miRNA-15b-5p对糖尿病足大鼠溃疡愈合的影响及可能机制

Effect and possible mechanism of miRNA-15b-5p on ulcer healing in diabetic foot rats

目的探索miRNA-15b-5p对糖尿病足大鼠溃疡愈合的影响及可能机制。方法将54只SD大鼠随机分为对照组、miRNA-15b-5p agomir组、糖尿病组和miRNA-15b-5p antagomir组。RT-qPCR检测对照组和糖尿病组大鼠创面皮肤组织miRNA-15b-5p表达;对比各组大鼠创面形成后第1、4、7、10、14天创面愈合率;TUNEL检测各组大鼠溃疡创面组织细胞凋亡情况;ELISA检测各组大鼠血清IL-1β、TNF-α、IL-6和IL-18水平和ICAM-1、E-selectin水平;Western blot法检测各组大鼠创面皮肤组织中IKKβ、IKKα、p-IKKβ、p-IKKα、NF-κB的表达水平。结果miRNA-15b-5p在糖尿病大鼠创面组织中表达上调,miRNA-15b-5p过表达后,创面愈合速率减慢,细胞凋亡率升高,炎症因子IL-1β、TNF-α、IL-6、IL-18和黏附分子ICAM-1、E-selectin表达升高,大鼠创面组织中p-IKKβ、p-IKKα、NF-κB蛋白表达水平升高。抑制miRNA-15b-5p表达后,加快创面愈合速率,抑制细胞凋亡,减少炎症因子IL-1β、TNF-α、IL-6、IL-18和黏附分子ICAM-1、E-selectin表达,抑制大鼠创面组织中p-IKKβ、p-IKKα、NF-κB蛋白相对表达水平。结论抑制miRNA-15b-5p有助于减轻炎症反应,促进糖尿病足大鼠溃疡愈合,其作用机制可能与调控IKKβ/NF-κB信号通路有关。

Objective To explore the effect and possible mechanism of miRNA-15b-5p on ulcer healing in diabetic foot rats.Methods Fifty-four SD rats were randomly divided into control group,miRNA-15b-5p agomir group,diabetes group and miRNA-15b-5p antagomir group.The level of miRNA-15b-5p was detected by RT-qPCR in the traumatic skin tissues of control and diabetic control rats.The healing rate of trauma was compared between the groups on days 1,4,7,10 and 14 after trauma formation.TUNEL was used to detect apoptosis of cells in the ulcerated skin tissues of each group.ELISA was used to detect the levels of IL-1β,TNF-α,IL-6,IL-18,ICAM-1 and E-selectin in serum of rats in each group.Western blot was used to detect the expressions of IKKβ,IKKα,p-IKKβ,p-IKKαand NF-κB in the traumatized skin tissues of rats.Results The expression of miRNA-15b-5p was up-regulated in the wound tissue of diabetic rats.After the overexpression of miRNA-15b-5p,the wound healing rate was slowed down,the apoptosis rate was increased,and the expressions of IL-1β,TNF-α,IL-6,IL-18,ICAM-1 and E-selectin were increased.The expressions of p-IKKβ,p-IKKαand NF-κB in wound tissue of rats were increased.Inhibition of the expression of miRNA-15b-5p could accelerate the rate of wound healing,inhibit apoptosis of cells,and reduce the expressions of IL-1β,TNF-α,IL-6,IL-18,ICAM-1 and E-selectin.The relative expressions of p-IKKβ,p-IKKαand NF-κB protein in rat wound tissue were inhibited.Conclusion Inhibition of miRNA-15b-5p can reduce inflammation and promote wound healing in diabetic foot rats,and its mechanism may be related to regulation of IKKβ/NF-κB signaling pathway.