He Hao, Yang Yanli, Zhang Li
Department of Endocrinology, The Fourth Hospital of Changsha(Changsha Hospital of Hunan Normal University), Changsha 410000, China. *Corresponding author, E-mail:
Department of Endocrinology, The Fourth Hospital of Changsha(Changsha Hospital of Hunan Normal University), Changsha 410000, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2024 Dec;40(12):1083-1088.
Objective To investigate the role and possible mechanism of glycogen synthase kinase-3 beta (GSK-3β)/cAMP response element binding protein (CREB) signaling pathway in regulating macrophage pyroptosis in the pathogenesis and development of diabetic foot ulcer (DFU). Methods Thirty rats were randomly divided into control group, DFU group and GSK-3β inhibited group, with 10 rats in each group. Fasting blood glucose (FBG) was detected by dynamic blood glucose detector. The wound healing of each group was observed and recorded. The histopathologic changes of the wound were detected by HE staining. The level of wound fibrosis was detected by Masson staining. The protein levels of GSK-3β, CREB, gasdermin E (GSDME) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in wound tissue were detected by Western blotting. The co-expression of F4/80, GSDME and NLRP3 in wound tissue was detected by immunofluorescence staining. The serum levels of IL-1β and IL-18 were detected by ELISA. Results Compared with the control group, FBG in DFU group was increased. Compared with DFU group, FBG in GSK-3β inhibition group was decreased. The wound healing rate of rats in the inhibited GSK-3β group was higher than that in the DFU group from day 3 to day 14, and the difference was significant on day 14. Therefore, samples from day 14 were used in the follow-up experiment. Compared with the control group, the wound tissue of rats in DFU group was significantly damaged with collagen deposition defect, and the expressions of GSK-3β, CREB and apoptosis-related proteins GSDME and NLRP3 were increased, and the co-expressions of F4/80 and GSDME, F4/80 and NLRP3 were increased. Serum levels of IL-1β and IL-18 were increased. Compared with DFU group, most of the wound tissues of rats in GSK-3β group were healed. Collagen deposition at the fracture was increased. The expressions of GSK-3β, CREB and GSDME, NLRP3 were decreased. The expression levels of F4/80 and GSDME were reduced, along with a decrease in the co-expression of F4/80 and NLRP3. Additionally, there was a reduction in serum concentrations of IL-1β and IL-18. Conclusion GSK-3β/CREB signaling pathway and macrophage pyroptosis are significantly up-regulated in DFU rats. Inhibition of this pathway can promote DFU healing and down-regulate macrophage pyroptosis level.
目的 探讨糖原合酶激酶-3β（GSK-3β）/环磷酸腺苷反应元件结合蛋白（CREB）信号通路在糖尿病足溃疡（DFU）发病机制及发展过程中调节巨噬细胞焦亡的作用及可能机制。方法 将30只大鼠随机分为对照组、DFU组和GSK-3β抑制组，每组10只。采用动态血糖仪检测空腹血糖（FBG）。观察并记录各组伤口愈合情况。通过HE染色检测伤口组织的组织病理学变化。采用Masson染色检测伤口纤维化程度。通过蛋白质免疫印迹法检测伤口组织中GSK-3β、CREB、gasdermin E（GSDME）和核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）的蛋白水平。通过免疫荧光染色检测伤口组织中F4/80、GSDME和NLRP3的共表达情况。采用酶联免疫吸附测定法（ELISA）检测血清白细胞介素-1β（IL-1β）和白细胞介素-18（IL-18）水平。结果 与对照组相比，DFU组FBG升高。与DFU组相比，GSK-3β抑制组FBG降低。从第3天到第14天，GSK-3β抑制组大鼠的伤口愈合率高于DFU组，第14天差异有统计学意义。因此，后续实验采用第14天的样本。与对照组相比，DFU组大鼠伤口组织损伤明显，胶原沉积缺陷，GSK-
