Study On The Mechanism Of Guhan Yuyang Gel In Treating Diabetic Ulcer Based On Notch Pathwa

Objective: To observe the efficacy of "Guhan Healing Gel" on wound healing of skin ulcers in diabetic rats,and to explore the mechanism of regulating Notch signaling pathway on diabetic ulcer healing.Methods: Sixty-five healthy male SD rats were randomly divided into two groups,15 in the normal control group and 50 in the diabetes model group.Diabetic model rats were treated with1% STZ intraperitoneal injection at 55mg/kg to induce diabetic changes,and normal group intraperitoneally injected with equal volume of sodium citrate buffer.After 1 week of adaptive feeding,rats in the successful modeling and normal control group were selected to make an open skin ulcer model by surgical method.Diabetic ulcer rats were randomly divided into 3 groups of15 rats,namely diabetic ulcer(DU)model group,Guhan healing gel group and rubber muscle ointment group.The normal control group and the DU model group were covered with sterile gauze,and the Guhan healing gel group and the rubber muscle cream group were respectively replaced with gauze applied to the drug.The wound healing of rats was observed on the 3rd,7th and 14 th days after molding,and the serum and granulation tissue of the ulcer surface were taken and sent for testing after the rats were sacrificed.The morphological changes of granulation in wound wounds in each group were observed by HE staining,the expression of IL-6 and TNF-αin serum was measured by ELISA,the expression of Notch1 and Hes1 m RNA was detected by Real Time-q PCR and the expression of Notch1 and Hes1 proteins in Western Blot was detected.Results:(1)On the 3rd,7th and 14 th days after dressing change,the wound healing rate of rats at the same time point in each group was lower(P<0.05)compared with the normal control group,and the wound healing rate of Guhan healer gel group and rubber muscle cream group was significantly increased(P<0.05)compared with the DU model group,and the wound healing rate of Guhan healing gel group was better than that of rubber muscle cream group(P<0.05).(2)The HE results showed that compared with the normal control group,the wound suppuration was more obvious in the diabetes model group,there was inflammatory exudate,and the wound shrinking was slow.The granulation tissue maturation rate of wound granulation tissue in the Guhan Yue group was faster than that in the DU model group and the rubber muscle cream group,and the effect was the best in each group.(3)ELISA results: With the passage of dressing change time,the expression of IL-6 and TNF-α in each group of rats first increased and then decreased.The contents of IL-6 and TNF-α in the DU model group were significantly higher than those in the normal control group(P<0.01).Compared with the DU model group,the IL-6 and TNF-αcontents of Guhan Healing Gel and Rubber Muscle Cream were reduced(P<0.01),and the IL-6and TNF-α contents of Gu Han Healing Gel were lower than those of Rubber Muscle Cream(P<0.01).(4)Real Time-q PCR results: Compared with the normal control group,the expression of Notch1 m RNA in the DU model group,Guhan Yue group and rubber muscle-growing ointment group was significantly increased(P<0.05);compared with the DU model group,the expression of Notch1 m RNA in the rubber muscle cream group and Guhan Yue gel group was significantly reduced(P<0.05);and the Notch1 of Guhan Yue Gel was significantly reduced The expression of m RNA was lower than that in the rubber muscle cream group(P<0.05),compared with the normal control group,the expression of Hes1 m RNA in the DU model group,Guhan Yue group and rubber muscle-growing ointment group was significantly increased(P<0.05),compared with the DU model group,the expression of Hes1 m RNA in the rubber muscle cream group and the Guhan Yue gel group was significantly reduced,(P<0.05),and the Hes1 of Gu Han Yue gel on the 3rd and 14 th days The expression of m RNA was lower than that in the rubber muscle cream group(P<0.05),and the difference between the Guhan healing gel group and the rubber muscle cream group on day 7 was not statistically significant.(5)The Western Blot test results showed that compared with the normal control group,the DU model group,The expression of Notch1 and Hes1 proteins in the Guhan Healing Group and the Rubber Healing Paste Group was significantly increased(P<0.05),compared with the DU model group,the expression of Notch1 and Hes1 Proteins in the Rubber Healing Muscle Paste Group and the Gu Han Healing Gel Group was significantly lower(P<0.05),and the expression of Notch1 and Hes1 Proteins in the Guhan Healing Gel Group was lower than that in the Rubber Healing Muscle Cream Group(P<0.05).Conclusion: Guhan ulcer gel can downregulate the expression of IL-6 and TNF-αinflammatory factors,inhibit the inflammatory response of wounds,effectively improve the wound healing rate and shorten the wound healing time in diabetic ulcer rats.Its therapeutic mechanism may be related to the Notch signaling pathway,which inhibits the inflammatory microenvironment of ulcers,promotes the growth of epidermal stem cells,and accelerates wound epithelialization by downregulating the expression of Notch1/Hes1,thereby promoting diabetic ulcer healing.