中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
13期
1024-1028
,共5页
王湘芸%吴莹%钱玲玲%党时鹏%季圆%徐凤%王萌%汤徐%姚勇
王湘蕓%吳瑩%錢玲玲%黨時鵬%季圓%徐鳳%王萌%湯徐%姚勇
왕상예%오형%전령령%당시붕%계원%서봉%왕맹%탕서%요용
腺苷三磷酸%大电导钙激活钾通道%钙离子%冠状动脉%平滑肌细胞
腺苷三燐痠%大電導鈣激活鉀通道%鈣離子%冠狀動脈%平滑肌細胞
선감삼린산%대전도개격활갑통도%개리자%관상동맥%평활기세포
Adenosine triphosphate%Large conductance calcium activated potassium channel%Calcium%Coronary artery%Smooth muscle cell
目的 探讨腺苷三磷酸(ATP)对冠状动脉平滑肌细胞大电导钙激活钾通道(BK通道)的激活作用及其机制.方法 采用酶消化法急性分离正常大鼠冠状动脉平滑肌细胞;膜片钳实验技术记录全细胞状态下BK通道电流;钙离子成像技术记录ATP对细胞内钙离子浓度影响.结果 膜片钳实验结果显示,加入1 mmol/L ATP前后,BK通道电流密度分别为(137±13) pA/pF和(179±15)pA/pF(P ＜0.05);钙离子成像结果显示,加入0.5 mmol/L ATP前后,细胞内钙离子浓度荧光信号强度比值分别为2.46±0.08和4.04±0.21 (P＜0.05).分别采用嘌呤(P2Y1)受体阻滞剂MRS2179、磷脂酶C(PLC)受体阻滞剂U73122和三磷酸肌醇(IP3)受体阻滞剂2-APB孵育冠状动脉平滑肌细胞,再测定加入0.5 mmol/L ATP后3组细胞内钙离子浓度荧光信号强度比值,分别是2.70±0.06、2.65±0.12和2.69±0.13,与未加入3种阻滞剂相比,钙离子浓度荧光信号强度比值均明显降低(P＜0.05).结论 ATP可以通过P2Y1-PLC-IP3通路升高细胞内钙离子浓度,从而激活BK通道.
目的 探討腺苷三燐痠(ATP)對冠狀動脈平滑肌細胞大電導鈣激活鉀通道(BK通道)的激活作用及其機製.方法 採用酶消化法急性分離正常大鼠冠狀動脈平滑肌細胞;膜片鉗實驗技術記錄全細胞狀態下BK通道電流;鈣離子成像技術記錄ATP對細胞內鈣離子濃度影響.結果 膜片鉗實驗結果顯示,加入1 mmol/L ATP前後,BK通道電流密度分彆為(137±13) pA/pF和(179±15)pA/pF(P ＜0.05);鈣離子成像結果顯示,加入0.5 mmol/L ATP前後,細胞內鈣離子濃度熒光信號彊度比值分彆為2.46±0.08和4.04±0.21 (P＜0.05).分彆採用嘌呤(P2Y1)受體阻滯劑MRS2179、燐脂酶C(PLC)受體阻滯劑U73122和三燐痠肌醇(IP3)受體阻滯劑2-APB孵育冠狀動脈平滑肌細胞,再測定加入0.5 mmol/L ATP後3組細胞內鈣離子濃度熒光信號彊度比值,分彆是2.70±0.06、2.65±0.12和2.69±0.13,與未加入3種阻滯劑相比,鈣離子濃度熒光信號彊度比值均明顯降低(P＜0.05).結論 ATP可以通過P2Y1-PLC-IP3通路升高細胞內鈣離子濃度,從而激活BK通道.
목적 탐토선감삼린산(ATP)대관상동맥평활기세포대전도개격활갑통도(BK통도)적격활작용급기궤제.방법 채용매소화법급성분리정상대서관상동맥평활기세포;막편겸실험기술기록전세포상태하BK통도전류;개리자성상기술기록ATP대세포내개리자농도영향.결과 막편겸실험결과현시,가입1 mmol/L ATP전후,BK통도전류밀도분별위(137±13) pA/pF화(179±15)pA/pF(P ＜0.05);개리자성상결과현시,가입0.5 mmol/L ATP전후,세포내개리자농도형광신호강도비치분별위2.46±0.08화4.04±0.21 (P＜0.05).분별채용표령(P2Y1)수체조체제MRS2179、린지매C(PLC)수체조체제U73122화삼린산기순(IP3)수체조체제2-APB부육관상동맥평활기세포,재측정가입0.5 mmol/L ATP후3조세포내개리자농도형광신호강도비치,분별시2.70±0.06、2.65±0.12화2.69±0.13,여미가입3충조체제상비,개리자농도형광신호강도비치균명현강저(P＜0.05).결론 ATP가이통과P2Y1-PLC-IP3통로승고세포내개리자농도,종이격활BK통도.
Objective To explore the effects and activation mechanism of adenosine triphosphate (ATP) on large conductance calcium-activated potassium channel (BK channel) in coronary artery smooth muscle cells.Methods Coronary smooth muscle cells were isolated by enzyme digestion from SpragueDawley rats.And BK currents were recorded by patch clamp technique in whole cell configuration.The effects of ATP on cytosolic calcium concentrations were examined by recording the changes of fluorescence intensity ratios.Results BK current densities were (137 ± 13) pA/pF and (179 ± 15) pA/pF before and after a perfusion of 1 mmol/L ATP (P ＜ 0.05).The fluorescence ratios were 2.46 ± 0.08 and 4.04 ± 0.21 (P ＜ 0.05) before and after 0.5 mmol/L perfusion.After incubating with purine receptor (P2Y1) blocker MRS2179,phospholipase C (PLC) blocker U73122 and inositol triphosphate (IP3) blocker 2-APB,the fluorescence ratios were 2.7 ± 0.06,2.65 ± 0.12 and 2.69 ± 0.13 respectively.Compared with control group,all fluorescence ratios decreased after incubating with three blockers (P ＜ 0.05).Conclusion ATP may elevate intracellular calcium concentration via P2Y1-PLC-IP3 pathway consequently activating BK channel.
