CAJ | 학술논문

目的研究电磁脉冲(electromagnetic pulse,EMP)、S波段高功率微波(S-band high power microwave,S-HPM)、X波段高功率微波(X-band high power microwave,X-HPM)3种不同波段电磁辐射对Raji细胞内Ca2+浓度、Caspase-3蛋白表达的影响及两者之间的相互关系,探讨电磁辐射损伤的相关调控机制.方法常规培养Raji细胞,用S-HPM、X-HPM和EMP 3种不同波段的电磁辐射照射Raji细胞,设假照射为对照组.于对数生长期照射,照射后6h收集细胞,采用激光扫描共聚焦显微镜检测细胞内Ca2+浓度的改变,采用免疫印迹法(Western blot)检测细胞内Caspase-3蛋白的表达.结果 3种不同波段的电磁辐射照射后6h,Raji细胞内Ca2+浓度增加(EMP组、S-HPM组、X-HPM组Ca2+荧光强度分别为69.56±1.71、50.06±1.89、70.68±1.59),Caspase-3蛋白表达水平上调(EMP组、S-HPM组、X-HPM组Caspase-3蛋白表达水平分别为0.964±0.12、0.586±0.16、0.970±0.07),各辐射组Raji细胞Ca2+荧光强度、Caspase-3蛋白表达水平均明显高于对照组(分别为43.08±2.08、0.444 ±0.13),差异均有统计学意义(P＜0.01)；EMP组和X-HPM组Ca2+荧光强度、Caspase-3蛋白表达水平均明显高于S-HPM组,差异亦有统计学意义(P＜0.01):EMP与X-HPM组间的差异无统计学意义(P＞0.05),但均较S-HPM组上调明显,差异均有统计学意义(P＜0.01).辐射组Ca2+浓度增加的规律和Caspase-3蛋白表达上调的规律具有一致性,线性回归分析结果显示,Caspase-3蛋白的表达随着Ca2+浓度的增高而上调,二者具有一定的相关性(P＜0.01).结论 3种辐射损伤效应可能是通过增加细胞内Ca2+浓度进一步诱导Caspase-3过表达所致.
목적 연구전자맥충(electromagnetic pulse,EMP)、S파단고공솔미파(S-band high power microwave,S-HPM)、X파단고공솔미파(X-band high power microwave,X-HPM)3충불동파단전자복사대Raji세포내Ca2+농도、Caspase-3단백표체적영향급량자지간적상호관계,탐토전자복사손상적상관조공궤제.방법 상규배양Raji세포,용S-HPM、X-HPM화EMP 3충불동파단적전자복사조사Raji세포,설가조사위대조조.우대수생장기조사,조사후6h수집세포,채용격광소묘공취초현미경검측세포내Ca2+농도적개변,채용면역인적법(Western blot)검측세포내Caspase-3단백적표체.결과 3충불동파단적전자복사조사후6h,Raji세포내Ca2+농도증가(EMP조、S-HPM조、X-HPM조Ca2+형광강도분별위69.56±1.71、50.06±1.89、70.68±1.59),Caspase-3단백표체수평상조(EMP조、S-HPM조、X-HPM조Caspase-3단백표체수평분별위0.964±0.12、0.586±0.16、0.970±0.07),각복사조Raji세포Ca2+형광강도、Caspase-3단백표체수평균명현고우대조조(분별위43.08±2.08、0.444 ±0.13),차이균유통계학의의(P＜0.01)；EMP조화X-HPM조Ca2+형광강도、Caspase-3단백표체수평균명현고우S-HPM조,차이역유통계학의의(P＜0.01):EMP여X-HPM조간적차이무통계학의의(P＞0.05),단균교S-HPM조상조명현,차이균유통계학의의(P＜0.01).복사조Ca2+농도증가적규률화Caspase-3단백표체상조적규률구유일치성,선성회귀분석결과현시,Caspase-3단백적표체수착Ca2+농도적증고이상조,이자구유일정적상관성(P＜0.01).결론 3충복사손상효응가능시통과증가세포내Ca2+농도진일보유도Caspase-3과표체소치.
Objective To study the effects of electromagnetic pulse (EMP),S-band high power microwave (S-HPM),and X-band high power microwave (X-HPM) on the Ca2+ concentration and caspase-3 expression in Raji cells and the rclationship between Ca2+ concentration and caspase-3 expression,and to investigate the regulatory mechanism of electromagnetic radiation damage.Methods Raji cells were cultured conventionally.Some cells were irradiated by EMP,S-HPM,and X-HPM in the logarithmic growth phase for 6 hours and then collected; others received sham irradiation as a control.The Ca2+ concentration in the cells was measured by laser scanning confocal microscope; the caspase-3 expression in the cells was evaluated by Western blot.Results Compared with the control group (Ca2+ fluorescence intensity =43.08±2.08; caspase-3 expression level=0.444±0.13),the EMP,S-HPM,and X-HPM groups had significantly increased Ca2+ concentrations,with Ca2+fluorescence intensities of 69.56±1.71,50.06±1.89,and 70.68±1.59,respectively (P＜0.01),and had upregulated caspase-3 expression,with expression levels of 0.964±0.12,0.586±0.16,and 0.970±0.07,respectively (P＜0.01).Each of the EMP and X-HPM groups had significantly higher Ca2+ fluorescence intensity and caspase-3 expression level than the S-HPM group (P＜0.01),but there were no significant differences between the EMP and X-HPM groups.The linear regression analysis showed that the caspase-3 expression was upregulated as the Ca2+ concentration increased,with a positive correlation between them (P＜0.01).Conclusion EMP,S-HPM,and X-HPM cause damage probably by increasing the Ca2+ concentration in cells and in turn inducing caspase-3 overexpression.