腺病毒介导的鼠IL-10基因对非肥胖型糖尿病小鼠胰岛β细胞的保护作用

Protective effect of adenovirus vector mediated interleukin 10 gene on islet beta cells of NOD mice

目的探讨腺病毒介导的鼠IL-10(Ad-rIL-10)基因对非肥胖型糖尿病(NOD)小鼠1型糖尿病早期胰岛β细胞的保护作用。方法随机将46只NOD小鼠分为4组:生理盐水对照组10只、糖尿病对照组12只、空载体对照组12只、Ad-rIL-10组12只。除生理盐水对照组外,其他组腹腔注射环磷酰胺诱导糖尿病早期模型。成模后,Ad-rIL-10组立即给予腹腔注射含Ad-rIL-10的重组腺病毒液0.1 mL、空载体对照组给予含Ad-eGFP的重组腺病毒液0.1 mL,其他组给予等量生理盐水。每周检测小鼠体质量、血糖。3周后处死小鼠,留取小鼠血清标本,采用ELISA法测定血清C肽、IFN-γ、IL-4及IL-10水平;制作胰腺标本,免疫组化法观察小鼠胰腺炎症浸润程度和rIL-10局部表达情况。结果与生理盐水对照组相比,成模后其他组血糖水平、体质量变化有明显差异(P均<0.05),血清IFN-γ水平明显增高(P均<0.05),而IL-4、IL-10和C肽水平明显降低(P均<0.05)。糖尿病对照组、Ad-rIL-10组、空载体对照组血糖水平、体质量、胰岛炎症浸润程度及血清学指标变化不明显。结论 Ad-rIL-10基因对NOD小鼠1型糖尿病早期胰岛炎症无明显减轻作用,对残余胰岛β细胞无明显保护作用。

Objective To explore the protective effect of adenovirus vector mediated murine interleukin 10 （Ad-rIL- 10） gene on islet beta cells at the early stage of type 1 diabetes in NOD mice. Methods Forty-six NOD mice were randomly divided into four groups：saline control group （ten mice） , diabetes control group（ twelve mice） , mock-vehicle control group（twelve mice） and Ad-rIL-lO group（twelve mice）. Except the saline control group, the rest mouse were received in- traperitoneal injection of cyclophosphamide in order to induce type 1 diabetes mellitus. After that, Ad-rIL-10 group were immediately received intraperitoneal injection of ad-rIL-10 gene at the dose of 0.1 mL, mock-vehicle control group were immediately received intraperitoneal injection of ad-eGFP at the dose of 0.1 mL, and the rest were given equal saline. Body weight and blood glucose of all mice were detected weekly. Three weeks later, all mice were executed, and the serum IL- 10, IL-4, INF-γ and C-peptide were detected by enzyme linked immunosorbent assay. Their pancreas were sectioned to observe the degree of insulitis and the local expression of IL-10 gene detected by immunohistochemistry. Results Compared with the saline control group, average blood glucose level and body weight of the other group had significant difference（ all P 〈0.05） , and serum IFN-γ level increased markedly（ all P 〈 0.05 ） , while IL-4, IL-10 and C-peptide level decreased significantly（P 〈 0.05 ）; IL-10 gene had a higher expression in Ad-dL-10 group than that in the other groups （all P 〈 0.05） ; Among diabetes control group, mock-vehicle control group, Ad-rIL-10 group, the change of blood glucose level, body weight, islet inflammatory infiltration and the serum index were not obvious. Conclusions The administration of Ad-rlL-10 gene dosen＇t significantly reduce the degree of insulitis, and the early protection on islet β cells of type 1 diabetes mellitus in NOD mice is not obvious.