HIF-1α对病理性瘢痕疙瘩成纤维细胞增殖活力的影响及机制

摘要

Objective To explore the effect of hypoxia-inducible factor 1α(HIF-1α)in pathological scar and its specific mechanism,and the therapy target of pathological scar.Methods Real time PCR and Western blot was used to test the expression of HIF-1αin normal tissue and pathological scar,meanwhile to detect the effect of hypoxic environment on the expression of HIF-1α.Fibroblasts activity in pathological scar and normal tissue under different oxygen concentration(20%,10%,5%,2% and 1%)was determined by MTT method.Determine the different expression of HIF-1αmRNA and protein in normal environment(20% oxygen) and hypoxic environment(5%oxygen).The changes of the number of fibroblasts after the silence of HIF-1αby HIF-1αshRNA virus using flow cytometer.Results The Real time PCR and Western blot results showed that the expressions of HIF-1αmRNA and protein in fibroblasts of pathological scar was significantly higher than that in normal tissue(P<0.05),and the activity of the keloid fibroblasts in hypoxia environment(5%,2% and 1%)was also higher than that in normal tissue(P<0.05).The expressions of HIF-1αmRNA and protein in keloid fiber cells were higher in hypoxia environment(5% oxygen)than those in normal environment(20% oxygen),but when silence the expression of HIF-1αin keloid fibroblasts,the apoptosis of fibroblast in hypoxia environment(5%oxygen)were significantly higher than control group[(0.021 ±0.001)%vs.(3.739 ± 0.039)%,P<0.05].Conclusion HIF-1αincreases significantly in the pathological keloid fibroblasts and could promote the proliferation of fibroblasts and its vitality,which could accelerate the formation of pathological scar.%目的：探究低氧诱导因子（ hypoxia-inducible factor 1α，HIF-1α）在病理性瘢痕疙瘩发生中的具体作用机制，寻找瘢痕疙瘩治疗的重要靶位点。方法 Real time PCR和Western blot检测正常组织和瘢痕疙瘩中HIF-1α的mRNA及蛋白表达。 MTT法检测不同氧浓度环境下（20％、10％、5％、2％和1％）正常组织和瘢痕疙瘩中成纤维细胞的活性，检测瘢痕疙瘩在5％低氧环境与20％正常氧环境下HIF-1α的mRNA及蛋白表达。 HIF-1αshRNA病毒沉默HIF-1α后采用流式细胞仪检测5％低氧环境下瘢痕疙瘩中成纤维细胞的数量变化。结果 Real time PCR和Western blot检测结果显示瘢痕疙瘩成纤维细胞中HIF-1α的表达较正常组织明显升高（P＜0.05），且在低氧环境中（氧浓度5％、2％和1％）的成纤维细胞活性较正常组织显著升高（P＜0.05）。瘢痕疙瘩中成纤维细胞中HIF-1α的mRNA及蛋白表达在5％低氧环境显著高于20％正常氧环境（P＜0.05）。当沉默瘢痕疙瘩中HIF-1α的表达后，5％低氧环境中的成纤维细胞凋亡数量显著高于对照组[（0.021±0.001）％vs.（3.739±0.039）％，P＜0.05]。结论 HIF-1α在病理性瘢痕疙瘩成纤维细胞中显著升高且促进成纤维细胞的增殖及活力，对瘢痕疙瘩的形成具有一定的促进作用。