紫外线照射对成人原代肝细胞免疫原性及蛋白合成性的影响  

Effect of ultraviolet irradiation on immunogenicity and biological activity of primary adult human hepatocytes

作　　者：邓兰 唐世刚[1] Lan Deng;Shi-Gang Tang(Department of Infectious Diseases,Peoples’Hospital of Hunan Province/First Affiliated Hospital of Hunan Normal University,Changsha 410005,Hunan Province,China)

机构地区：[1]湖南省人民医院/湖南师范大学第一附属医院感染科,湖南省长沙市410005

出　　处：《世界华人消化杂志》2020年第15期683-690,共8页World Chinese Journal of Digestology

基　　金：长沙市科技计划项目,No.kq1701048.

摘　　要：背景受体的免疫排斥反应是影响肝细胞移植疗效的主要因素,而紫外线可以引起免疫抑制的作用;寻找适当的紫外线照射强度,既能降低肝细胞免疫原性,又能避免紫外线对肝细胞的过度损伤,从而较好保存肝细胞的稳定性和细胞合成功能.目的探讨紫外线照射降低成人原代肝细胞免疫原性和对细胞生物活性的影响.方法取成人良性病变肝组织以胶原酶灌注分离肝细胞,分为对照组(0 J/m^2)及200、350、550和750 J/m^24个不同紫外线照射强度的实验组.台盼蓝拒染法和CCK-8法检测细胞活率;JC-1检测线粒体膜电位变化;混合淋巴细胞肝细胞培养(mixed lymphocyte hepatocyte culture,MLHC)检测受体T细胞增殖;并检测培养上清液中血白蛋白(serum albumin,ALB)、乳酸脱氢酶水平.结果(1)新分离的肝细胞活率大于90%;(2)CCK-8检测发现实验组200 J/m^2照射强度的肝细胞活力最高,与对照组无明显区别,明显高于其他实验组;(3)荧光显微镜观察到对照组和200 J/m^2实验组在JC-1液作用下肝细胞以红色荧光为主,随着照射强度的增大,而呈现棕色(350 J/m^2)、黄绿色(550 J/m^2)、绿色(750 J/m^2)的变化.酶标检测显示200 J/m^2组OD值最高,与对照组无显著差异,说明肝细胞膜电位稳定,细胞活性最好;随着照射强度增大,细胞膜电位随之下降,差异显著;(4)MLHC检测显示200 J/m^2照射组的肝细胞引起淋巴细胞增殖能力较对照组显著降低,而350 J/m^2、550 J/m^2、750 J/m^2组则有所增强;(5)生化检测提示200 J/m^2组的ALB水平最高,与对照组无差别.培养第3天肝细胞的分泌和合成功能处于最佳状态.结论强度200 J/m^2紫外线照射可降低成人原代肝细胞引起T细胞增殖能力;肝细胞的活力和合成功能得到较好保留.BACKGROUND The immune rejection of the recipient is the main factor affecting the therapeutic effect of hepatocyte transplantation,and ultraviolet can cause immunosuppression.Appropriate intensity of ultraviolet irradiation can not only reduce the immunogenicity of hepatocytes,but also avoid excessive damage to hepatocytes caused by ultraviolet irradiation,so as to better preserve the stability of hepatocytes and cell synthesis function.AIM To investigate the effect of ultraviolet radiation on the immunogenicity and biological activity of primary adult human hepatocytes.METHODS Hepatocytes were isolated from benign adult liver tissues by collagenase perfusion and divided into a control group(0 J/m^2)and four experimental groups with different UV irradiation intensities(200,350,550,and 750 J/m^2).Trypan blue and CCK-8 were used to detect the cell viability.Mitochondrial membrane potential changes were detected with JC-1 dye.The proliferation of recipient T cells was determined by mixed lymphocyte hepatocyte culture(MLHC).The levels of albumin and lactate dehydrogenase in culture supernatant were tested.RESULTS The viable rate of newly isolated hepatocytes was more than 90%.CCK-8 detection revealed that the viability of hepatocytes in the 200 J/m^2 group was the highest,which had no significant difference compared with that of the control group,but was significantly higher than that of other experimental groups.In the presence of JC-1 dye,the hepatocytes in the control group and the 200 J/m^2 group mainly exhibited red fluorescence,and brown(350 J/m^2),yellow green(550 J/m^2),and green(750 J/m^2)changes were noted with the increase of irradiation intensity.The OD value of the 200 J/m^2 group was the highest,which had no significant difference compared with that of the control group,indicating that the membrane potential of hepatocytes was stable and the cell activity was the best;with the increase of irradiation intensity,the membrane potential of hepatocytes decreased significantly.The MLHC test showed that the 20

关 键 词：成人原代肝细胞 免疫原性 紫外线 

分 类 号：R575[医药卫生—消化系统]