缺氧条件下脂多糖造成阴囊部创面不愈合的机制研究  

Mechanism of lipopolysaccharide inhibiting scrotal wound healing under hypoxic conditions

作　　者：徐娅 丛晓斌 郑怀远 刘娟 XU Ya;CONG Xiao-bin;ZHENG Huai-yuan;LIU Juan(Department of Medical Plastic Surgery,Central Hospital of Wuhan,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430014,China;Department of Medical Plastic Surgery,Wuhan Sixth Hospital,Jianghan University,Wuhan,Hubei 430030,China;Department of Hand Surgery,Wuhan Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430022,China)

机构地区：[1]华中科技大学同济医学院附属武汉中心医院医学整形美容科,湖北武汉430014 [2]江汉大学附属武汉市第六医院整形美容外科,湖北武汉430030 [3]华中科技大学附属武汉协和医院手外科,湖北武汉430022

出　　处：《中华男科学杂志》2024年第10期903-909,共7页National Journal of Andrology

基　　金：湖北省自然科学基金(2022CFC028);武汉市卫健委医学科研项目(WX20Q23)。

摘　　要：目的:研究脂多糖(LPS)对缺氧条件下阴囊皮肤来源成纤维细胞爬行能力以及其对创面愈合中血管再生能力的影响,探讨革兰阴性菌感染导致慢性阴囊区创面不愈合的潜在机制。方法:从健康供体抽取静脉血离心后的外周血血沉棕黄层分为常氧、常氧+LPS、缺氧、缺氧和LPS 4组进行预处理,分离相应培养上清,通过ELISA测定VEGF-A、FGF、IL-8分泌水平、LDH检测不同预处理上清液的细胞毒性、检测细胞迁移能力,以及人脐内皮细胞(HUVEC)的血管形成试验。结果:阴囊慢性创面内成纤维细胞在缺氧条件下表达HIF-1α的水平明显低于正常成纤维细胞(0.09±0.00 vs 0.48±0.01,P<0.05)与常氧状态相比,缺氧状态下细胞成活率无明显差异,但细胞增殖和迁移能力明显提升[(83.09±2.56)%vs(17.84±1.12)%,P<0.05];在LPS共培养条件下,阴囊成纤维细胞活性受损[存活率常氧下为%,缺氧下为(32.54±1.57)%,P<0.05],内皮细胞管形成数量明显降低(有LPS组为5.20±0.15,无LPS组为55.30±1.15,P<0.05),VEGF表达降低(有LPS组为0.59±0.07,无LPS组为1.25±0.11,P<0.05),IL-8表达增强(有LPS组为2.42±0.07,无LPS组为0.87±0.11,P<0.05)。结论:与缺氧和常氧条件相比,LPS干预的外周血血沉棕黄层来源的上清液显示出更强的细胞毒性和更少的血管生成,可能为阴囊创面难以愈合的局部重要因素。Objective:To investigate the effect of lipopolysaccharide(LPS) on the crawling ability of fibroblasts derived from the scrotal skin under hypoxic conditions,its impact on vascular regeneration during wound healing,and the potential mechanism underlying chronic non-healing scrotal wound caused by gram-negative bacterial infection.Methods:We extracted peripheral blood buffy coats from healthy donors after venous blood centrifugation,divided them into four groups(normoxia,normoxia+LPS,hypoxia,and hypoxia+LPS) for pretreatment,and isolated the corresponding cultured supernatant.Then we measured the levels of VEGF-A,FGF and IL-8 secreted using enzyme-linked immunosorbent assay(ELISA),determined the cytotoxicity of different pretreated supernatants by lactate dehydrogenase(LDH) release assay,and detected the cell migration ability and vascular formation in human-umbical vein endothelial cells(HUVEC).Results:Under hypoxic conditions,the expression of HIF-1α was significantly lower in the fibroblasts from the chronic scrotal wound than in the normal fibroblasts(0.09 ± 0.00 vs 0.48 ± 0.01,P < 0.05),the viability of the cells showed no statistically significant difference from that under normoxic conditions(P > 0.05),and the proliferation and migration abilities of the cells were markedly improved([83.09 ± 2.56]% vs [17.84 ± 1.12]%,P < 0.05).The viability of the scrotal fibroblasts was significantly lower in the normoxia+LPS than in the hypoxia+LPS group([20.15 ± 1.19]% vs [32.54 ± 1.57]%,P < 0.05).The cells treated with LPS,compared with those in the non-LPS groups,exhibited remarkably decreased endothelial cell tubes formed(5.20 ± 0.15 vs 55.30 ± 1.15,P < 0.05),down-regulated expression of VEGF(OD value:0.59 ± 0.07 vs 1.25 ± 0.11,P < 0.05) and up-regulated expression of IL-8(OD value:2.42 ± 0.07 vs 0.87 ± 0.11,P < 0.05).Conclusion:The supernatant derived from the peripheral blood buffy coats treated with LPS has a stronger cytotoxicity and weaker angiogenesis than that untreated under hypoxic or normoxic con

关 键 词：脂多糖 缺氧 阴囊皮肤来源成纤维细胞 细胞爬行 血管化 

分 类 号：R697.2[医药卫生—泌尿科学]