Mechanism of lipopolysaccharide inhibiting scrotal wound healing under hypoxic conditions
XU Ya
CONG Xiao-bin
ZHENG Huai-yuan
LIU Juan
Abstract：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 un-derlying 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 superna-tants 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-1 α was significantly lower in the fibro-blasts 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 endo-thelial cell tubes formed(5.20±0.15 va 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 an-giogenesis than that untreated under hypoxic or normoxic conditions,which may be one of the critical factors for difficult healing of scrotal wound.
Keywords：lipopolysaccharidehypoxiafibroblasts derived from scrotal skincell crawlingangiogenesis
Publication Date：2024-10-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 903-909 )