中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2011年
6期
642-645
,共4页
王丽娜%郭津%吴博%高秀香
王麗娜%郭津%吳博%高秀香
왕려나%곽진%오박%고수향
受体,钙敏感%蛋白激酶C%血管紧张素Ⅱ%心肌肥大
受體,鈣敏感%蛋白激酶C%血管緊張素Ⅱ%心肌肥大
수체,개민감%단백격매C%혈관긴장소Ⅱ%심기비대
Receptors,calcium-sensing%Protein kinase C%Angiotensin Ⅱ%Cardiac hypertrophy
目的 观察钙敏感受体(CaSR)在血管紧张素Ⅱ(Ang Ⅱ)诱导心肌细胞肥大中的作用和可能机制.方法 用Ang Ⅱ处理原代新生大鼠心室肌细胞复制心肌肥大细胞模型；用CaSR激动剂氯化钆(GdCl3),GdCl3+蛋白激酶C(PKC)通路阻断剂(Ro318220)处理AngⅡ诱导的肥大心肌细胞分别作为GdCl3、Ro318220组.通过苏木素-伊红染色(HE)法测定细胞直径,考马斯亮蓝蛋白试剂盒测定蛋白含量来评价细胞肥大的情况；利用激光共聚焦显微镜检测心肌细胞内钙浓度([Ca2+]i；Western blot法检测CaSR和PKC通路的蛋白表达.结果 ①与对照组(0.1263±0.0443)比较,Ang Ⅱ组(0.1963±0.0375)和GdCl3组(0.2778±0.0564)CaSR蛋白表达明显增加(P均＜ 0.05),且GdCl3组明显高于AngⅡ组(P＜0.05).②与对照组(222.70±22.09)比较,Ang Ⅱ组(392.16±36.85)和GdCl3组(502.60±44.21)心肌细胞内[Ca2+]i显著增加(P均＜0.05)；与Ang Ⅱ组比较,GdCl3组[Ca2+]i显著增加(P＜0.05).③与对照组比较,Ang Ⅱ可诱导心肌细胞肥大,GdCl3可促进Ang Ⅱ的诱导作用,而Ro318220可抑制GdCl3的作用；④与对照组(0.27±0.07、0.69±0.06、0.87±0.04)比较,Ang Ⅱ组PKCα、PKCε和PKCδ蛋白表达明显增加(0.60±0.16、1.02±0.13、1.20±0.18,P均＜0.05),GdCl3组PKCα、PKCε蛋白表达明显增加(0.82±0.16、1.34±0.12,P均＜0.05)；与Ang Ⅱ组比较,GdCl3组PKCα、PKCε蛋白表达明显增加(P均＜ 0.05)；与GdCl3组比较,Ro318220组PKCα、PKCε蛋白表达(0.41±0.10、0.85±0.14)明显减少(P均＜ 0.05).结论 PKC通路参与CaSR激活促进心肌细胞肥大的信号转导.
目的 觀察鈣敏感受體(CaSR)在血管緊張素Ⅱ(Ang Ⅱ)誘導心肌細胞肥大中的作用和可能機製.方法 用Ang Ⅱ處理原代新生大鼠心室肌細胞複製心肌肥大細胞模型；用CaSR激動劑氯化釓(GdCl3),GdCl3+蛋白激酶C(PKC)通路阻斷劑(Ro318220)處理AngⅡ誘導的肥大心肌細胞分彆作為GdCl3、Ro318220組.通過囌木素-伊紅染色(HE)法測定細胞直徑,攷馬斯亮藍蛋白試劑盒測定蛋白含量來評價細胞肥大的情況；利用激光共聚焦顯微鏡檢測心肌細胞內鈣濃度([Ca2+]i；Western blot法檢測CaSR和PKC通路的蛋白錶達.結果 ①與對照組(0.1263±0.0443)比較,Ang Ⅱ組(0.1963±0.0375)和GdCl3組(0.2778±0.0564)CaSR蛋白錶達明顯增加(P均＜ 0.05),且GdCl3組明顯高于AngⅡ組(P＜0.05).②與對照組(222.70±22.09)比較,Ang Ⅱ組(392.16±36.85)和GdCl3組(502.60±44.21)心肌細胞內[Ca2+]i顯著增加(P均＜0.05)；與Ang Ⅱ組比較,GdCl3組[Ca2+]i顯著增加(P＜0.05).③與對照組比較,Ang Ⅱ可誘導心肌細胞肥大,GdCl3可促進Ang Ⅱ的誘導作用,而Ro318220可抑製GdCl3的作用；④與對照組(0.27±0.07、0.69±0.06、0.87±0.04)比較,Ang Ⅱ組PKCα、PKCε和PKCδ蛋白錶達明顯增加(0.60±0.16、1.02±0.13、1.20±0.18,P均＜0.05),GdCl3組PKCα、PKCε蛋白錶達明顯增加(0.82±0.16、1.34±0.12,P均＜0.05)；與Ang Ⅱ組比較,GdCl3組PKCα、PKCε蛋白錶達明顯增加(P均＜ 0.05)；與GdCl3組比較,Ro318220組PKCα、PKCε蛋白錶達(0.41±0.10、0.85±0.14)明顯減少(P均＜ 0.05).結論 PKC通路參與CaSR激活促進心肌細胞肥大的信號轉導.
