腺病毒介导的白介素10基因转导对鼠胰岛β细胞的保护作用

Protective effect of adenovirus-mediated interleukin 10 gene transduction on bete cell of mice

目的观察腺病毒介导的白介素10(IL-10)基因转导对体外及体内胰岛β细胞的保护作用。方法用携带有IL-10基因的重组腺病毒载体感染RINm5F细胞、昆明小鼠及链脲佐菌素(STZ)诱导的1型糖尿病小鼠,ELISA检测IL-10基因表达、胰岛素水平,病理切片及免疫组化观察胰岛炎积分。结果 RINm5F细胞中检测到IL-10基因及蛋白的有效表达,含IL-10的重组腺病毒(Ad-IL-10)可抑制STZ致细胞凋亡作用。昆明小鼠腹腔注射Ad-IL-10后,再经STZ诱发为1型糖尿病,Ad-IL-10组胰岛炎积分明显降低。经STZ诱导的早期1型糖尿病小鼠,Ad-IL-10组与其他组相比平均血糖水平差别无统计学意义,小鼠胰岛炎症浸润程度与糖尿病对照组差别无统计学意义。结论腺病毒介导的IL-10可在胰岛RINm5F细胞有效表达,并且具有抗凋亡的能力。IL-10基因能够保护胰岛β细胞功能,促进胰岛素分泌,减少小鼠的胰岛炎和糖尿病的发生。IL-10基因对于已发生1型糖尿病早期的昆明小鼠无显著的治疗作用。

Objective To investigate the protective effect of interleukin 10（IL-10）gene transduction on beta cell in vitro and in vivo.Methods Adenovirus vector containing IL-10 gene was transfered into RINm5F cells.KM mice and KM mice with streptozotocin（STZ）-induced diabetic mellitus.The levels of IL-10 and insulin were detected by ELISA method,and insulitis score was investigated by immunohistochemistry.Results The mRNA and protein level of IL-10 were detected in RINm5F cells after gene tranduction.Ad-IL-10 transduction inhibited the STZ-induced apoptosis of pancreatic beta-cells.KM mice received intraperitoneal injection of Ad-IL-10 were then induced to diabetic mellitus by STZ.The insulitis score was significantly lower in Ad-IL-10 injected diabetic group than in non-injected diabetic group.KM mice were received intraperitoneal injection of Ad-IL-10 after STZ-induced type 1 diabetic mellitus was established.The average blood glucose level had no significant difference between Ad-IL-10 injected diabetic group and non-injected diabetic group.The infiltration of pancreatitis also had no significant difference between these two groups.Conclusions Ad-mediated IL-10 can effectively expressed in pancreatic RINm5F cells and exhibits anti-apoptotic ability.Adenovirus-mediated IL-10 expression can protect beta-cell,promote insulin secretion,and reduce the occurrence of insulitis and diabetes in mice.Adenovirus-mediated IL-10 expression does not show therapeutic effect on type 1 diabetic mellitus induced by STZ.