1型糖尿病患者淋巴细胞对胰岛β细胞的毒性作用探讨

Study in vitro on lymphocytotoxic action of patients with type 1 diabetes mellitus to islet β cells

为探讨 1型糖尿病患者外周血淋巴细胞是否对胰岛 β细胞起破坏作用 ,随机选择新发 1型糖尿病患者 16例 ,分离其外周血淋巴细胞 ,与胎儿胰岛β细胞于体外混合培养 ,同时设单纯淋巴细胞及β细胞培养组、正常对照组。培养过程中观察细胞形态及其生长情况 ,培养后第 2 4、72小时分别留取培养液测胰岛素、IL- 2含量 ,电镜下观察培养细胞形态。结果显示 ,1型糖尿病患者血淋巴细胞与胰岛β细胞混合培养组胰岛细胞生长不良 ,IL- 2产生增多 ,胰岛素分泌减少。第 2 4小时 IL- 2为 12 72 .4± 95 9.7pg/ ml,显著高于单纯淋巴细胞组及正常对照组 (P<0 .0 5 ) ,第 72小时为 892 .9± 917.2 pg/ ml,高于单纯淋巴细胞组 (P<0 .0 5 ) ;第 2 4小时胰岛素为 5 0 8.4±185 .0μU/ m l,低于单纯胰岛β细胞组及正常对照组 (P<0 .0 5 ) ,第 72小时为 483.2± 199.1μU / ml,低于正常对照组 (P<0 .0 5 )。电镜观察见混合培养组 β细胞有变性和破坏。表明 1型糖尿病患者外周血致敏淋巴细胞对胰岛β细胞有破坏作用 。

To investigate the harmful role of activated lymphocytes in peripheral blood of patients with newly onset type 1 diabetes mellitus on islet β cells.Infant islet cells were separated and incubated in different wells . The lymphocytes separated from 14 normal subjects and 16 in patients with type 1 diabetes mellitus mixed and co cultured with islet cells. Simple lymphocyte or islet cell group was control group. After 24 and 72 hours mixed culture, the supernate was collected for IL 2 and insulin determination. The cultured cells were examined with microscope and electron microscope. The islet cells cocultured with lymphocytes of patients grew defectively compared with controls. The content of IL 2 in supernate increased but insulin decreased. At the 24th hour incubation of mixed culture group,IL 2 concentration in culture was 1272.4±959.7 pg/ml, higher than normal subjects and simple lymphocyte group(P<0.05). At 72nd hour it was 892.9±917.2pg/ml in mixed culture group, higher than in simple lymphocyte group(P<0.05). Insulin concentration of mixed culture group at the 24th hour was 508.4±185.0 μU/ml, significantly lower than normal subjects and simple islet cell groups. It was 483.2±199.1μU/ml at the 72nd hour, significantly lower than that in normal subjects(P<0.05). The electron micrograph showed β cell's destruction in mixed culture group. This indicate that the activated lymphocytes from peripheral blood of type 1 diabetes mellitus patients can lead to damage on β cells, and influence the functions and morphology of islet β cells .