重组HSP65-6P277乳酸乳球菌疫苗对链脲佐菌素诱导的1型糖尿病小鼠的降糖作用  

Hypoglycemic effect of Lactococcus lactis vaccine containing HSP65-6P277 on streptozotocin-induced type 1 diabetic mice

作　　者：吴洁[1] 刘晓锐[1] 杨雪[1] 侯景[1] 徐茂磊[1] 申丽丽[1] 黄东成[1] 刘坤锋[1] 

机构地区：[1]中国药科大学生命科学与技术学院微基因药学实验室,南京210009

出　　处：《中国药科大学学报》2014年第1期106-110,共5页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.81172973,31270985);中央高校基本科研业务费研究生专项资助项目(No.JKY2011076);江苏高校优势学科建设工程资助项目~~

摘　　要：探讨表达HSP65-6P277蛋白的乳酸乳球菌活载体疫苗对链脲佐菌素(STZ)诱导的1型糖尿病小鼠的降糖作用及可能的作用机制。采用多次小剂量腹腔注射STZ建立1型糖尿病模型。将诱导型表达菌株Lactococcus lactis NZ9000pCYT∶HSP65-6P277和组成型表达菌株L.lactis NZ9000 pHJ∶HSP65-6P277及无关蛋白对照菌株L.lactis NZ9000 pCYT∶Nuc灌胃1型糖尿病小鼠(2×109CFU,200μL),每天灌胃1次,连续7 d,之后每周灌胃1次,持续16周。每月从小鼠眼眶静脉丛取血1次,血糖检测仪检测STZ造模小鼠血糖水平并称体重。4个月后将小鼠处死,取小鼠脾脏做脾细胞增殖实验,采用ELISA试剂盒分别检测脾细胞上清液中的IL-10和IFN-γ细胞因子水平以及血清IgG抗体水平。结果显示,与对照组相比,pCYT∶HSP65-6P277组和pHJ∶HSP65-6P277组可以一定程度上降低STZ造模小鼠血糖水平(P<0.05),并能维持其正常体重稳定,降低脾细胞针对HSP65和P277的增殖(P<0.05),脾细胞上清液中IFN-γ细胞因子水平明显降低(P<0.05),血清IgG抗体水平无明显差异(P>0.05)。To study the hypoglycemic effect and mechanisms of vaccine HSP65-6P277 protein on type 1 diabetic mice induced by streptozotocin（STZ）, Lactococcus lactis live vector was used to express the vaccine protein. The mouse model of type 1 diabetes mellitus were established by intraperitoneal injections of multiple low dose STZ（MLD-STZ）and divided into three groups. The inducible expression strain L. lactis NZ9000 pCYT ∶HSP65-6P277, constitutive expression strain L. lactis NZ9000 pHJ ∶HSP65-6P277 and control strains L. lactis NZ9000 pCYT ∶Nuc were respectively immunized to diabetic mice orally once daily（2×109 CFU, 200 μL）in the first week and once every week in the following weeks lasting for 16 weeks. Fasting blood glucose levels were measured once a month on blood samples taken from orbital venous plexus. Four months later, the mice were sacrificed. Spleen cell proliferation experiment was performed. IL-10 and IFN-γ cytokine levels of spleen cells and serum IgG levels were assayed using ELISA kits. The results showed that pCYT ∶HSP65-6P277 group and pHJ ∶HSP65-6P277 group have low blood glucose（P 〈 0. 05）, and maintain normal body weight, compared with control group（pCYT ∶Nuc）. On the other hand, cytokine IFN-γ levels were significantly lower（P 〈 0. 05）in spleen cells supernatant and spleen cell proliferation was reduced against HSP65 and P277（P〈 0. 05）, but serum IgG levels were not significantly different（P〉 0. 05）.

关 键 词：乳酸乳球菌 疫苗 P277 黏膜免疫 1型糖尿病 

分 类 号：Q78[生物学—分子生物学] R965[医药卫生—药理学]