibitorofPKCδd