人工合成IKVAV多肽对星形胶质细胞增殖和Neurocan基因表达的影响  

Effects of man-made IKVAV polypeptide on astrocyte proliferation and gene expression of Neurocan

作　　者：王天瑞[1] 马学晓[1] 于腾波[1] 相宏飞[1] 陈伯华[1] 

机构地区：[1]青岛大学医学院附属医院脊柱外科,山东266003

出　　处：《脊柱外科杂志》2012年第2期109-112,共4页Journal of Spinal Surgery

基　　金：山东省青年科学家研究奖励基金(2007BS03047);青岛市科技发展计划(06-2-2-6 nsh-2)

摘　　要：目的观察人工合成IKVAV多肽对星形胶质细胞增殖及硫酸软骨素蛋白多糖核心蛋白Neurocan基因表达的影响,并探讨IKVAV潜在的促进受损脊髓功能恢复的作用。方法设计IKVAV序列,人工合成IKVAV多肽,将星形胶质细胞接种于1%IKVAV包被的培养板上,分别于培养1 d、3 d、5 d、7 d、9 d后在激光共聚焦显微镜下观察IKVAV多肽和星形胶质细胞的组织相容性,用CCK-8法检测星形胶质细胞增殖情况,用荧光定量PCR法检测星形胶质细胞硫酸软骨素蛋白多糖核心蛋白Neurocan的表达,并与对照组比较,进行统计学分析。结果与对照组相比,实验组培养板上星形胶质细胞总数明显减少,但细胞存活率>95%。荧光定量PCR结果显示实验组星形胶质细胞Neu-rocan的表达量较对照组显著降低。结论人工合成IKVAV多肽可抑制星形胶质细胞增殖、下调硫酸硫酸软骨素蛋白多糖的表达,从而抑制受损脊髓胶质瘢痕的形成,促进脊髓功能恢复。Objective To observe the effects of man-made isoleucine-lysine-valine-alanine-valine（IKVAV） polypeptide on the astrocyte proliferation and the gene expression of Neurocan [core protein of chondroitin sulfate proteoglycans（CSPGs）],and to explore the potential role of IKVAV in the functional recovery of the damaged spinal cord.Methods IKVAV sequence was designed and the peptide was synthesized.Astrocytes were inoculated at 1% IKVAV-coated culture plates.The histocompatibility of IKVAV polypeptide and astrocytes was observed with laser confocal microscopy 1 d,3 d,5 d,7 d,9 d after inoculation.The proliferation of astrocytes and Neurocan expression were detected by CCK-8 and fluorescent quantitative PCR（FQ-PCR）,respectively,at the same time points.Then the results were statistically analyzed and compared with control group.Results Compared with control group,astrocytes at IKVAV-coated culture plate were significantly reduced,but the survival rate was more than 95%.FQ-PCR results showed that Neurocan expression in IKVAV-coated astrocytes was significantly lower than that in control group.ConclusionMan-made IKVAV polypeptide can inhibit astrocyte proliferation and downregulate CSPGs expression.Consequently,it can inhibit glial scar formation and promote functional recovery of spinal cord.

关 键 词：肽类 星形细胞 基因表达 细胞增殖 脊髓损伤 

分 类 号：R651.21[医药卫生—外科学]