목적 관찰개민감수체(CaSR)재혈관긴장소Ⅱ(Ang Ⅱ)유도심기세포비대중적작용화가능궤제.방법 용Ang Ⅱ처리원대신생대서심실기세포복제심기비대세포모형；용CaSR격동제록화구(GdCl3),GdCl3+단백격매C(PKC)통로조단제(Ro318220)처리AngⅡ유도적비대심기세포분별작위GdCl3、Ro318220조.통과소목소-이홍염색(HE)법측정세포직경,고마사량람단백시제합측정단백함량래평개세포비대적정황；이용격광공취초현미경검측심기세포내개농도([Ca2+]i；Western blot법검측CaSR화PKC통로적단백표체.결과 ①여대조조(0.1263±0.0443)비교,Ang Ⅱ조(0.1963±0.0375)화GdCl3조(0.2778±0.0564)CaSR단백표체명현증가(P균＜ 0.05),차GdCl3조명현고우AngⅡ조(P＜0.05).②여대조조(222.70±22.09)비교,Ang Ⅱ조(392.16±36.85)화GdCl3조(502.60±44.21)심기세포내[Ca2+]i현저증가(P균＜0.05)；여Ang Ⅱ조비교,GdCl3조[Ca2+]i현저증가(P＜0.05).③여대조조비교,Ang Ⅱ가유도심기세포비대,GdCl3가촉진Ang Ⅱ적유도작용,이Ro318220가억제GdCl3적작용；④여대조조(0.27±0.07、0.69±0.06、0.87±0.04)비교,Ang Ⅱ조PKCα、PKCε화PKCδ단백표체명현증가(0.60±0.16、1.02±0.13、1.20±0.18,P균＜0.05),GdCl3조PKCα、PKCε단백표체명현증가(0.82±0.16、1.34±0.12,P균＜0.05)；여Ang Ⅱ조비교,GdCl3조PKCα、PKCε단백표체명현증가(P균＜ 0.05)；여GdCl3조비교,Ro318220조PKCα、PKCε단백표체(0.41±0.10、0.85±0.14)명현감소(P균＜ 0.05).결론 PKC통로삼여CaSR격활촉진심기세포비대적신호전도.
Objective To explore the roles and possible mechanism of calcium-sensing receptor(CaSR) in cell cardiac hypertrophy model using angiotensin Ⅱ (Ang Ⅱ ).Methods The cultured neonatal rat ventricular myocytes were treated with Ang Ⅱ as cell cardiac hypertrophy model.Hypertrophic neonatal rat cardiomyocytes were treated with GdCl3(a specific agonist of CaSR) and/or with Ro318220(a specific inhibitor of PKC pathway).To evaluate the status of cardiac hypertrophy,cell diameter was observed by HE dyeing,and protein content was determined through coomassie brilliant blue protein kit.The intracellular calcium concentration( [ Ca2+]i) was determined by laser scanning confocal microscope.The protein expression of CaSR and PKC pathway were analyzed using Western blotting.Results ①Compared to the control group(0.1263 ± 0.0443),the protein expression of CaSR was increased in Ang Ⅱ group and in GdCl3 group(0.1963 ± 0.0375,0.2778 ± 0.0564,all P＜ 0.05).Moreover,compared with Ang Ⅱ alone,the increase was significant in GdCl3 group(P ＜ 0.05).②Compared to control group(222.70 ± 22.09),AngⅡ group(392.16 ± 36.85) remarkably increased [Ca2+]i(P＜ 0.05),and this increase of [Ca2+]i was further enhanced in GdCl3 group (502.60 ± 44.21) versus Ang Ⅱ group (P ＜ 0.05).③Compared to control group,Ang Ⅱ could induce cardiomyocyte hypertrophy,and GdCl3 enhanced the effect.Moreover,this enhancement was attenuated by Ro318220.④Compared to control group(0.27 ± 0.07,0.69 ± 0.06,0.87 ± 0.04),the protein expression of PKCα,PKCε and PKCδ was increased in Ang Ⅱ group(0.60 ± 0.16,1.02 ± 0.13,1.20 ± 0.18,all P＜ 0.05) and the protein expression of PKCα,PKCε was increased in GdCl3 group(0.82 ± 0.16,1.34 ± 0.12,all P ＜ 0.05).Moreover,compared with Ang Ⅱ group,the protein expression of PKCα,PKCε was obviously increased in GdCl3 group (all P ＜ 0.05);compared with GdCl3 group,the protein expression of PKCα,PKCε(0.41 ± 0.10,0.85 ± 0.14) was obviously decreased in Ro318220 group(all P ＜ 0.05).Conclusions CaSR is involved in cardiac hypertrophy induced by Ang Ⅱ through PKC pathway in cultured neonatal rat cardiomyocytes.
