巨噬细胞移动抑制因子促进人增生瘢痕成纤维细胞增殖的研究

Macrophage migration inhibitory factor promotes the proliferation of human hypertrophic scar fibroblasts

目的检测巨噬细胞移动抑制因子(MIF)在人增生瘢痕组织中的表达情况,并研究MIF对人增生性瘢痕成纤维细胞(HSFBs)增殖的影响。方法实验标本来自2015年9月至2016年9月收治于长春海峡医学美容医院因增生性瘢痕(HS)自愿进行瘢痕修复手术的患者共12例,将其手术切除的标本作为HS组,其正常皮肤标本(NS)作为NS组。qRT-PCR检测MIF的基因表达,Western blot检测MIF的蛋白表达。分离原代HSFBs,使用MIF siRNA处理24 h,通过qRT-PCR检测细胞中MIF基因表达,WB检测细胞中MIF的蛋白表达以及CCK8检测细胞的增殖。结果 qRT-PCR结果显示,MIF在HS组中的表达为(0.76±0.14),NS组为(0.32±0.11),差异有统计学意义(P<0.05);Western blot检测结果显示,MIF在HS组中的表达为(0.83±0.23),NS组为(0.28±0.07),差异有统计学意义(P<0.05);成功分离HSFBs细胞后,通过siRNA技术抑制MIF的表达,CCK8结果显示HSFBs+MIF siRNA组吸光度为(0.25±0.023),HSFBs组吸光度为(0.45±0.03),HSFBs+siRNA NC组吸光度为(0.42±0.04),HSFBs+MIF siRNA组HSFBs增殖能力较HSFBs组及HSFBs+siRNA NC组明显降低,差异均有统计学意义(P<0.05)。结论 MIF在增生性瘢痕中的表达明显高于正常皮肤,且MIF对HS成纤维细胞的增殖有促进作用。

Objective To detect the expression of macrophage migration inhibitory factor（MIF） in human hypertrophic scar tissue, and to study the effect of MIF on proliferation of human hypertrophic scar fibroblasts（HSFBs）.Methods The experimental specimens were collected from twelve patients in Changchun Strait Medical Beauty Hospital between September 2015 to September 2016. The twelve patients volunteered for scar repair surgery because of hypertrophic scar（HS）. The specimens removed surgically were used as HS group, and the specimens from normal skins were used as NS group. qRT-PCR assay was used to detect the gene expression level of MIF, and Western blot assay was used to measure the protein expression level of MIF. Primary HSFBs were isolated, and transfected with MIF siRNA for24 h. The gene expression level of MIF was detected by qRT-PCR assay, and the protein expression level of MIF was measured by Western blot assay. The cell proliferation ability was detected by CCK8 assay. Results qRT-PCR assay showed that the expression level of MIF in the HS group was（0.76±0.14）, significnatly higher than（0.32±0.11） in the NS group（P〈0.05）. Western blot assay showed that the protein expression level of MIF was（0.83 ± 0.23）, significant higher than（0.28±0.07） in the NS group（P〈0.05）. After successful isolation of HSFBs cells, siRNA was used to inhibit the expression level of MIF in HSFBs. CCK8 assay showed that the absorbance in the HSFBs＋MIF siRNA group was（0.25±0.023）, versus（0.45±0.03） in HSFBs group and（0.42±0.04） in HSFBs＋ siRNA NC group, indicating that cell proliferation was significantly decreased in the MIF siRNA transfected cells compared with blank and negative control cells（P〈0.05）. Conclusion The expression level of MIF in hypertrophic scar is significantly higher than that in normal skin,and MIF promotes the proliferation ability of HSFBs.