小肠黏膜下层仿生双层敷料复合富血小板血浆促进难愈性皮肤溃疡的修复

Small intestine submucosa bilayer bionic wound dressing combined with platelet-rich plasma promotes the repair of refractory skin ulcer

目的探讨小肠黏膜下层(SIS)仿生双层敷料复合富血小板血浆(PRP)对大鼠难愈性皮肤创面修复的影响。方法 40只SD大鼠随机分为对照组(Control组)、SIS修复组(SIS组)、PRP修复组(PRP组)、SIS复合PRP修复组(SIS+PRP组),对大鼠背部直径3 cm的区域进行25 Gy的辐照后,于辐照区制作1 cm×1 cm全层皮肤缺损,SIS+PRP组创面予以滴加了0. 2 m L PRP的SIS仿生双层敷料覆盖后外贴手术贴巾,SIS组创面予以滴加了0. 2 m L PBS的SIS仿生双层敷料覆盖后外贴手术贴巾,PRP组创面予以滴加0. 2 m L PRP后外贴手术贴巾,对照组创面予以滴加0. 2 m L PBS后外贴手术贴巾。观察各组大鼠术后第3,7,11,15天的创面愈合情况。采用酶联免疫吸附实验检测术后15 d时大鼠难愈性皮肤创面的MDA含量及SOD活性,采用蛋白质免疫印迹检测Nrf2信号通路相关蛋白的表达情况。结果 SIS+PRP组在术后第3,7,11,15天的愈合率均优于其他组,差异具有统计学意义(P <0. 05)。术后15 d,SIS+PRP组皮肤创面的MDA含量(5. 58±0. 86)μmol/g显著低于对照组(13. 71±2. 04)μmol/g、SIS组(11. 26±1. 79)μmol/g、PRP组(10. 18±1. 73)μmol/g,差异具有极显著性统计学意义(P <0. 01); SIS+PRP组皮肤创面的SOD含量(17. 60±3. 89) U/m L显著高于对照组(5. 02±2. 05) U/m L、SIS组(7. 10±2. 41) U/m L、PRP组(11. 37±2. 96) U/m L,差异具有极显著性统计学意义(P <0. 01)。与对照组、SIS组和PRP组相比较,SIS+PRP组皮肤创面Nrf2的表达量显著升高(P <0. 01),p-AKT及p-GSK-3β的表达量显著升高(P <0. 01),而Fyn的表达量则下降(P <0. 05)。结论小肠黏膜下层复合富血小板血浆可以促进难愈性皮肤溃疡的修复,可能与Akt/GSK-3β/Fyn信号通路介导的Nrf2/ARE活化有关。

Objective To investigate the effect and mechanism of small intestine submucosa bilayer bionic wound dressing combined with platelet-rich plasma on promoting the refractory skin wound repair in rats.Methods Forty SD rats were randomly divided into control group,SIS repair group（SIS）,PRP repair group（PRP） and SIS combined with PRP repair group（SIS＋PRP）.After 25 Gy irradiation in the area with a 3 cm diameter on the back of the rats,a 1 cm × 1 cm full-thickness skin defect was made in the irradiated area.The wounds in the SIS＋PRP group were treated with 0.2 mL PRP drops on small intestine submucosa bilayer bionic wound dressing and then was covered with films.The SIS group wounds were treated with 0.2 mL PBS drops on small intestine submucosa bilayer bionic wound dressing and were attached with films.The wounds in the PRP group were treated with 0.2 mL PRP followed by films,and the wounds in the control group were treated with 0.2 mL PBS followed by films.The wound healing on the 3rd,7th,11th,and 15th day after operation were observed in each group of rats.Enzyme linked immunosorbent assay（ELISA） was used to detect the content of MDA and SOD activity in the refractory skin wounds on the 15th day after operation.The protein expression of Nrf2 signaling pathway was detected by Western blotting.Results The healing rates of SIS＋PRP group on the 3th,7th,11th and 15th day after operation were better than those of other groups,the differences were significant （ P 〈0.05） .On 15th day after surgery,MDA content in skin wounds of SIS＋PRP group （5.58±0.86）μmol/g was significantly lower than that of control group （13.71±2.04）μmol/g,SIS group （11.26±1.79）μmol/g,and PRP group （10.18±1.73）μmol/g,the differences were significant（ P 〈0.01）;SOD content in skin wounds of SIS＋PRP group （17.60±3.89）U/mL was significantly higher than that of control group （5.02±2.05）U/mL,SIS group （7.10±2.41）U/mL and PRP group （11.37±2.96）U/mL,the differences were statistically significant （ P 〈0.01） ;Compared with control group,SIS group and PRP group,the expression level of Nrf2 in the skin wound of SIS＋PRP group was significantly increased（ P 〈0.01）,and the expression levels of p-AKT and p-GSK-3β were significantly increased（ P 〈0.01）.However,the expression level of Fyn was decreased（ P 〈0.05）.Conclusion The combination of small intestine submucosa bilayer bionic wound dressing with platelet-rich plasma can promote the repair of refractory skin ulcers,which may be related to Nrf2/ARE activation mediated by Akt/GSK-3β/Fyn signaling pathway.