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目的检测中药髓复康(SFK)对脊髓损伤早期相关基因谱表达的调节作用。方法86只成年雄性SD大鼠,随机分为3组,即SFK治疗组(S组)35只,脊髓损伤模型组(M组)和正常对照组(N组)。按双盲法原则制备大鼠T12脊髓右侧半横断损伤(SCI)模型。造模后24h取出损伤区的脊髓组织。经提取总RNA、纯化的mRNA、逆转录合成荧光标记的cDNA杂交探针、与Biostar40s组织基因芯片杂交、对杂交信号的分析和组间比较等步骤,确定差异表达(上调或下调)基因。结果M组与N组比较,有139条差异表达基因,其中83条上调,56条下调。这些差异表达的基因功能主要涉及细胞代谢,特别是脂类代谢的调节、离子通道的调节、免疫和防御功能的调节、细胞骨架蛋白和细胞外基质、细胞周期和生长的调节、细胞信号传导和基因转录的调节等。M组与S组比较,有37条差异表达基因,有11条上调,26条下调。其中Anxa1、Ca2+-ATPase、IL-1β、ApoCⅢ和PFP基因上调;HSP-70和Psap下调。这些基因与神经组织保护、生长和再生关系密切。结论SCI的早期有139条基因的表达发生了显著的变化,差异表达基因的功能是多方面的,与SCI后神经组织的变性坏死、创伤性炎症和继发性脊髓损伤的进展和修复再生能力的丧失等有关。SFK可以调节SCI诱导的基因表达变化,这种调整作用不仅可以启动神经组织保护作用,而且也在一定的程度上激发和支持着损伤神经组织(包括神经纤维)的再生。
Objective To investigate the regulatory effect of traditional Chinese medicine fulukoneukang (SFK) on the expression of related genes in the early stage of spinal cord injury. Methods Eighty-six adult male Sprague-Dawley rats were randomly divided into 3 groups: SFK treatment group (S group), SCI group (M group) and normal control group (N group). According to the double-blind principle, the rat model of spinal cord right hemisection (SCI) was prepared. 24 hours after the modeling, the spinal cord tissue in the injured area was removed. The differentially expressed (up- or down-regulated) genes were identified by extraction of total RNA, purified mRNA, reverse transcription synthesis of fluorescently labeled cDNA hybridization probes, hybridization with Biostar 40s tissue microarrays, analysis of hybridization signals, and comparisons between groups. Results There were 139 differentially expressed genes in M group compared with N group, of which 83 were up-regulated and 56 were down-regulated. These differentially expressed gene functions are mainly involved in cell metabolism, especially regulation of lipid metabolism, regulation of ion channels, regulation of immune and defense functions, regulation of cytoskeletal proteins and extracellular matrix, cell cycle and growth, cell signaling, and Gene transcription regulation and so on. There were 37 differentially expressed genes in M group compared with S group, with 11 up-regulated and 26 down-regulated. Among them, Anxa1, Ca2+-ATPase, IL-1β, ApoCIII and PFP genes were up-regulated; HSP-70 and Psap were down-regulated. These genes are closely related to the protection, growth and regeneration of nervous tissues. Conclusion The expression of 139 genes in the early stage of SCI has undergone significant changes. The functions of differentially expressed genes are multifaceted. Degeneration and necrosis of nervous tissue, progression of traumatic inflammation and secondary spinal cord injury and repair and regeneration after SCI Loss and other related. SFK can regulate SCI-induced changes in gene expression. This regulation not only initiates neuroprotection, but also to a certain extent stimulates and supports the regeneration of damaged neural tissue (including nerve fibers).