The Study On The Therapeutic Effect Of Buyang Huanwu Decoction On Diabetes Wounds By Regulating AGEs-RAGE Signal Pathway

Objective:To investigate the therapeutic effects of Buyang Huanwu decoction on diabetic wounds,and to explore the mechanism of Buyang Huanwu decoction in treating diabetic wounds based on AGEs-RAGE signaling pathway.Methods:Low dose streptozotocin was injected intraperitoneally to cause diabetes in C57BL/6J mice(Streptozotocin,STZ).Skin perforations on the back’s two sides served as the foundation for the skin wound model.The mice were put into six groups at random:the normal control group(NC group),the diabetes model control group(DM group),the positive control group(metformin plus cilostazol,M+C group),the low dosage Buyang Huanwu decoction group(BYHWT-L group),the middle dose Buyang Huanwu decoction group(BYHWT-M group),and the high dose Buyang Huanwu decoction group(BYHWT-H group).15 mice in each group.24 hours after the establishment of the model,the BYHWT-L group,BYHWT-M group and BYHWT-H group were intragastrically perfused with the decoction of Buyang Huanwu decoction 9.26g/kg/d,18.53g/kg/d,37.06g/kg/d,respectively.and M+C group received 129 mg/kg/d of metformin hydrochloride tablet and 30.3 mg/kg/d of cilostazol tablet.and the NC group and DM group were perfused with distilled water of the same volume for 14 days.The overall health and wound-healing area of the mice in each group were assessed following modeling,and the skin wound healing rates of the mice in each group were noted at three days,five days,seven days,ten days,twelve days,and fourteen days.Hematoxylin-eosin staining and Masson trichrome staining were used to observe the histopathological alterations and collagen deposition region of the wound skin 15 days after modeling.Enzyme-linked immunosorbent assay(Elisa)was used to identify the expression of tumor necrosis factor-α(TNF-α)and interleukin-6,IL-6 inflammatory factors in the serum of mice in each group.In order to determine the positive expression rates of advanced glycation end products(AGEs),advanced glycation end products receptor(RAGE),vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1)and matrix metalloproteinase-2(MMP-2)in the wound skin of mice in each group,immunohistochemical staining was performed.the relative expression levels of RAGE,VCAM-1,ICAM-1,and MMP-2 mRNA in wound tissue of mice in each group were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR),and the protein expression levels of RAGE,VCAM-1,ICAM-1,and MMP-2 in wound tissue of mice in each group were detected by Western Blot(WB).The aforementioned experimental techniques were employed in this study to examine the therapeutic effects of Buyang Huanwu decoction on diabetic wounds as well as its mechanism of action.Results:1.The diabetic mouse model was successfully created by injecting STZ intraperitoneally,and mice whose blood glucose levels were within the normal range were chosen.With these mice,a back wound model for mice was created.Following intragastric delivery of mice to each group,the wounds in the DM group healed slowly,had a disorganized skin structure,and had less collagen fibers.Decoction of Buyang Huanwu can dramatically speed up wound healing,reduce wound size,and improve wound healing symptoms in diabetic mice.According to the results of H&E and Masson staining,Buyang Huanwu decoction can increase the collagen fibers in diabetic mice’s wound skin as well as the structure of their wounds.2.According to ELISA data,Buyang Huanwu decoction dramatically lowered the levels of TNF-α and IL-6 in diabetic wound mouse serum.Immunohistochemical staining results demonstrated that Buyang Huanwu decoction could significantly reduce the positive expression of AGEs,RAGE,ICAM-1,VCAM-1,and MMP-2 in the AGEs-RAGE signal pathway.RT-qPCR and Western Blot results demonstrated that Buyang Huanwu decoction could significantly reduce the mRNA and protein expression of RAGE,ICAM-1,VCAM-1,and MMP-2.Conclusion:In diabetic mice,Buyang Huanwu decoction can enhance the pathological alterations and the expression of inflammatory factors,boosting the healing of diabetic wounds.This suggests that Buyang Huanwu decoction has a certain encouraging effect on the treatment of diabetic wounds.Furthermore,Buyang Huanwu decoction can lower the expression of AGEs,RAGE,ICAM-1,VCAM-1,and MMP-2 molecules in the AGEs-RAGE signal pathway,suggesting that Buyang Huanwu decoction may treat diabetic wounds by affecting the AGEs-RAGE signal pathway.