Exploring the effect and mechanism of osthole on diabetic foot ulcers in rabbits based on the AMPK/SIRT1 signaling pathway
WANG Jian-dong,FENG Bo
Department of Hand, Foot, and Ankle Surgery, The Third Affiliated Hospital of Inner Mongolia Medical University, Baotou014010, China
Abstract:
Objective To observe the therapeutic effect of osthole (OST) on diabetic foot ulcer (DFU) in rabbits and to explore its mechanism by intervening in the AMP-activated protein kinase (AMPK)/silent information regulator 1(SIRT1) pathway. Methods Seventy-two New Zealand rabbits were randomly divided into 6 groups (n=12 per group) using a random number table: sham operation group, DFU group, low dose OST(L-OST) group, high dose OST (H-OST) group, metformin group, and H-OST+AMPK inhibitor Compound C (H-OST+Compound C) group. Except for the sham operation group, DFU models were established in all other groups. Blood glucose meter was used to detect FBG. The software was used to analyze the wound healing rate. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), superoxide dismutase (SOD), malondialdehyde (MDA), insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF). HE staining was used to detect the wound granulation tissue morphology. Immunohistochemistry was used to detect the positive expression of CD31 protein. Moreover, Western blot was used to detect the expression of proteins related to the AMPK/SIRT1 pathway. Results In the DFU group, fibroblasts and neovascularization were relatively scarce, and inflammatory cell infiltration was significant. Compared with the sham operation group, the DFU group showed increased levels of FBG, TNF-α, IL-6, and MDA, as well as decreased levels of SOD, IGF-1, VEGF, positive expression rate of CD31, p-AMPK/AMPK, and SIRT1 (P <0.05). Compared with the DFU group, the L-OST, H-OST, and metformin groups exhibited improvements in the above indicators. In contrast, the H-OST+Compound C group showed opposite trends to those observed in the H-OST group. Conclusion OST exerts a significant therapeutic effect on DFU in rabbits, and its mechanism may involve the activation of the AMPK/SIRT1 pathway.
