| PurposesThis paper aimed to investigate the significance of miR-296-5p in diabetic chronic wounds healing.The study on the relationship between the effect of miR-296-5p on wound healing and Notch signaling pathway,would provide new ideas and clinical treatment for diabetic refractory wound.Methods(1)6 cases of non-diabetic patients with chronic wounds and 6 cases of diabetic lower extremity wound tissue were labeled as N group and D group.Real time fluorescent quantitative PCR determination for expression of miR-296-5p,PCR and Western-blot determination the level of Numbl-mRNA,Numbl protein,NICD and Hes-1.(2)36 diabetic rats were randomly divided into three groups.Through the local upregulation(labeled as B group)and downregulation(labeled as C group)of miR-296-5p,we compared them with control group(labeled A group).At 7th day and 14 th day after surgery,the expression of miR-296-5p,Numbl-mRNA,Numbl protein,NICD and Hes-1 were detected.We also calculated the rate of wound healing.Results(1)miR-296-5p in the N group the 2-delta CT value was(4.458±2.525)*10^-5,in the D group was(1.011±1.630)*10^-5,the expression of Numbl-mRNA was 0.359±0.122,diabetes group was 0.939 ±0.428.Western blot results: the Numbl protein were 0.41±0.17(N group)and 1.69±0.62(D group);NICD protein were 2.11±0.47(N group)and 0.86±0.36(D group);HES-1 transcription factor were 1.20±0.34(N group)and 0.54±0.16(D group),P<0.05.(2)After operation 7 days,the miR-296-5p in the three group of the 2-delta CT value were(1.299±0.464)*10^-5,(9.406±1.979)*10^-5,(0.368±0.324)*10^-5,P<0.05;14 days after surgery they were(1.224±0.665)*10^-5,(2.048±1.019)*10^-5,(0.872±0.319 *10^-5),no statistically significant difference.The expression of Numbl-mRNA on postoperative day 7 tissue samples were 0.819±0.188(A group),0.431±0.167(B group),2.310±0.749(C group),*P<0.05,14 days after surgery was 1.179±0.211(A group),0.962±0.253(B group),1.537±0.577(C group),no significant difference.Western blot results: the Numbl protein on the 7 postoperative day three group were 1.51±0.52(A group),0.71 ± 0.26(B group),2.80± 0.82(C group),and 14 days after surgery,followed by 1.34±0.35(A group),0.88±0.07(B group).1.85±0.22(C group);The level of NICD in 7 days after operation were 1.02±0.24(A group),1.84±0.37(B group),0.47±0.24(C group),and 14 days after surgery,followed by 0.85±0.15(A group),1.34±0.35(B group),0.59±0.13(C group);transcription factor HES-1,on the 7 postoperative day were 0.66 ± 0.11(A group),1.12 ± 0.31(B group),0.35 ± 0.14(C group),and 14 days after surgery,followed by 0.50 ± 0.13(A group),0.74 ± 0.13(B group),C(0.29 ± 0.11 group),P<0.05.The calculation results show that the wound area,the 7 day after operation in three groups were: 199±13.3(A group),189.5 ± 12.3(B group),206.3±8.38(C group),no significant difference;after 14 days: 39.5±11.4(A group),20.3±10(B group)51.3,± 14.9(C group),there were significant differences between groups A and B,mm2 unit.The number of healing days: 21.5±1.73 days(A group),18.5±1.29 days(B group)and 23.75±1.5 days(group C),there were significant differences between groups A and B.Conclusion(1)The expression of miR-296-5p decreased while the Notch signaling pathway was inhibited.(2)The increase of MiR-296-5p level can enhance the Notch signaling pathway and accelerate the healing of diabetes mellitus by lowering Numbl. |
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