Department of Pediatrics, Affiliated Hospital of Qingdao Medical College, Qingdao University, Qingdao, China.
Mol Cell Biochem. 2010 Nov;344(1-2):163-71. doi: 10.1007/s11010-010-0539-x. Epub 2010 Jul 25.
Interleukin-10 (IL-10) is a pleiotropic immunosuppressive and immunostimulatory cytokine. In autoimmune diabetes of the nonobese diabetic (NOD) mouse, IL-10 has exhibited paradoxical effects. Systemic IL-10 expression prevented or delayed diabetes onset in NOD mice while local expression of IL-10 did not. As antigen-presenting cells (APCs) play a central role in the generation of primary T cell responses, the direct role of this gene in pancreatic beta (β) cell is not clear. The effects of IL-10 on the protection of β cells in vitro were examined. In the present study, we examined the effects of adenovirus vector-mediated murine IL-10 (mIL-10) gene transfer to islet cell line RINm5F cells in vitro and to explore if IL-10 overexpression may prevent cytokine-mediated cytotoxicity. We had established the recombinant adenovirus vector containing mIL-10 genes (Ad-mIL-10) successfully. After infection of Ad-mIL-10, both mRNA and protein were expressed in RINm5F cells. Moreover, RINm5F cells secreted IL-10 protein into culture medium. Ad-mIL-10 prevented IL-1β-mediated nitric oxide production from β cells in vitro as well as the suppression of β cells function as determined by glucose-stimulated insulin production. Furthermore, Ad-mIL-10 gene transfer led to a profound reduction of Fas-expressing β cells and caspase-3 activity which were induced by IL-1β and the apoptotic rates of Ad-mIL-10 group were decreased. These findings show that IL-10 gene transfer to β cells may be beneficial in maintaining cells function, protecting islet cells from apoptosis-mediated by factors, which showed the potential therapy for type 1 diabetes mellitus.
白细胞介素-10(IL-10)是一种多效性的免疫抑制和免疫刺激细胞因子。在非肥胖型糖尿病(NOD)小鼠的自身免疫性糖尿病中,IL-10 表现出矛盾的作用。全身表达 IL-10 可预防或延迟 NOD 小鼠的糖尿病发病,而局部表达 IL-10 则没有。由于抗原呈递细胞(APC)在原发性 T 细胞反应的产生中发挥核心作用,该基因在胰岛β(β)细胞中的直接作用尚不清楚。本研究旨在探讨白细胞介素-10(mIL-10)基因转移对胰岛细胞系 RINm5F 细胞的体外保护作用,并探讨 IL-10 过表达是否可以预防细胞因子介导的细胞毒性。我们成功建立了含有 mIL-10 基因的重组腺病毒载体(Ad-mIL-10)。感染 Ad-mIL-10 后,RINm5F 细胞中表达了 mIL-10 的 mRNA 和蛋白。此外,RINm5F 细胞将 IL-10 蛋白分泌到培养基中。Ad-mIL-10 可预防 IL-1β介导的体外β细胞一氧化氮产生,并抑制β细胞功能,如葡萄糖刺激的胰岛素产生所测定的。此外,Ad-mIL-10 基因转移导致 Fas 表达的β细胞和 caspase-3 活性显著减少,这些都是由 IL-1β诱导的,Ad-mIL-10 组的凋亡率降低。这些发现表明,将 IL-10 基因转移到β细胞中可能有益于维持细胞功能,保护胰岛细胞免受细胞因子介导的凋亡,这为 1 型糖尿病的潜在治疗提供了依据。