诱导恒河猴外周血淋巴细胞凋亡的研究  

Apoptosis induction of rhesus peripheral blood lymphocytes

作　　者：范礼佩[1] 孙尔维[1] 袁小澎[1] 卢晓[2] 王海琴[1] 刘占国 李民[1] 赵明[1] 钟世镇[3] 

机构地区：[1]南方医科大学珠江医院器官移植科,广东广州510282 [2]南方医科大学免疫教研室,广东广州510515 [3]南方医科大学解剖教研室,广东广州510515

出　　处：《南方医科大学学报》2007年第5期628-630,共3页Journal of Southern Medical University

基　　金：国家自然科学基金(30571757);广东省自然科学基金团队项目~~

摘　　要：目的研究诱导恒河猴外周血淋巴细胞凋亡的方法。方法采用高能X线、γ射线和紫外线照射法,诱导恒河猴外周血淋巴细胞凋亡。收集处理后的细胞,应用PI和FITCAnnexin-V进行标记,流式细胞计数仪检测。结果高能X线和γ射线照射恒河猴外周血淋巴细胞,诱导的凋亡率低;紫外线照射恒河猴外周血淋巴细胞诱导的凋亡率可达60%。小牛血清的浓度为10%,照射距离为20cm,照射60min时,其早期凋亡率为58.85%,晚期凋亡率为11.5%。凋亡细胞随剂量的增加及时间推移而增加,并表现出时效和量效关系。结论诱导恒河猴外周血淋巴细胞凋亡,紫外线法比高能X线和γ射线法敏感,效果好。紫外线照射的最佳条件是10%小牛血清的1640培养基,照射距离20cm,照射时间60min。Objective To establish a method for inducing apoptosis of rhesus peripheral blood lymphocytes （PBLs）. Methods Rhesus PBLs were irradiated with X-ray, ^60Co gamma-rays and ultraviolet （UVC254 ran）, respectively, and the cell apoptosis was evaluated with flow cytometry using annexin-V staining and propidium iodide staining. Results X-ray and ^60Co gamma-ray irradiation induced only low apoptotic rates of the PBLs, and UVC resulted in the highest apoptotic rate of about 60%. UVC irradiation of the PBLs in RPMI supplemented with 10% heat-inactivated fetal calf serum for 60 min at a distance of 20 cm led to an early apoptotic rate of 58.85% and necrotic rate of 11.5%. The apoptotic rate of PBLs increased in a doseand time-dependent fashion. Conclusion For inducing apoptosis of the rhesus PBLs, UVC can be more effective than X-ray and ^60Co gamma-ray. The highest apoptotic rate can be achieved when the rhesus PBLs in RPMI supplemented with 10% heat-inactivated fetal calf serum are exposed to UVC for 60 min at the distance of 20 cm.

关 键 词：恒河猴 淋巴细胞 X线 Γ射线 紫外线 凋亡 

分 类 号：R392[医药卫生—免疫学]