【摘要】 目的:探讨重组腺相关病毒介导的IL-32基因抑制人瘢痕疙瘩成纤维细胞(Keloid fibroblasts,KFs)增殖的机制。方法:用重组腺相关病毒介导的IL-32(rAAV2IL-32)及腺相关病毒空载体感染人KFs,通过荧光显微镜观察rAAV2IL-32的感染效率,RT-PCR法检测IL-32的转录水平;用MTT法检测KFs增殖的情况;用蛋白质印迹方法检测Bcl-2、BAX、TGF-β1的表达。结果:rAAV2IL-32及腺相关病毒空载体感染KFs 48h后,RT-PCR检测显示rAAV2IL-32组出现高表达电泳条带,而腺相关病毒空载体组则未出现电泳条带;MTT法检测显示r AAV2I L-32组明显抑制了KFs的增殖;蛋白质印迹结果显示rAAV2IL-32组KFs中Bcl-2、TGF-β1表达降低,BAX表达增高。结论:rAAV2IL-32可能是通过减少KFs中Bcl-2和TGF-β1的表达以及增加BAX的表达来抑制瘢痕疙瘩的增殖。更多还原

【Abstract】 Objective Explore the mechanism of IL-32 gene regulation in Keloid fibroblasts(KFs).Methods With adeno-associated virus-mediated IL-32(rAAV2 IL-32) and empty vector of adeno-associated virus infection of human KFs.Infection efficiency of adeno-associated virus was observed by fluorescence microscopy.Transcription levels of IL-32 was detected by RT-PCR assay.The effects of IL-32 on KFs cell proliferation was detected by MTT.Bcl,BAX,TGF-β1 expression was detected by Western blot.Results After of rAAV2 IL-32 and adeno-associated virus empty vector infection KFs 48h,RT-PCR test results showed rAAV2 IL-32 group of high expression of electrophoretic bands,adeno-associated virus vector group did not appear electrophoretic bands.MTT assay showed that of rAAV2 IL-32 significantly inhibited the KFs proliferation.Western blot results showed that overexpression of Bcl IL-32 KFs,reduced expression of TGF-β1,increased expression of BAX.Conclusion IL-32 to inhibit proliferation of scar may reduce the expression of Bcl and TGF-β1 and increased expression of BAX.更多还原