Study On The Antioxidant Effect And Mechanism Of MEBO On Diabetic Skin Ulcer In Mice

Purpose:Diabetic skin ulcer is a well-recognized thorny problem in clinically refractory wounds.There are many comprehensive treatment methods in clinical practice,but the traditional Chinese medicine has a lack of treatment methods,especially the basic research on healing mechanism and antioxidant mechanism.This experimental study was conducted to observe the effect of MEBO on the wound healing and pathological changes in diabetic skin ulcer model mice,and further evaluate the intervention effect of MEBO on the wound healing mechanism of diabetic skin ulcer mice.By detecting the levels of SOD and GSH-px in serum and the expression of Nrf2,NQO1 and HO-1 in muscle tissue of skin lesions,the mechanism of MEBO on the anti-oxidation and oxidative stress of diabetic skin ulcer mice was investigated.Clinical application of MEBO in the treatment of diabetic skin ulcers provides more scientific experimental data.Material and method:Forty male SPF male Kunming mice,30 mice were fasted for 24 hours the day before the experiment,and the next day,streptozotocin(STZ)was used for intraperitoneal injection to induce a diabetic mouse model.After 72 hours of STZ injection The blood glucose was randomly detected 2 times or more,and greater than or equal to 16.7mmol/L to confirm the successful replication of the diabetic mouse model.The mice were then given a hair removal on the back,and the skin was made a "well" shaped incision on the back with a scalpel.The incision was about 1 cm long and was deep enough to expose the muscle layer.At the same time,the successfully replicated diabetic mouse model was randomly divided into 3 groups,10 in each group: model control group,MEBO group(experiment group),rehabilitation new fluid control group(positive control group),and other small health groups.The rats served as a blank control group.The positive control group and the experimental group were given rehabilitation fluid and MEBO respectively for topical drug dressing intervention in the wound,once a day for 7 days.After 1 week,the wound healing was observed continuously and recorded in detail.Whole blood was collected from the posterior venous plexus of the mice,and the levels of SOD and GSH-PX antioxidant enzymes in the serum were detected.The pathological changes of muscle tissue wereobserved by HE staining.The muscle tissue of the site is divided into two pieces,which are directly frozen and used for the detection of Nrf2,NQO1 and HO-1 expression levels by western-blot method;the other is fixed in 4% paraformaldehyde solution for IHC detection of Nrf2,NQO1,HO-1 expression levels.Results:1.Wound healing: In the MEBO group,the wound healing rate of the Kangfuxin group was significantly better than that of the model group(p<0.01),and the healing rate was faster.The MEBO group and the rehabilitation group had significant differences(p<0.01),which were statistically significant.2.HE staining observation: After 1 week of modeling,the pathology showed that there were significant differences between the four groups under the microscope;3.Colorimetric detection: The levels of SOD and GSH-px antioxidant enzymes in the serum of MEBO group and Huoxinxin group were significantly different from those of the model control group(p<0.01).MEBO group The levels of SOD and GSH-px antioxidant enzymes in the model group were significantly different from those in the blank control group(p<0.01),which was statistically significant.4.WB method and IHC method detection: The expression levels of Nrf2,NQO1 and HO-1in muscle tissue of mice in MEBO group and Kangxinxin group were significantly different from those in model control group(p<0.01).The expression levels of Nrf2,NQO1 and HO-1in the muscle tissue of the diabetic rats with burn ointment group and Kangfuxin group were significantly different from those of the control group(p<0.01),which was statistically significant.;Conclusion:1.The results of basic experimental research prove to some extent that MEBO improves the healing rate of diabetic skin ulcers and accelerates the healing rate of wounds.2.The expression of SOD and GSH-px in serum of mice with diabetic skin ulcer was significantly increased by topical MEBO.The expression of Nrf2,NQO1 and HO-1 protein in mouse skin lesions was significantly increased by WB method and IHC method.It fully revealed the antioxidant effect of MEBO on mice with diabetic skin ulcer and how Nrf2 interferes with the regulation of NQO1 and HO-1 protein expression and regulation.