CAJ | 학술논문

目的探讨流式细胞术分析静脉血管平滑肌细胞多倍体细胞的方法和意义.方法选取普通级健康成年日本大耳白兔2只,雄性,8月龄,大约3 kg;普通级SD大鼠2只,雄性,8周龄,大约200 g.人大隐静脉为2例心脏冠状动脉旁路移植术中修补血管所得.在麻醉日本大耳白兔和大鼠后开腹腔获取下腔静脉,采用多种酶组合的消化液消化兔、大鼠以及人的静脉血管得到适合流式细胞检测的静脉血管平滑肌单细胞悬液,用碘化丙啶标记细胞,利用细胞流式仪分析兔、大鼠以及人的静脉血管平滑肌单细胞悬液各5 000个细胞,检测细胞DNA含量,DNA含量翻倍的细胞为多倍体细胞.并利用荧光原位杂交技术(FISH)验证阳性对照HEK293细胞系中的多倍体的存在.结果流式结果显示分析的5 000个细胞中,阳性对照HEK293细胞的多倍体细胞为1 355个(27.1％),从2例患者中取得的大隐静脉多倍体细胞为310个(6.2％)和250个(5.0％);2只大鼠的下腔静脉平滑肌细胞中多倍体细胞为360个(7.2％)和450个(9％);2只兔的下腔静脉平滑肌细胞中多倍体细胞分别为270个(5.4％)和305个(6.1％).结论成年的静脉血管包括人大隐静脉的平滑肌细胞中普遍存在一定比例的多倍体细胞.
목적 탐토류식세포술분석정맥혈관평활기세포다배체세포적방법화의의.방법 선취보통급건강성년일본대이백토2지,웅성,8월령,대약3 kg;보통급SD대서2지,웅성,8주령,대약200 g.인대은정맥위2례심장관상동맥방로이식술중수보혈관소득.재마취일본대이백토화대서후개복강획취하강정맥,채용다충매조합적소화액소화토、대서이급인적정맥혈관득도괄합류식세포검측적정맥혈관평활기단세포현액,용전화병정표기세포,이용세포류식의분석토、대서이급인적정맥혈관평활기단세포현액각5 000개세포,검측세포DNA함량,DNA함량번배적세포위다배체세포.병이용형광원위잡교기술(FISH)험증양성대조HEK293세포계중적다배체적존재.결과 류식결과현시분석적5 000개세포중,양성대조HEK293세포적다배체세포위1 355개(27.1％),종2례환자중취득적대은정맥다배체세포위310개(6.2％)화250개(5.0％);2지대서적하강정맥평활기세포중다배체세포위360개(7.2％)화450개(9％);2지토적하강정맥평활기세포중다배체세포분별위270개(5.4％)화305개(6.1％).결론 성년적정맥혈관포괄인대은정맥적평활기세포중보편존재일정비례적다배체세포.
Objective To confirm the polyploid cells in animal and human venous vein with the use of flow cytometry.Methods The animals were 2 healthy Japanese white rabbit (male,8 months old,about 3 kg)and 2 SD rats (male,8 weeks old,about 200 g).2 human saphenous veins were from the patients who received coronary artery bypass grafting operation.The rabbits and rats were anesthetized and the abdominal cavity was opened,then the inferior vena cava was obtained.We used multiple enzymes to digest animal and human vein and to get dispersed smooth muscle cells.The DNA was stained by propidium iodide (PI),then 5 000 cells of rabbits were analyzed by flow cytometry.The confirmation of polyploid cells in tumor cell HEK293 by using Fluorescence in situ hybridization was as a positive control.Results The percentage of polyploid cells of tumor cell HEK293 was 27.1％ (1 355 cells).The percentage of polyploid cells of vascular smooth muscle cells in human great saphenous vein was 5.0％ and 6.2％,respectively.The percentage of polyploid cell in vascular smooth muscle cells of inferior vena cava in two rats were 7.2％ (360 cells)and 9％ (450 cells),respectively.The percentage of polyploid cells of vascular smooth muscle cells in two rabbits were 5.4％ (270 cells)and 6.1％ (305 cells),respectively.Conclusion Vein blood vessel contains a certain proportion of polyploid cells in vascular smooth muscle cells,including human saphenous vein.