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The Effect of Qilong Healing Ointment on Wound Repair in Diabetic Foot Rat Model Based on SIRT1-Mediated Autophagy |
CAI Xinlun, ZHU Yan, LI Jun, et al |
Kunming Hospital of Traditional Chinese Medicine, Yunnan Kunming 650599, China |
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Abstract Objective: To explore the effect and molecular mechanism of Qilong Healing Ointment on wound repair in diabetic foot rat model based on SIRT1 signaling molecule and autophagy. Methods: Male SD rats were randomly divided into control group, model group, positive drug metformin group, Qilong healing ointment group, and Qilong healing ointment + SIRT1 inhibitor EX527 group, with 8 rats in each group. A rat model of diabetic foot was constructed by feeding with high sugar and high fat, intraperitoneal injection of streptozotocin chain, and excision of dorsum skin of the foot. Continuous glucose meter to detect fasting blood glucose (FBG) in rats. The wound healing rate of rats was observed and calculated. HE staining was used to detect pathological changes in wound tissues. The levels of IL-6, TNF-α, IL-1β in serum were detected by ELISA. The expression of SIRT1, Beclin1, p62, LC3 mRNAs in wound tissue were detected by RT-qPCR. The expression of SIRT1, Beclin1, p62, LC3Ⅱ/Ⅰ proteins in wound tissue was detected by Western blot. An immunohistochemical staining assay was used to detect the expression of SIRT1 protein in wound tissues. Results: Compared with the control group, the level of FBG in the model group increased (P<0.05), the wound healing rate decreased (P<0.05), a large number of inflammatory cells were infiltrated in the wound tissue and the neovascularization and fibroblasts significantly reduced, the levels of IL-6, TNF-α, IL-1β in serum increased (P<0.05), the expression of SIRT1, Beclin1, LC3Ⅱ/Ⅰ decreased in wound tissue and p62 increased (P<0.05). Compared with the model group, the level of FBG in the positive drug group and the Qilong healing ointment group decreased (P<0.05), the wound healing rate increased (P<0.05), the infiltration of inflammatory cells in the wound tissue decreased and the number of neovasculars and fibroblasts increased, the levels of IL-6, TNF-α, IL-1β in serum decreased (P<0.05), the expression of SIRT1, Beclin1, LC3Ⅱ/Ⅰ increased in wound tissue and p62 decreased (P<0.05). EX527 significantly inhibits the effect of Qilong Healing Ointment (P<0.05). Conclusion: Qilong Healing Ointment can promote autophagy and inhibit inflammatory response to alleviate the damage of wound tissue in rats with diabetic foot, and its mechanism may be related to the activation of SIRT1 signaling.
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