紫外线联合臭氧及高温诱导人外周血淋巴细胞凋亡作用的研究

Effect of ultraviolet light in combination with ozone and hyperthermia on apoptosis induction in human peripheral blood lymphocytes

目的:观察紫外线、臭氧及高温综合治疗对人外周血淋巴细胞凋亡水平的影响,并探讨其在免疫炎症性疾病治疗中的作用。方法:采用瑞氏-姬姆萨染色涂片镜检、琼脂糖DNA凝胶电泳及流式细胞仪定量测定技术,检测紫外线、高温及臭氧分别及联合运用时人外周血淋巴细胞的凋亡率。结果:经诱导的淋巴细胞在光镜下均可观察到典型的凋亡细胞的形态改变,DNA电泳呈现有一定间隔的梯状条带。经紫外灯照射10min、20min及30min,淋巴细胞的凋亡率依次为12.03%、16.98%和26.58%;而单纯置于温度为40、42.5及45℃的水浴中20 min,凋亡率分别为15.93%、26.83%和39.88%;经高温水浴联合紫外照射共同影响,凋亡率可达54.45%;经臭氧联合高温水浴及紫外照射共同作用,凋亡率最高达76.15%。结论:人外周血淋巴细胞在紫外线、高温及臭氧的共同影响下,其凋亡率较单一或两种因素联合作用显著提高(P<0.05),可呈现一定的时间和剂量依赖效应,以42.5℃水浴、29μg/ml臭氧联合紫外照射20 min后,淋巴细胞的凋亡率最高。

AIM：To study the effect of ultraviolet light in combination with ozone and hyperthermia on the apoptosis rate in human peripheral blood lymphocytes and to explore its mechanism in treatment of inflammatory-immune diseases.METHODS： Wright-Giemsa stain smear microscopy,DNA gel electrophoresis and flow cytometry analysis were used to detect the respective and combined apoptosis rates of ultraviolet light,ozone and hyperthermia in human peripheral blood lymphocytes.RESULTS： The induced lymphocytes showed typical morphological changes of apoptosis and DNA typical ladder type line spectrum.Each apoptosis situation was quantitatively determined by flow cytometry.After ultraviolet irradiation for 10,20 and 30 min,the apoptosis rates were,respectively,12.03%,16.98% and 26.58%.After the high temperature at 40,42.5 and 45℃ for 20 min,the apoptosis rates were,respectively,15.9%,26.83% and 39.88%.After ultraviolet irradiation and the high temperature,the apoptosis rate was 54.5%.After ultraviolet irradiation combined with hyperthermia and ozone,the highest apoptosis rate reached up to 76.2%.CONCLUSION： Lymphocyte apoptosis rates induced by ultraviolet light combined with ozone and hyperthermia increase significantly.Apoptosis rates of lymphocytes increase in a dose and time-dependent fashion within a certain range and reach the highest point after ultraviolet irradiation combined with hyperthermia at 42.5 C and ozone concentrations on 29 μg/ml for 20 min.