| Objective: To observe the therapeutic effect of Astragalus Injection on diabetic ulcer wound and explore its mechanism of promoting the healing of diabetic ulcer wound.Methods: 192 rats were divided into diabetic model group,epidermal growth factor group,astragalus injection small dose group,astragalus injection medium dose group and astragalus injection large dosage group by intraperitoneal injection of streptozotocin(STZ).Quantitative group.Eight weeks later,the open wound of full-thickness skin defect,diabetic ulcer(DU),was made on the back of rats with a diameter of 20 mm and deep myomembrane.Drug intervention was started on the 2nd day after wound modeling to observe wound healing rate and time.During wound healing process(7th day,14 th day,21 st day after modeling),skin tissues inside and around the wound were cut and HE staining was used to observe the changes of wound tissue morphology and structure.Immunohistochemistry was used to observe the changes of integrin beta-1 and integrin beta-1.The changes of K19 and PCNA levels were observed at the levels of Wnt1,Wnt3 a,beta-catenin,GSK3 beta,Cmyc and Cyclin D1 by Western Blot and Real Time-q PCR,respectively.From the perspective of cell signal transduction pathway and epidermal stem cells,the expression of Wnt1,Wnt3 a,beta-catenin,GSK3 beta,Cmyc and Cyclin D1 was centered around "epithelialization-Wnt/beta-Catenin signal transduction pathway-wound surface".To explore the mechanism of Astragalus Injection in promoting the healing of diabetic ulcer.Results: Ⅰ.Animal diabetes model: After STZ injection,the body weight and blood sugar of diabetic model group were significantly different from those of normal control group(P < 0.05).Ⅱ.Comparison of wound healing rate and wound healing time: The wound healing rate of DU group was significantly lower than that of normal group on the 3rd,7th,14 th and 21 th day after modeling(P < 0.01);the wound healing rate of high dose group of Astragalus injection was higher than that of DU group on the 14 th and 21 st day after modeling(P < 0.01).Compared with the normal wound group,the wound healing time of DU group was significantly longer(P < 0.01);among the drug intervention groups,the wound healing time of the high dose group of Astragalus injection was less than that of DU group(P < 0.05).Ⅲ.Immunohistochemical results of wound tissue showed that the expressions of K19,beta 1 integrin and PCNA in the DU group were significantly lower than those in the normal group on the 7th,14 th and 21 st day after wound modeling(P < 0.01).At these three time points,the expressions of K19,beta 1 integrin and PCNA in the wound tissue of the high dose group of Astragalus injection and the epidermal growth factor group were all lower than those of the normal group(P < 0.01).Compared with DU group(P < 0.01),the expression of K19,integrin beta 1 and PCNA in large and medium dose group of Astragalus injection was higher than that in small dose group of Astragalus injection(P < 0.05).Ⅳ.Western Blot results of wound tissue: On the 7th,14 th and 21 st day after wound modeling,the expression of beta-catenin,C-myc and Cyclin D1 protein in DU group was significantly lower than that in normal wound group(P < 0.01).The expression of GSK-3beta in DU group was lower than that in normal wound group(P < 0.01),and the expression of GSK-3beta in DU group was higher than that in normal wound group(P < 0.05).At these three time points,the expressions of beta-catenin,beta-catenin nucleus and C-myc in epidermal growth factor group,astragalus injection large and medium dose wound tissue were higher than those in DU group(P < 0.05).On the expression of GSK-3beta,the expressions of epidermal growth factor group and astragalus injection were higher on the 7th and 14 th day after wound modeling.The dose group was lower than DU group(P < 0.05).The expression of beta-catenin and beta-catenin in the high dose group was higher than that in the low dose group on the 7th,14 th and 21 st day after wound modeling(P < 0.01),and the expression of GSK-3beta in the high dose group was lower than that in the low dose group on the 14 th and 21 st day after wound modeling(P < 0.01).Ⅴ.Real Time-q PCR results of wound tissue showed that the expression of beta-Catenin m RNA in DU group was significantly lower than that in normal wound group(P < 0.01)and GSK3 beta expression was higher than that in normal wound group(P < 0.01)at 7,14 and 21 days after wound modeling.Compared with DU group,epidermal growth factor group,epidermal growth factor group and GSK3 beta expression in DU group were significantly higher than those in normal wound group(P < 0.01).The expression of beta-Catenin in wound tissue of high dose group of Astragalus injection was higher than that of DU group(P < 0.05),and the expression of GSK3 beta was lower than that of DU group(P < 0.05).On the 7th and 14 th day after wound modeling,the expression of beta-Catenin in high dose group of Astragalus injection was higher than that in low dose group of Astragalus injection(P < 0.05),while the expression of GSK3 beta was lower than that in small dose group of Astragalus injection.The dose group(P < 0.05).The expression of Wnt1 and Wnt3 a in wound tissue in DU group was significantly lower than that in normal wound group at three time points(P < 0.01).Compared with DU group,the expressions of Wnt1 and Wnt3 a in high dose group of Astragalus injection and epidermal growth factor group were higher than those in DU group(P < 0.05).The expression of Wnt3 a in high dose group was higher than that in low dose group(P < 0.01).Conclusion: Ⅰ.Astragalus injection can improve wound healing rate of diabetic ulcer,shorten wound healing time,effectively promote the healing of diabetic ulcer,and there is a dose-effect relationship.Ⅱ.Astragalus membranaceus injection may be related to the activation of Wnt/beta-catenin signaling pathway.By up-regulating the expression of Wnt1,Wnt3 a and beta-catenin,it can activate the transcription of downstream target genes C-myc and Cyclin D1,increase the expression of C-myc and Cyclin D1,and play a role in regulating Wnt/beta-catenin signaling pathway,thereby promoting the proliferation and differentiation of epidermal stem cells,thereby promoting the proliferation of wound granulation tissue and accelerating wound healing.The process of epithelialization promotes the healing of diabetic ulcer. |
