Wang L, Guo F, Min D H, Liao X C, Yu S Q, Long X X, Ding X, Guo G H
Graduate School of Nanchang University, Nanchang 330006, China.
Burn Center of the First Affiliated Hospital of Nanchang University, Nanchang 330006, China.
Zhonghua Shao Shang Za Zhi. 2019 Jan 20;35(1):18-24. doi: 10.3760/cma.j.issn.1009-2587.2019.01.005.
To compare the tissue morphology and gene expressions of inflammatory and repair-related factors in chronic refractory wound tissue including pressure ulcers and diabetic feet. During August 2016 to September 2017, 10 samples of prepuce were collected after circumcision of 10 urological patients [all male, aged (38±4) years old] admitted in the First Affiliated Hospital of Nanchang University and included in normal skin group, samples of tissue around the edge of wounds with blood supply were collected from 9 heat or electric burn patients [6 male patients, 3 female patients, aged (51±8) years old], 13 pressure ulcer patients [9 male patients, 4 female patients, aged (51±14) years old] and 10 diabetic foot patients [8 male patients, 2 female patients, aged (61±10) years old] during the operations. The samples were divided into burn wound group (9 samples), pressure ulcer group (13 samples), and diabetic foot group (10 samples). Ten slices were taken from pressure ulcer group and diabetic foot group respectively, and 5 slices in each group were used to observe the tissue morphology and expressions of Ki67 and CD31 of wounds respectively with immunofluorescence method. Ten samples from normal skin group, 9 samples from burn wound group, 13 samples from pressure ulcer group, and 10 samples from diabetic foot group were collected for analysis of mRNA expressions of vascular endothelial growth factor 192 (VEGF192), transforming growth factor β (TGF-β), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) , interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α) by real time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with Mann-Whitney test and Kruskal-Wallis rank-sum test. (1) The expression level of Ki67 in diabetic foot group (390±100) was higher than that of pressure ulcer group (182±14, =-2.611, <0.01). (2) Although there were a large number of vascular endothelial cells (CD31 positive cells) in wounds of diabetic foot group, their distribution was disordered and failed to form intact lumen. There were less vascular endothelial cells in wounds of pressure ulcer group than those of diabetic foot group, but the complete lumen was formed. (3) The mRNA expression levels of VEGF192 in wounds of burn wound group, pressure ulcer group, and diabetic foot group were significantly lower than the level in normal skin group (=13.72, 30.50, 15.20, <0.05 or <0.01), and the level was the lowest in pressure ulcer group. The mRNA expression level of VEGF192 in wounds of pressure ulcer group was significantly lower than that of diabetic foot group (=15.30, <0.01). Compared with that of normal skin group, the mRNA expression level of TGF-β in wounds of burn wound group showed no significant difference (=-9.50, >0.05), while the mRNA expression levels of TGF-β in wounds of pressure ulcer group and diabetic foot group were significantly decreased (=18.04, 14.50, <0.01). The mRNA expression level of TGF-β in wounds of pressure ulcer group was similar to that of diabetic foot group (=3.54, >0.05). (4) Compared with those of normal skin group, the mRNA expression levels of VCAM-1 in wounds of burn wound group and pressure ulcer group were significantly increased (=-22.50, -11.50, <0.05 or <0.01), and there was no significant difference in the mRNA expression level of VCAM-1 in wounds of diabetic foot group (=10.00, >0.05); the mRNA expression level of ICAM-1 in wounds of burn wound group showed no significant difference (=-9.50, >0.05), and the levels of ICAM-1 in wounds of pressure ulcer group and diabetic foot group were significantly decreased (=16.50, 16.50, <0.01). The mRNA expression level of VCAM-1 in wounds of pressure ulcer group was significantly higher than that of diabetic foot group (=-21.50, <0.01), the mRNA expression level of ICAM-1 in wounds of pressure ulcer group was similar to that of diabetic foot group (=0, >0.05). (5) Compared with those of normal skin group, except for the mRNA expression level of IL-1β in wounds of diabetic foot group showed no significant difference (=-10.00, >0.05), the mRNA expression levels of IL-1β in wounds of burn wound group and pressure ulcer group were significantly increased (=-32.50, -21.50, <0.01); the mRNA expression levels of IL-6 were significantly increased in wounds of burn wound group, pressure ulcer group, and diabetic foot group (=-17.50, -30.50, -11.80, <0.05 or <0.01); except for the mRNA expression level of TNF-α in wounds of burn wound group showed no significant difference (=-9.50, >0.05), the mRNA expression levels of TNF-α in wounds of pressure ulcer group and diabetic foot group were significantly decreased (=18.04, 14.50, <0.01). The mRNA expression levels of IL-1β and TNF-α in wounds of pressure ulcer group were significantly lower than those of burn wound group (=11.00, 27.54, <0.05 or <0.01), while the mRNA expression level of IL-6 was significantly higher (=-13.00, <0.05). The mRNA expression levels of IL-1β and TNF-α in wounds of diabetic foot group were significantly lower than those of burn wound group (=22.50, 24.00, <0.01), while the mRNA expression level of IL-6 showed no significant difference (=5.70, >0.05). The phenotypes of diabetic foot and pressure ulcer vary from the expressions levels of proliferating cell nuclear antigen and blood vessels forming ability to the expression levels of growth factors, cell adhesion factors, and inflammatory cytokines.
