肝细胞生长因子对2型糖尿病小鼠创伤性口腔溃疡及外周血免疫细胞亚群的影响

Effect of HGF on wound healing and peripheral blood immune cell subsets in type 2 diabetic traumatic oral ulcer mice

目的:探讨肝细胞生长因子(hepatocyte growth factor,HGF)过表达对2型糖尿病小鼠口腔黏膜创伤性溃疡愈合及外周血免疫细胞亚群的影响。方法:野生型WT及HGF过表达转基因(HGF-Tg)小鼠各40只,随机分为普通饮食(ND)组及高脂饮食(HFD)组,HFD组小鼠通过高脂饮食16周成功建立糖尿病模型。各组再随机分为对照组及溃疡(Ulcer)组。溃疡组小鼠利用右侧颊黏膜切口制备口腔黏膜溃疡模型。于溃疡第5天测量各组小鼠体重和空腹血糖后处死小鼠,取溃疡组织行病理检查和溃疡指数评分;流式细胞法检测各组小鼠外周血中CD4^(+)T细胞、CD8^(+)T细胞、Ly6G^(+)中性粒细胞和F4/80^(+)巨噬细胞比例。结果:高脂饮食16周能显著升高小鼠体重和空腹血糖。溃疡第5天病理结果显示HGF-Tg-HFD-Ulcer组小鼠较WT-HFD-Ulcer组溃疡病灶间质纤维化更明显,炎症细胞密度减低,且溃疡指数评分更低。WT-ND-Ulcer组小鼠外周血中Ly6G^(+)中性粒细胞比例明显高于WT-ND组;HGF-Tg-ND-Ulcer组和HGF-Tg-ND组小鼠各种炎症细胞比例无明显差异。HGF-Tg-HFD-Ulcer组小鼠CD4^(+)、CD8^(+)T细胞占比明显低于WT-HFD-Ulcer组,且CD4^(+)/CD8^(+)T细胞比值更低。结论:HGF高表达可以减轻糖尿病小鼠溃疡组织的炎症反应,在促进口腔溃疡愈合的过程中起到积极作用。

Objective:Investigate the effect of hepatocyte growth factor(HGF)overexpression on wound healing and peripheral blood immune cell subsets in type 2 diabetic traumatic oral ulcer model mice.Methods:Forty WT mice and 40 HGF-TG mice were randomly divided into the normal diet(ND)group and the high fat diet(HFD)group,respectively.The HFD group mice successfully established the diabetes model through the high fat diet for 16 weeks.Mice in each group were randomly divided into a control group or a ulcer group.The model of mouth ulcer was built through an incision made in the right buccal membrane.The body weight and fasting blood glucose of mice in each group were measured on the 5 th day of mouse ulcer.The ulcer tissues were taken for pathological examination and ulcer index score examination.Meanwhile,the proportion of CD4^(+)T cells,CD8^(+)T cells,Ly6G^(+)neutrophils and F4/80+macrophages in peripheral blood of mice in each group were detected.Results:The weight and fasting blood glucose of mice could be significantly increased by high fat diet for 16 weeks.The pathological section showed that the ulcer involvement range and the inflammatory cells density of HGF-Tg-HFD-Ulcer group mice were smaller than that of WT-HFD-Ulcer group,meanwhile the score of ulcer index of HGF-Tg-HFD-Ulcer group mice were lower.The proportion of Ly6G^(+)neutrophils in peripheral blood of mice in WT-ND-Ulcer group was significantly higher than that in WT-ND group.The proportion of inflammatory cells between HGF-Tg-ND-Ulcer group and HGF-Tg-ND group has no significant difference.The proportion of CD4^(+)and CD8^(+)T cells in HGF-Tg-HFD-Ulcer group was significantly lower than that in WT-HFD-Ulcer group,and the ratio of CD4^(+)/CD8^(+)T cell was lower.Conclusion:HGF overexpression could reduce the ulcers and the inflammatory reaction of diabetic mice,and which might play a positive role in promoting the healing process of oral ulcer.