中华老年医学杂志
中華老年醫學雜誌
중화노년의학잡지
Chinese Journal of Geriatrics
2008年
12期
927-930
,共4页
周永明%黎明江%丘天翼%唐艳红%黄从新
週永明%黎明江%丘天翼%唐豔紅%黃從新
주영명%려명강%구천익%당염홍%황종신
TRPM阳离子通道%成纤维细胞%心肌梗死%RNA,小分子干扰
TRPM暘離子通道%成纖維細胞%心肌梗死%RNA,小分子榦擾
TRPM양리자통도%성섬유세포%심기경사%RNA,소분자간우
TRPM cation channels%Fibroblasts%Myocardial infarction%SiRNA
目的 观察小鼠心肌成纤维细胞(CFs)在心肌梗死后瞬时受体电位通道亚族M7(TRPM7)样电流的变化及其对胶原生成的影响,探讨TRPM7离子通道在心脏纤维化形成中的潜在病理生理作用. 方法 (1)制备小鼠心肌梗死模型并分离CFs细胞;(2)培养传代CFs及小分子干扰RNA(SiRNA)技术感染;(3)应用膜片钳技术观察CFs缺血后TRPM7通道内外向电流特征;(4)钙荧光显像技术观察缺血心肌对CFs的钙离子内流影响;(5)测定缺血对CFs总胶原含量的影响.结果 (1)心肌缺血能使CFs的含钙离子内向电流较对照组显著增加,分别为(7.4±0.7)pA/pF和(16.25±1.7)pA/pF(P<0.05);(2)SiRNA使TRPM7样电流的mRNA水平明显下降,且电流幅值减少约90%;(3)心肌缺血组CFs的TRPM的mRNA丰度及总胶原含量较基础值增加2.3倍左右.结论 致纤维化因子心肌缺血可通过TRPM7样电流介导的钙离子信号机制调节CFs功能,在心肌组织纤维化的病理生理过程中发挥重要作用.
目的 觀察小鼠心肌成纖維細胞(CFs)在心肌梗死後瞬時受體電位通道亞族M7(TRPM7)樣電流的變化及其對膠原生成的影響,探討TRPM7離子通道在心髒纖維化形成中的潛在病理生理作用. 方法 (1)製備小鼠心肌梗死模型併分離CFs細胞;(2)培養傳代CFs及小分子榦擾RNA(SiRNA)技術感染;(3)應用膜片鉗技術觀察CFs缺血後TRPM7通道內外嚮電流特徵;(4)鈣熒光顯像技術觀察缺血心肌對CFs的鈣離子內流影響;(5)測定缺血對CFs總膠原含量的影響.結果 (1)心肌缺血能使CFs的含鈣離子內嚮電流較對照組顯著增加,分彆為(7.4±0.7)pA/pF和(16.25±1.7)pA/pF(P<0.05);(2)SiRNA使TRPM7樣電流的mRNA水平明顯下降,且電流幅值減少約90%;(3)心肌缺血組CFs的TRPM的mRNA豐度及總膠原含量較基礎值增加2.3倍左右.結論 緻纖維化因子心肌缺血可通過TRPM7樣電流介導的鈣離子信號機製調節CFs功能,在心肌組織纖維化的病理生理過程中髮揮重要作用.
목적 관찰소서심기성섬유세포(CFs)재심기경사후순시수체전위통도아족M7(TRPM7)양전류적변화급기대효원생성적영향,탐토TRPM7리자통도재심장섬유화형성중적잠재병리생리작용. 방법 (1)제비소서심기경사모형병분리CFs세포;(2)배양전대CFs급소분자간우RNA(SiRNA)기술감염;(3)응용막편겸기술관찰CFs결혈후TRPM7통도내외향전류특정;(4)개형광현상기술관찰결혈심기대CFs적개리자내류영향;(5)측정결혈대CFs총효원함량적영향.결과 (1)심기결혈능사CFs적함개리자내향전류교대조조현저증가,분별위(7.4±0.7)pA/pF화(16.25±1.7)pA/pF(P<0.05);(2)SiRNA사TRPM7양전류적mRNA수평명현하강,차전류폭치감소약90%;(3)심기결혈조CFs적TRPM적mRNA봉도급총효원함량교기출치증가2.3배좌우.결론 치섬유화인자심기결혈가통과TRPM7양전류개도적개리자신호궤제조절CFs공능,재심기조직섬유화적병리생리과정중발휘중요작용.
Objective To investigate the changes of TRPM 7-like current in mouse cardiac fibroblast(CFs) after myocardial infarction and the effect of myocardial ischemia on the TRPM 7 expression and current. Methods (1) The model of myocardial infarction was made and CFs were isolated;(2) CFs were cultured and infected by TRPM 7 siRNA;(3) The effects of myocardial ischemia on TRPM 7 current were recorded by whole cell patch clamp technique;(4) The influences of myocardial isehemia on Ca2+ influx in CFs were recorded by Ca2+ fluorescence imaging. (5) The effects of ischemia on total collagen content of CFs were studied. Results (1) Ca2+ inward current of CFs was increased after myocardial infarction [(16.2±1.7) vs. (7.4±0.7) pA/pF, P<0.053];(2) TRPM 7 current was reduced by 90% after siRNA infection;(3) The total collagen content of CFs after ischemia was approximately 2.3-fold higher than basic value. Conclusions Ca2+ influx in CFs plays an important role in the pathophysiological process of myocardial fibrosis.
