心电与循环
心電與循環
심전여순배
Journal of Electrocardiology(China)
2013年
3期
188-192
,共5页
心肌成纤维细胞%血管紧张素Ⅱ%电场刺激%钙-钙调素依赖性蛋白激酶Ⅱ
心肌成纖維細胞%血管緊張素Ⅱ%電場刺激%鈣-鈣調素依賴性蛋白激酶Ⅱ
심기성섬유세포%혈관긴장소Ⅱ%전장자격%개-개조소의뢰성단백격매Ⅱ
Cardiac fibroblasts%Angiotensin II%Electrical field stimulation%Calcium calmodulin depen-dent protein Kinase II
目的探讨钙-钙调素依赖性蛋白激酶(CaMK)Ⅱ对大鼠心肌成纤维细胞增殖的影响及其机制.方法培养新生SD大鼠心肌成纤维细胞,分为正常对照组,AngⅡ(0.1μmol/L)组,AngⅡ+AIP(0.2μmol/L)组,AngⅡ+AIP(0.5μmol/L)组,AngⅡ+AIP(1.0μmol/L)组;EFS(10V1.0Hz)组,EFS+AIP(0.2μmol/L)组,EFS+AIP(0.5μmol/L)组,EFS+AIP (1.0μmol/L)组;MTT法及细胞记数法分别测定心肌成纤维细胞增殖;免疫印记法检测CaMKⅡδB的蛋白表达;RT-PCR检测CaMKⅡδB、C mRNA水平.结果 CaMKⅡ抑制剂能抑制AngⅡ或EFS诱导的心肌成纤维细胞增殖(P<0.05);抑制AngⅡ引起的CaMKⅡδB、δC表达增加(P<0.05);AngⅡ刺激24h CaMKⅡ活性升高(P<0.05),48h升高更明显(P<0.01).当电压从10V增加至40V,CaMKⅡ活性不断增加,呈电压依赖(P<0.05).结论 CaMKⅡ阻断剂对心肌成纤维细胞增殖有抑制作用,可能与抑制CaMKⅡδB和δC表达有关.
目的探討鈣-鈣調素依賴性蛋白激酶(CaMK)Ⅱ對大鼠心肌成纖維細胞增殖的影響及其機製.方法培養新生SD大鼠心肌成纖維細胞,分為正常對照組,AngⅡ(0.1μmol/L)組,AngⅡ+AIP(0.2μmol/L)組,AngⅡ+AIP(0.5μmol/L)組,AngⅡ+AIP(1.0μmol/L)組;EFS(10V1.0Hz)組,EFS+AIP(0.2μmol/L)組,EFS+AIP(0.5μmol/L)組,EFS+AIP (1.0μmol/L)組;MTT法及細胞記數法分彆測定心肌成纖維細胞增殖;免疫印記法檢測CaMKⅡδB的蛋白錶達;RT-PCR檢測CaMKⅡδB、C mRNA水平.結果 CaMKⅡ抑製劑能抑製AngⅡ或EFS誘導的心肌成纖維細胞增殖(P<0.05);抑製AngⅡ引起的CaMKⅡδB、δC錶達增加(P<0.05);AngⅡ刺激24h CaMKⅡ活性升高(P<0.05),48h升高更明顯(P<0.01).噹電壓從10V增加至40V,CaMKⅡ活性不斷增加,呈電壓依賴(P<0.05).結論 CaMKⅡ阻斷劑對心肌成纖維細胞增殖有抑製作用,可能與抑製CaMKⅡδB和δC錶達有關.
목적탐토개-개조소의뢰성단백격매(CaMK)Ⅱ대대서심기성섬유세포증식적영향급기궤제.방법배양신생SD대서심기성섬유세포,분위정상대조조,AngⅡ(0.1μmol/L)조,AngⅡ+AIP(0.2μmol/L)조,AngⅡ+AIP(0.5μmol/L)조,AngⅡ+AIP(1.0μmol/L)조;EFS(10V1.0Hz)조,EFS+AIP(0.2μmol/L)조,EFS+AIP(0.5μmol/L)조,EFS+AIP (1.0μmol/L)조;MTT법급세포기수법분별측정심기성섬유세포증식;면역인기법검측CaMKⅡδB적단백표체;RT-PCR검측CaMKⅡδB、C mRNA수평.결과 CaMKⅡ억제제능억제AngⅡ혹EFS유도적심기성섬유세포증식(P<0.05);억제AngⅡ인기적CaMKⅡδB、δC표체증가(P<0.05);AngⅡ자격24h CaMKⅡ활성승고(P<0.05),48h승고경명현(P<0.01).당전압종10V증가지40V,CaMKⅡ활성불단증가,정전압의뢰(P<0.05).결론 CaMKⅡ조단제대심기성섬유세포증식유억제작용,가능여억제CaMKⅡδB화δC표체유관.
Objective To study the effects of calcium calmodulin dependent protein Kinase (CaMK) Ⅱon cardiac fi-broblast proliferation of rat and its mechanisms. Methods Cardiac fibroblasts cultured from neonatal SD rat were divid-ed into nine groups as follows:control group, Ang II (0.1μmol/L)group,Ang II+AIP (0.2μmol/L)group, Ang II+AIP (0.5μmol / L)group, AngⅡ+ AIP(1.0μmol / L)group, 10V1.0Hz EFS group, EFS +0.2 μmol / L AIP group, EFS +0.5μmol/L AIP group, EFS+1.0μmol/L AIP group. Cel counting and MTT were performed to detect cardiac fibroblasts proliferation. Protein expression of CaMKII-δB was measured by Western blot. mRNA expression of CaMK II- δB and CaMK II- δC was determined by RT-PCR. Results CaMKII inhibitor prevented cardiac fibroblasts proliferation in-duced by AngⅡor EFS(P<0.05)and inhibited expression of CaMKⅡδB and δC induced by Ang II(P<0.05). The ac-tivity of CaMKII increased when being exposed to Ang II for 24 hours(P<0.05)and more significantly for 48h(P<0.01). EFS enhanced CaMKII activation in a voltage-dependent manner with the voltage from 10V to 40V(P<0.05). Conclusion CaMKII inhibitor can prevent cardiac fibroblasts proliferation by inhibiting expression of CaMKⅡδB andδC.
