| Objective: To explore the effect of ferulic acid（FA）on vascularization and tissue sugar content of diabetic wounds,to study whether it can promote the healing of diabetic wounds,and to provide experimental basis for clinical treatment.Methods:1.Experimental animal establishment: Thirty SPF SD female rats（8 weeks old,body weight 180.0-220.0g）were adaptively fed with common chow for 3 days and divided into 5 groups by random number table method: normal control group（NC）,diabetic control group（DC）,5% ferulic acid group（5% FA）,10% ferulic acid group（10% FA）and 15% ferulic acid group（15% FA）,6 per group.2.Type II diabetes mellitus modeling: The NC group was fed with ordinary feed,and the other 4 groups were fed with high-sugar and high-fat feed for 4 weeks.During the period,body weight and fasting tail vein blood glucose were measured weekly（During the experiment,the tail of the rat was stimulated every day to reduce the blood sugar fluctuation caused by the stress of the rat when the blood sugar was measured by the tail vein）.After 4 weeks,the rats were fasted for 20 hours but could drink water.The body weight and fasting blood glucose of the rats were measured.1.0% STZ（30 mg/kg）was intraperitoneally injected.The NC group was injected with the same amount of citric acid-sodium citrate buffer,group feeding was continued after STZ injection.Fasting blood glucose was measured on the 3rd and 7th day of STZ injection.Fasting blood glucose ≥ 16.7mmol/L,combined with obvious symptoms of polydipsia,polyphagia and polyuria,the diabetes modeling was considered successful.If the fasting blood glucose of the rat was lower than this standard after 7 days,the low-dose 1.0% STZ（20 mg/kg）was re-induced until the model was successfully established.A total of 24 diabetic rats were established.3.Diabetic trauma modeling: After the diabetes model was established,1.0%sodium pentobarbital 6.0 m L/kg was slowly injected intraperitoneally for anesthesia,the back hair of the rats was removed with a razor,a circle with a diameter of 20.0mm was marked with a marker pen,and the hair removal area was sterilized with type III medical iodophor,and cut out.The skin and subcutaneous tissue of the area were marked,and the bleeding was stopped by compression to make a rat diabetic wound model.The skin tissue was removed and stored in a refrigerator at-80°C,and the glucose content of the tissue was measured.4.Ferulic acid ointment preparation: Weigh 9.0 g of Vaseline on a tray balance and put it in a beaker,weigh 1.0 g of ferulic acid powder,and fully stir and mix in a45°C water bath to prepare 10.0% ferulic acid ointment.5% ferulic acid ointment and15% ferulic acid ointment were prepared in the same way.5.Wound management and observation: Vaseline was applied to the wounds of the NC group and DC group,5%,10%,and 15% ferulic acid ointment were applied to the wounds of the 5%FA group,10%FA group and 15%FA group respectively.The thickness of the smear is based on the color of the wound that is completely covered.The dressings were changed and the wounds were observed every day.Pictures were taken on the 0th day,7th day and 14 th day of the wound formation,and Image J image analysis software was used to calculate the wound area.On the 14 th day,after anesthesia,the wound tissue was excised for HE staining to observe histological changes,immunohistochemistry to detect the expression of CD34,and immunofluorescence to detect the expression of vascular endothelial growth factor（VEGF）,Image pro Plus（6.0）software for quantitative analysis.Glucose content detection kit was used to measure the glucose content of wound tissue by micro method.Results:1.Body weight and fasting blood sugar: At the end of adaptive feeding,there was no difference in body weight and fasting blood glucose among the groups（p>0.05）.After group feeding for 3 weeks,the body weight of DC group,5%FA group,10%FA group and 15%FA group was higher than that of NC group（P<0.01）,but there was no difference between groups（p>0.05）.The fasting blood glucose of rats fluctuated to varying degrees,but none of them met the diagnostic criteria of diabetes.After STZ injection,the weight gain rate of DC group and FA three groups slowed down or even decreased（p<0.01）,while the weight of NC group has been increasing steadily.Fasting blood glucose in DC group and FA three groups was significantly higher than that in NC group（p<0.01）,which met the diagnostic criteria of diabetes,and there was no difference in fasting blood glucose among the four groups（p>0.05）.After FA intervention,the fasting blood glucose in the FA three groups decreased（p<0.01）and was lower than that in the DC group（p<0.01）,but remained above the diagnostic level of diabetes,and the fasting blood glucose in the NC group was lower than the other four groups（p<0.01）.2.Wound healing: On the 7th day of wound formation,the wound healing rate in the NC group was higher than that in the other 4 groups（p<0.01）;the FA three groups was higher than the DC group（p<0.01）;the 5%FA group was the same as the 10%FA group（p>0.05）,but the 15%FA group was greater than the 5%FA group（p<0.05）.On the 14 th day,the wound healing rate of the NC group was higher than that of the other4 groups（p<0.01）;the FA three groups was higher than the DC group（p<0.01）,and the 10%FA group was the same as the 15%FA group（p>0.05）,but greater than 5%FA group（p<0.05）.3.Histological change: In the NC group,more collagen was seen in the wound,the wound was basically epithelialized,and there were few inflammatory cells.There were scattered blood vessels,and the vascular structure was relatively complete;in the DC group,more inflammatory cells were infiltrated,and the wound was still in a continuous and high-level inflammation stage;in the FA group,there was no obvious collagen formation,and the number of blood vessels was small,and the structure of the vessel wall was immature;more blood vessels were seen in the FA group,and more collagen tissue was seen in the FA group than in the DC group,and inflammatory cells and granulation tissue were found in the tissue.4.Vascular density: The NC group was higher than all other groups（p < 0.01）;the 5% FA,10% FA and 15% FA groups were higher than the DC group（p < 0.05）;there was no difference between the FA three groups（p > 0.05）.5.VEGF expression: The NC group was higher than the other 4 groups（p <0.01）;the 10% FA group and the 15% FA group were higher than the DC group（p <0.05）;there was no difference between the 5% FA group and the DC group（p > 0.05）;there was no difference between the FA three groups（p>0.05）.6 Tissue glucose content: On the day of wound formation,the glucose levels in the four diabetes model groups were the same（p>0.05）,but significantly higher than those in the NC group（p<0.01）.After 14 days of FA intervention,the 5%FA group,10%FA group and 15%FA group were lower than the DC group（p<0.05）;the 15%FA group was lower than the 10%FA group（p<0.01）;the 10%FA group was lower than the 5%FA group（p<0.01）;the glucose level in the NC group was lower than that in the other 4 groups（p<0.01）.Conclusion: Ferulic acid accelerates diabetic wound healing by reducing the glucose content of the wound tissue and promoting diabetic wound vascularization. |
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