比较慢性难治性伤口组织（包括压疮和糖尿病足）中炎症和修复相关因子的组织形态学及基因表达。2016年8月至2017年9月，收集南昌大学第一附属医院收治的10例泌尿外科患者（均为男性，年龄(38±4)岁）包皮环切术后的10份包皮样本作为正常皮肤组；术中收集9例热烧伤或电烧伤患者（6例男性患者，3例女性患者，年龄(51±8)岁）、13例压疮患者（9例男性患者，4例女性患者，年龄(51±14)岁）及10例糖尿病足患者（8例男性患者，2例女性患者，年龄(61±10)岁）伤口边缘有血供的组织样本。样本分为烧伤创面组（9份样本）、压疮组（13份样本）和糖尿病足组（10份样本）。分别从压疮组和糖尿病足组各取10片组织，每组5片分别采用免疫荧光法观察伤口组织形态及Ki67和CD31的表达。收集正常皮肤组10份样本、烧伤创面组9份样本、压疮组13份样本及糖尿病足组10份样本，采用实时荧光定量逆转录聚合酶链反应分析血管内皮生长因子192（VEGF192）、转化生长因子β（TGF-β）、血管细胞黏附分子-1（VCAM-1）、细胞间黏附分子-1（ICAM-1）、白细胞介素-1β（IL-1β）、IL-6及肿瘤坏死因子α（TNF-α）的mRNA表达。数据采用Mann-Whitney检验和Kruskal-Wallis秩和检验进行处理。（1）糖尿病足组Ki67表达水平（390±100）高于压疮组（182±14，=-2.611，<0.01）。（2）糖尿病足组伤口虽有大量血管内皮细胞（CD31阳性细胞），但其分布紊乱，未形成完整管腔。压疮组伤口血管内皮细胞少于糖尿病足组，但形成了完整管腔。（3）烧伤创面组、压疮组和糖尿病足组伤口VEGF192的mRNA表达水平均显著低于正常皮肤组（=13.72，30.50，15.20，<0.05或<0.01），且压疮组最低。压疮组伤口VEGF192的mRNA表达水平显著低于糖尿病足组（=15.30，<0.01）。与正常皮肤组相比，烧伤创面组伤口TGF-β的mRNA表达水平无显著差异（=-9.50，>0.05），而压疮组和糖尿病足组伤口TGF-β的mRNA表达水平显著降低（=18.04，14.50，<0.01）。压疮组伤口TGF-β的mRNA表达水平与糖尿病足组相似（=3.54，>0.05）。（4）与正常皮肤组相比，烧伤创面组和压疮组伤口VCAM-1的mRNA表达水平显著升高（=-22.50，-11.50，<0.05或<0.01），糖尿病足组伤口VCAM-1的mRNA表达水平无显著差异（=10.00，>0.05）；烧伤创面组伤口ICAM-1的mRNA表达水平无显著差异（=-9.50，>0.05），压疮组和糖尿病足组伤口ICAM-1的水平显著降低（=16.50，16.50，<0.01）。压疮组伤口VCAM-1的mRNA表达水平显著高于糖尿病足组（=-21.50，<0.01），压疮组伤口ICAM-1的mRNA表达水平与糖尿病足组相似（=0，>0.05）。（5）与正常皮肤组相比，除糖尿病足组伤口IL-1β的mRNA表达水平无显著差异（=-10.00，>0.05）外，烧伤创面组和压疮组伤口IL-1β的mRNA表达水平显著升高（=-32.50，-21.50，<0.01）；烧伤创面组、压疮组和糖尿病足组伤口IL-6的mRNA表达水平均显著升高（=-17.50，-30.50，-11.80，<0.05或<0.01）；除烧伤创面组伤口TNF-α的mRNA表达水平无显著差异（=-9.50，>0.05）外，压疮组和糖尿病足组伤口TNF-α的mRNA表达水平显著降低（=18.04，14.50，<0.01）。压疮组伤口IL-1β和TNF-α的mRNA表达水平显著低于烧伤创面组（=11.00，27.54，<0.05或<0.01），而IL-6的mRNA表达水平显著升高（=-13.00，<0.05）。糖尿病足组伤口IL-1β和TNF-α的mRNA表达水平显著低于烧伤创面组（=22.50，24.00，<0.01），而IL-6的mRNA表达水平无显著差异（=5.70，>0.05）。糖尿病足和压疮的表型在增殖细胞核抗原表达水平、血管形成能力以及生长因子、细胞黏附因子和炎性细胞因子表达水平方面存在差异。
