| ObjectiveTo investigate the expression and clinical significance of Peroxiredoxin-2（PRDX2）in wound granulation tissue before and after negative pressure wound treatment（NPWT）in diabetic foot ulcers,so as to provide a new theoretical basis for revealing the mechanism of NPWT in promoting wound healing of chronic lower limb ulcer.Methods From June 1,2020 to November 31,2021,fifty-two healthy volunteers were selected from the health management center of the Hospital（NC group）,48 patients with type 2diabetic foot ulcer who were hospitalized in Department of Endocrinology（DFU group）,48 patients with T2 DM without complications（DM group）,and 16 patients with non-diabetic chronic lower limb ulceration in NPWT who were hospitalized in Department of Burn（SUC group）.DFU group is divided into pre DFU group and post DFU group,and SUC group is divided into pre SUC group and post SUC group.The general data,fasting peripheral venous blood of all subjects and wound granulation tissues of patients in DFU group and SUC group before and after NPWT were collected.The expression of PRDX2 in serum was measured by enzyme linked immunosorbent assay（ELISA）;The relative expression of PRDX2 in wound granulation tissue before and after DFU group and SUC group was measured by Western blot,immunohistochemistry and fluorescence quantitative PCR;The oxidative stress markers in wound granulation tissue before and after DFU group were detected by relevant detection kits,including superoxide dismutase（SOD）activity,malondialdehyde（MDA）content and glutathione（GSH）content.The changes before and after treatment were calculated according to the index values measured before and after NPWT in DFU group and suc group（Δ Value）.Results statistical analysis was performed by SPSS 25.0software.All P values were double-sided test.P<0.05 was determined as statistically significant.Results1.The results of serum PRDX2 concentration measured by ELISA showed that there was no significant difference between DM group,pre DFU group and NC group（P>0.05）,and the pre SUC group decreased compared with NC group,and the difference was statistically significant（P<0.001）.There was no significant difference between the pre DFU group and the DM group（P>0.05）,and the pre SUC group was lower than the DFU group,and the difference was statistically significant（P=0.003）.Compared with the pre DFU group,the pre SUC group decreased and the post DFU group increased,the difference was statistically significant（P < 0.05）.There was no significant difference between the post SUC group and the pre SUC group（P=0.553）.In addition,the change of serum PRDX2 concentration in DFU group before and after NPWT was significantly different from that in SUC group before and after NPWT（P=0.015）.2.The expression of PRDX2 protein in wound granulation tissue of each group was detected by Western blot showed that there was no significant difference between pre DFU group and pre SUC group（P=0.059）.Compared with the pre DFU group,the post DFU group was increased,and the difference was statistically significant（P=0.016）;Compared with the pre SUC group,the post SUC group increased,and the difference was statistically significant（P=0.012）.In addition,there was no significant difference in the relative changes of PRDX2 protein in wound granulation tissue between DFU group and SUC group before and after NPWT（P=0.462）.3.The expression of PRDX2 m RNA in wound granulation tissue of each group was detected by fluorescence quantitative PCR showed that there was no significant difference between pre DFU group and pre SUC group（P=0.070）.Compared with the pre DFU group,the post DFU group increased,and the difference was statistically significant（P < 0.001）;Compared with the pre SUC group,the post SUC group increased,and the difference was statistically significant（P = 0.001）.There was no significant difference in the relative changes of PRDX2 m RNA in wound granulation tissue between DFU group and SUC group before and after NPWT（P=0.110）.4.The average optical density of PRDX2 in wound granulation tissue of each group was detected by Immunohistochemical showed that there was no significant difference between the pre DFU group and the pre SUC group（P=0.896）.Compared with the pre DFU group,the post DFU group increased,and the difference was statistically significant（P < 0.001）;Compared with the pre SUC group,the post SUC group increased,and the difference was statistically significant（P = 0.001）.There was no significant difference in the relative changes of the average optical density of PRDX2 in wound granulation tissue between DFU group and SUC group before and after NPWT（P=0.706）.5.The results of oxidative stress markers showed that compared with the pre DFU group,the activity of SOD and the content of GSH in the post DFU group increased,while the content of MDA decreased.The differences were statistically significant（SOD: P<0.001;GSH: P<0.001;MDA: P<0.001）.6.Pearson or Spearman correlation analysis showed that the concentration of PRDX2 in DFU group was positively correlated with SOD activity（r = 0.266,P = 0.031）,negatively correlated with MDA content（r =-0.298,P = 0.015）,but not with GSH content（r=0.184,P=0.139）;The protein expression of PRDX2 in wound granulation tissue of DFU group was positively correlated with SOD activity and GSH content（SOD: r=0.528,P=0.036;GSH: r=0.556,P=0.025）,and negatively correlated with MDA content（r =-0.797,P < 0.001）;In DFU group,ΔPRDX2 protein in wound granulation tissue was negatively correlated with Δ WBC,Δ NEUT and Δ CRP（ΔWBC:r=-0.598,P<0.001;Δ NEUT:r=-0.369,P=0.023;Δ CRP:r=-0.446,P=0.005）,but not with Δ PCT;In group SUC,ΔPRDX2 concentration was positively correlated with Δ CRP（r=0.582,P=0.018）,but not correlated with Δ WBC,Δ NEUT and Δ PCT.ΔPRDX2 protein in wound granulation tissue was negatively correlated with Δ CRP and Δ PCT（Δ CRP:r=-0.968,P<0.001;Δ PCT:r=-0.628,P=0.009）,but had no correlation with Δ WBC and Δ NEUT.There was a positive correlation between the changes of serum PRDX2 concentration,PRDX2 protein expression in wound granulation tissue before and after NPWT with the wound healing rate at 4weeks in DFU group（serum: r=0.338,P=0.019;wound granulation tissue: r=0.329,P=0.023）;However,there was no correlation between the changes of serum PRDX2 concentration,PRDX2 protein expression in wound granulation tissue before and after NPWT with the wound healing rate at 4 weeks in SUC group.Conclusions1.PRDX2 is up-regulated in the wound granulation tissue of chronic lower limb ulcer after NPWT.2.The high expression changes of PRDX2 protein in wound granulation tissue of DFU patients before and after NPWT is related to wound healing. |
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