CCL2促进肝再生中细胞外基质的形成

CCL2 Promotes the Formation of ExtracellularMatrix in Liver Regeneration

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.

To study the function of chemokine（ C-C motif） ligand 2 （ CCL2） during liver regeneration（LR）,we constructed an expression vector （CCL2-N1） and an interference vector[CCL2 （255）]. The expression peak of endogenous CCL2 was at 72 hours after partial hepatectomy （PH） determined by Rat Genome 230 microarray V2. 0,real-time PCR and Western blot. The expression of exogenous CCL2 fused green fluorescent protein （ GFP） was peaked at 24 hours after transfection. Therefore,the CCL2-N1 plasmids were introduced into rats at 48 hours after PH was chosen to coordinate with the function of endogenous CCL2. The value of liver coefficient and the expression of proliferating cell nuclear antigen（PCNA） in transfected rats were significantly higher than those of the controls. The levels of hyaluronicacid and laminin were also significantly increased. Conversely,interference plasmid CCL2 （ 255 ） reduced the liver coefficient and PCNA expression,and significantly decreased procollagen Ⅲ peptide,Ⅳ collagen,hyaluronic acid and laminin. In conclusion,CCL2 might promote liver regeneration throughregulating the synthesis of extracellular matrix.