缺氧对瘢痕成纤维细胞缺氧诱导因子1α表达的影响

Effects of hypoxia on the expression of hypoxia inducible factor 1 alpha in scar fibroblasts

目的:探讨缺氧程度与瘢痕成纤维细胞缺氧诱导因子1α表达的关系。方法:实验于2004-01/03在湘雅医学院中心实验室完成。选择烧伤患者创面愈合后3～6个月的瘢痕为取材对象。取体外分离培养的皮肤瘢痕成纤维细胞,分别在体积分数为0.2,0.1,0.05,0.015的氧浓度下培养24h后,采用免疫组织化学染色和WesternBlot方法检测缺氧诱导因子1α的表达。结果:①免疫组织化学染色:细胞核和细胞质均有缺氧诱导因子1α表达,体积分数为0.2,0.1,0.05,0.015的氧浓度下缺氧诱导因子1α表达的信号强度分别为47.50±9.29,96.00±18.29,209.17±19.64,263.83±13.24,各浓度比较差异显著(F=261.01,P<0.01)。体积分数为0.2的氧浓度下缺氧诱导因子1α部分为弱阳性,体积分数为0.1的氧浓度下缺氧诱导因子1α基本弱阳性,体积分数为0.05的氧浓度下缺氧诱导因子1α以阳性表达为主,体积分数为0.015的氧浓度下缺氧诱导因子1α基本为阳性。②WesternBlot印迹检测:体积分数为0.2,0.1,0.05,0.015的氧浓度下缺氧诱导因子1α蛋白表达相对含量分别为0.02,0.26,0.39,0.97,呈逐渐增高趋势,均明显高于体积分数为0.2的氧浓度条件下的缺氧诱导因子1α蛋白表达。结论:随着氧浓度的降低,皮肤瘢痕成纤维细胞的缺氧诱导因子1α表达强度逐渐增强。缺氧诱导因子1α的表达强度可间接反映瘢痕组织内的氧浓度。

AIM： To explore relationship between extent of hypoxia and exprssion of hypoxia inducible factor 1α（HIF-1α） in scar fibroblasts. METHODS： The experiment was conducted in the Central Laboratory of Xiangya Hospital from January to March 2004. Cicatricles of burnt patients 3-6 months after healed were taken as subjects. Scar fibroblasts isolately cultured in vitro were inspected with immunohistochemical staining and Western Blot for expression of HIF-1α 24 hours after cultured with the volume fraction of theoxygen concentration of 0.2, 0.1, 0.05 and 0.015. RESUTLS： ①Immunohistochemical staining： There were expressions of HIF-1α in cellular nucleus and cell-substance, signal intensities of expression of HIF-1α under the volume fraction of theoxygen concentration of 0.2, 0.1, 0.05 and 0.015 were respectively 47.50±9.29, 96.00±18.29, 209.17 ±19.64 and 263.83 ±13.24,differences of comparion in different concentrations were significant （F=261.01, P 〈 0.01）. Partial expressions of HIF-1α were weakly positive （WP） in 0.2 O2 and were fundamentally in WP in 0.1 O2. While expressions of HIF-1α in 0.5 O2 were mainly positive and that in 0.015 O2 were fundamentally positive. ② Western Blot inspection： Relative amounts of protein expression of HIF-1α in 0.2, 0.1, 0.05, 0.015 O2 were respectively 0.02,0.26,0.39 and 0.97 ,which were in increasing tendency and higher than that in 0.2 O2. CONCLUSION： With the decreasing of O2 concentration, expression of HIF-1α in scar fibroblasts is strengthened gradually. Expressive intensity of HIF-1α can indirectly reflect the concentration of O2 in scar tissues.