反义结缔组织生长因子对人瘢痕疙瘩成纤维细胞的作用
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[摘要]目的： 研究结缔组织生长因子(CTGF)反义寡核苷酸(ASODN)对人瘢痕疙瘩成纤维细胞CTGFmRNA的表达和细胞增殖及胶原合成的影响，探讨瘢痕疙瘩的基因治疗。方法：体外分离、培养人正常皮肤成纤维细胞(NSF)和瘢痕疙瘩成纤维细胞(KF)，以脂质体介导方法将CTGFASODN转染KF中，用逆转录-聚合酶链式反应(RT-PCR)方法检测细胞中CTGFmRNA的表达；采用MTT方法测细胞增殖；3H-脯氨酸掺入法检测细胞的胶原合成量。 结果：CTGFmRNA在NSF中几乎无表达，在KF中表达增高，CTGFASODN可以抑制其增殖和胶原合成(P<0.05)。结论：CTGFASODN能够抑制成纤维细胞CTGFmRNA表达和细胞增殖及胶原合成，表明阻断CTGF可能是延缓瘢痕纤维化的有效手段。
[关键词]结缔组织生长因子；反义寡核苷酸；瘢痕疙瘩
[中图分类号]R619+.6[文献标识码]A[文章编号]1008-6455(2011)06-0936-03
Effects of connective tissue growth factor antisense oligonucleotides on the cultured human keloid fibroblasts
REN Li-hong1, DUAN Guo-xin2, YANG Da-ping1, LIU Ying1, XIAO Zhi-bo1, TENG Wen1, LIU Guo-feng1
(1.Plastic Cosmetic Center,The Second Affiliated Hospital of Harbin Medical University,Harbin 150086,Heilongjiang,China; 2.Department of Plastic Surgery,Shengmeijia Plastic Surgery Clinic,Shenzhen 518000,Guangdong,China)
Abstract:ObjectiveTo explore the effects of connective tissue growth factor(CTGF) antisense oligonucleotides on the expression of CTGF mRNA, cell proliferation and collagen synthesis in the cultured human keloid fibroblasts. MethodsNormal skin fibroblasts(NSF) and keloid fibroblasts(KF) were cultured in vitro, CTGF antisense oligonucleotides were transfected into the KFs by liposome. The expression of CTGFmRNA was assessed by reverse transcription polymerase chain reaction method(RT-PCR).Cell proliferation was detected by MTT method and the collagen synthesis of KF was assessed by 3H-proline incorporation method.ResultsCTGFmRNA was strongly expressed in KF(P<0.05), whereas it was not expressed in NSF. CTGF antisense oligonucleotides could inhibited CTGFmRNA expression, cell proliferation and collagen synthesis.ConclusionsCTGF antisense oligonucleotides could inhibited the expression of CTGFmRNA, cell proliferation and collagen synthesis, implying that CTGF may be a potential therapeutic target to scar fibrosis.
Key words: connective tissue growth factor; antisense oligonucleotides; keloid
瘢痕疙瘩(Keloid)是以成纤维细胞为主的细胞成分过度增殖和以胶原为主的细胞外基质过度沉积的皮肤纤维化疾病[1] ......
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