摘要
目的探讨链脲佐菌素(STZ)注射诱导的1型糖尿病(T1DM)模型鼠胰岛细胞的损伤机制。方法STZ对清洁级雄性7周龄Balb/C小鼠胰岛细胞的直接细胞毒试验;应用小剂量STZ、多次腹腔注射方法,诱导1型糖尿病小鼠模型。然后,脱颈处死1型糖尿病模型鼠,无菌制备模型鼠脾淋巴细胞与无菌制备的预先小剂量STZ致敏的同系鼠正常胰岛细胞体外混合培养,以同系正常鼠脾淋巴细胞为对照,观察模型鼠脾淋巴细胞及STZ对胰岛细胞的影响,MTT法检测OD值,化学发光法检测各组胰岛细胞所分泌的胰岛素量。结果细胞毒试验显示以0.2 mg/ml STZ与正常Balb/C小鼠的胰岛细胞混合培养后,胰岛细胞损伤与对照组比较无明显变化(P>0.05),可用于体外胰岛细胞致敏;在脾淋巴细胞浓度为5×106个/ml时,混合培养72h和144 h的结果:无STZ致敏组、24hSTZ致敏组和48hSTZ致敏组与其各自对照组比较胰岛细胞的损伤及胰岛素的分泌量均无明显变化,而混合培养216h的结果:无STZ致敏组和24hSTZ致敏组与其各自对照组比较具有明显差异(P<0.05),48hSTZ致敏组与其对照组比较具有显著差异(P<0.01)。在脾淋巴细胞浓度分别为5×107/ml时,混合培养72 h的结果:无STZ致敏组和24 hSTZ致敏组与其各自对照组比较具有明显差异(P<0.05);48h STZ致敏组与其对照组比较具有显著差异(P<0.01),而混合培养144 h和216 h的结果:无STZ致敏组2、4 h STZ致敏组和48 hSTZ致敏组与其各自对照组比较具有显著差异(P<0.01)。结论T1DM模型鼠脾淋巴细胞对同系正常鼠的胰岛细胞具有一定的免疫损伤作用;糖尿病模型鼠脾淋巴细胞明显加强对低剂量STZ致敏的同系正常鼠胰岛细胞的免疫损伤作用。胰岛细胞的免疫损伤的程度与1型糖尿病模型鼠激活的脾淋巴细胞数量、低剂量STZ致敏同系鼠胰岛细胞的时间以及上述两者混合培养的时间密切相关。
Objective In vitro,explore STZ rats islet cell damage mechanism for type 1 diabetes,in the pathogenesis and treatment strategies provide the basis for in-depth study. Methods STZ＇ s direct cytotoxic effect on the islet cells of clean 7-week-old male Balb / C mices; Mices are induced model of type 1 diabetes by repeated intraperitoneal injection of small dose STZ. Then, type 1 diabetic mice were killed from cervical, its spleen lymphocytes of being taken out support with pancreatic islet cells of low-dose STZ sensitized normal mice in aseptic environment. To the department of normal mice with spleen cells as the control, observing the influence of spleen lymphocytes of model mouse and STZ on the islet cells. OD value of islet cells was detected by M3T, at the same time, insulin volume was detected by chemiluminescence. Results 0.2 mg/ml STZ has no apparent cytotoxic effect on islet cells; At splenocytes concentration of 5 × 10^6/ml,the results of mixed culture for 72 h and 144 h,without STZ sensitized group,24 h STZ sensitized group and 48 h STZ sensitized group compared with their respective control group show no significant change; The results of mixed culture for 216h,without STZ sensitized group and 24 h STZ sensitized group have a significant difference（ P 〈 0.05） ,48h STZ sensitized group have a very significant difference （ P 〈 0.01）. At splenocytes concentration of 5 × 10^7/ml, the results of mixed culture for 72 h,compared with their respective control group,without STZ sensitized group and 7Ah STZ sensitized group have a significant difference （ P 〈 0.05） ,48 h STZ sensitized group show a very significant difference （ P 〈 0.01） ;The results of mixed culture for 144 h and 216 h,without STZ sensitized group,24h STZ sensitized group and 48 h STZ sensitized group compared with their respective control group show a very significant difference （ P 〈 0.01 ）. Conclusion T1DM mouse model splenocytes have a certain immune injury effect on the islet cells of normal mice;T1DM mouse
出处
《中国实验诊断学》
2008年第9期1096-1099,共4页
Chinese Journal of Laboratory Diagnosis
