CAJ | 학술논문

目的探讨γH2AX识别抗体流式细胞术(FCM)检测1,2-二氯乙烷(1,2-DCE)接触工人外周血淋巴细胞DNA损伤的可行性及1,2-DCE对健康人外周血淋巴细胞DNA的损伤.方法提取某制鞋厂接触1,2-DCE的工人21例(接触组)和该厂未接触1,2-DCE的工人27例(内对照组)及某海岛非职业接触有害因素居民28例(外对照组)的外周血淋巴细胞,采用FCM法检测淋巴细胞DNA损伤情况;用不同浓度(5、10、20和30 μmol/L)的1,2-DCE分别对健康人外周血淋巴细胞体外染毒0.5、1.0 h,采用FCM法检测淋巴细胞DNA损伤情况.结果接触组工人DNA损伤率(4.05%±2.55%)明显高于内对照组工人(1.97%±1.40%)和外对照组人群(0.23%±0.13%),内对照组明显高于外对照组,差异均有统计学意义(P＜0.01或P＜0.05);接触组工人外周血淋巴细胞的几何平均荧光强度(3.33±3.01)与内对照组(2.07±0.58)比较,差异有统计学意义(P＜0.05).接触组不同工龄工人DNA损伤率和外周血淋巴细胞的几何平均荧光强度比较,差异均无统计学意义(P＞0.05).体外染毒0.5 h时,20、30μmol/L染毒组外周血淋巴细胞的几何平均荧光强度与阴性对照组比较,差异有统计学意义(P＜0.01).体外染毒1.0 h时,20、30μmol/L染毒组的DNA损伤率与阴性对照组比较,差异有统计学意义(P＜0.05,P＜0.01),10、20、30 μmol/L染毒组外周血淋巴细胞的几何平均荧光强度与阴性对照组比较,差异均有统计学意义(P＜0.05,P＜0.01).结论 1,2-DCE具有遗传损伤作用,FCM法是一种检测外周血淋巴细胞DNA损伤的有效方法.
목적 탐토γH2AX식별항체류식세포술(FCM)검측1,2-이록을완(1,2-DCE)접촉공인외주혈림파세포DNA손상적가행성급1,2-DCE대건강인외주혈림파세포DNA적손상.방법 제취모제혜엄접촉1,2-DCE적공인21례(접촉조)화해엄미접촉1,2-DCE적공인27례(내대조조)급모해도비직업접촉유해인소거민28례(외대조조)적외주혈림파세포,채용FCM법검측림파세포DNA손상정황;용불동농도(5、10、20화30 μmol/L)적1,2-DCE분별대건강인외주혈림파세포체외염독0.5、1.0 h,채용FCM법검측림파세포DNA손상정황.결과 접촉조공인DNA손상솔(4.05%±2.55%)명현고우내대조조공인(1.97%±1.40%)화외대조조인군(0.23%±0.13%),내대조조명현고우외대조조,차이균유통계학의의(P＜0.01혹P＜0.05);접촉조공인외주혈림파세포적궤하평균형광강도(3.33±3.01)여내대조조(2.07±0.58)비교,차이유통계학의의(P＜0.05).접촉조불동공령공인DNA손상솔화외주혈림파세포적궤하평균형광강도비교,차이균무통계학의의(P＞0.05).체외염독0.5 h시,20、30μmol/L염독조외주혈림파세포적궤하평균형광강도여음성대조조비교,차이유통계학의의(P＜0.01).체외염독1.0 h시,20、30μmol/L염독조적DNA손상솔여음성대조조비교,차이유통계학의의(P＜0.05,P＜0.01),10、20、30 μmol/L염독조외주혈림파세포적궤하평균형광강도여음성대조조비교,차이균유통계학의의(P＜0.05,P＜0.01).결론 1,2-DCE구유유전손상작용,FCM법시일충검측외주혈림파세포DNA손상적유효방법.
Objective To study DNA damage of human peripheral blood lymphocytes exposed to 1,2dichloroethane (1,2-DCE) with flow cytometry (FCM) assay. Methods The lymphocytes were obtained from 21 workers who are occupationally exposed to 1, 2-DCE (exposed group ) and 27 workers who were not exposed to 1,2-DCE in the same factory (inner control ) and 28 island residents who had never been occupationally exposed to adverse factors (external control ). FCM assay was adopted to detect DNA damage of the lymphocvtes of each group. Lymphocytes of the health people were incubated with 1 ,2 - DCE at different doses, and FCM assay was used to detect DNA damage. Results DNA damage rate (%) of the exposed group of exposed workers (4.05%±2.55%) was significantly higher than the inner control group of workers ( 1.97%± 1.40% ) and external control groups of island residents (0.23%±0.13%), and the DNA damage of inner control was higher than the external control, all the differences were statistically significant (P＜0.01 or P＜0.05 ).The geometric mean fluorescence intensity of the workers in the exposed group (3.33±3.01) was significantly higher than the (2.07±0.58) only (P＜0.05). There was no significant difference in the DNA damage rate as well as the geometric mean fluorescence intensity among the exposed group of workers with different years of working period (P＞0.05). In vitro, the fluorescence intensity at the dose of 20,30 μmol/L for 0.5 h exposure showed statistical significance compared with the negative control group (P＜0.01). The DNA damage rate at the dose of 20, 30μmol/L for 1.0 h exposure was statistically significant compared with the negative control group (P＜0.05,P＜0.01 ); The fluorescence intensity at the dose of 10,20,30 μmol/L for 1.0 h exposure was statistically significant compared with the negative control group(P＜0.05,P＜0.01 ). Conclusion 1 ,2-DEC can cause DNA damage. And γH2AX FCM assay can be a sensitive, objective and effective method of detecting DNA damage of peripheral blood lymphocytes.