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为阐明CC趋化因子配体2〔chemokine(C-C motif)ligand 2,CCL2〕对肝再生(liverregeneration,LR)的影响,构建CCL2的表达载体CCL2-N1及其干涉载体CCL2(255).为确定合适的转基因时间,观测内外源性CCL2的表达高峰.Rat Genome 230 2.0芯片、实时定量PCR和Western印迹显示,内源性CCL2的表达高峰在部分肝切除(partial hepatectomy,PH)后72 h.与CCL2融合表达的绿色荧光蛋白(green fluorescent protein,GFP)的表达和实时定量PCR显示,外源性CCL2的表达高峰在转基因后的24 h.所以,为使内源性和外源性CCL2的作用叠加,选择PH后48 h进行转基因.此时将CCL2-N1质粒转入大鼠体内,发现转基因后大鼠肝系数、增殖细胞核抗原(PCNA)表达量显著高于对照,透明质酸和层粘连蛋白含量显著升高.相反,干涉质粒CCL2(255)能降低肝系数,减少PCNA表达量,并且Ⅲ型前胶原肽、Ⅳ型胶原、透明质酸和层粘连蛋白含量与对照相比,有显著或极显著降低.综上所述,CCL2是肝再生相关基因,它能提高肝系数,可能通过调节细胞外基质(extracellular matrix,ECM)合成而促进肝脏再生.
CCL2 expression vector CCL2-N1 and its interference vector CCL2 (255) were constructed in order to elucidate the effect of chemokine CC receptor2 (CCL2) on liver regeneration (LR) Appropriate gene transfer time was used to observe the peak of endogenous and exogenous CCL2 expression.Rat Genome 230 2.0 chip, real-time PCR and Western blotting showed that the expression of endogenous CCL2 peaked at 72 h after partial hepatectomy (PH) CCL2 fusion expression of green fluorescent protein (green fluorescent protein, GFP) expression and real-time quantitative PCR showed that exogenous CCL2 expression peak at 24 h after transgene, therefore, in order to endogenous and exogenous CCL2 role Superposed, and selected for 48 h after transfection, the CCL2-N1 plasmids were transferred into rats and found that the transgenic rat liver coefficient and proliferating cell nuclear antigen (PCNA) expression was significantly higher than the control, hyaluronic acid and lamina In contrast, CCL2 (255) decreased the hepatic coefficient and decreased the expression of PCNA, and the content of type III procollagen peptide, type IV collagen, hyaluronic acid and laminin was significantly higher than that of the control Or significantly reduced. The, CCL2 gene related liver regeneration, it can improve liver coefficients, may promote liver regeneration by regulating the extracellular matrix (extracellular matrix, ECM) synthesis.