Intramuscular injection of interleukin-10 plasmid DNA prevented autoimmune diabetes in mice  

肌肉注射白介素-10质粒DNA预防小鼠自身免疫性糖尿病(英文)

作　　者：章振林[1] 沈水仙[1] 林波[2] 余路阳[2] 朱丽华[2] 王卫平[1] 罗飞宏[1] 郭礼和[2] 

机构地区：[1]复旦大学附属儿科医院内分泌和代谢科 [2]中国科学院上海生命科学院生物化学和细胞生物学研究所,上海200031

出　　处：《Acta Pharmacologica Sinica》2003年第8期25-30,110-111,共8页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China (№ 39993430).

摘　　要：AIM: To investigate the effect of plasmid coding intedeukin-10 （IL-10） DNA on the development of autoimmune diabetes induced by multiple low doses of streptozotocin （STZ） in mice. METHODS: Injection of STZ （40 mg/kg, ip） was given daily for five consecutive days. pcDNA3-IL-10 plasmid （IL-10-treated group） or pcDNA3-null plasmid （pcDNA3-null-treated group） （100 μg DNA once a day） were injected into skeletal muscles of mice on d 1 and d 14. Blood glucose concentration was measured. After mice were killed on d 28, serum IFN-γ level was measured by ELISA, and pancreatic IL-1β and TNF-α mRNA expression was detected by semi-quantitative reverse-transcription PCR （RT-PCR）. The number of CD⁴⁺ and CD⁸⁺ lymphocytes from spleen was detected using FACS. In addition, pancreatic histology was measured for determination of insulitis grades. RESULTS: Treatment with pcDNA3-IL-10 resulted in the retention and expression of the vector in skeletal muscle, associated with a considerable elevation in the plasma level of IL-10, which was not observed in pcDNA3-null-treated mice. In IL-10-treated diabetic mice induced by STZ, delay-type hypersensitivity responses were suppressed and the glucose level was greatly lower on d 14, 21, and 28 than pcDNA3-null-treated group （P【0.05 or P【0.01）. On d 21 and 28 the incidence of diabetes was 33.3% and 40.0%, respectively, which was markedly lower than that of pcDNA3-null-treated group （P【0.05）. In IL-10-treated mice pancreatic IL-1β and TNF-α mRNA expression was depressed, and serum IFN-γ concentration and the number of spleen CD4⁺ or CD8⁺ lymphocytes were decreased on d 28. The insulitis grades of IL-10-treated mice were lower than that of pcDNA3-null-treated group （P【0.01）. CONCLUSION: Systemic administration of IL-10 plasmid DNA can alleviate insulitis of experimental autoimmune diabetes in mice and reduce incidence of diabetes.目的:研究编码白介素-10(IL-10)质粒DNA对由多次、低剂量链脲佐菌素(STZ)诱发的自身免疫性糖尿病小鼠的作用。方法:连续5天将STZ(每次40 mg/kg)腹腔注射入小鼠体内,于第1、14天将100 mg表达人IL-10-pcDNA3质粒(IL-10处理组)或pcDNA3质粒(对照组)注射入骨骼肌内。测定小鼠血糖水平。第28天处死小鼠,检测小鼠血清干扰素-γ(IFN-γ)水平、胰腺IL 1β和TNF-α mRNA表达,测定脾CD4^+和CD8^+淋巴细胞的数量,同时进行胰腺组织学检查。结果:IL-10质粒DNA注射后,骨骼肌有IL-10 mRNA的持久表达,血浆IL-10水平明显升高,而且对迟发性超敏反应具抑制作用。IL-10处理组,在14、21和28天小鼠血糖显著降低,在21、28天时糖尿病发病率分别为33.3％和40.0％,显著低于对照组(P<0.05);胰腺IL-1β和TNF-αmRNA表达、血清IFN-γ水平,以及脾CD4^+和CD8^+淋巴细胞数量均低于对照组,胰岛炎症程度也明显轻于对照组(P<0.01)。结论:IL-10基因治疗能减轻实验性自身免疫性糖尿病小鼠 的胰岛炎症,降低糖尿病发生的机率。

关 键 词：INTERLEUKIN-10 insulin-dependent diabetes mellitus gene therapy intramuscular injections SPLEEN tumor necrosis factor interferon type Ⅱ MICE 

分 类 号：R9[医药卫生—药学]