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本文旨在研究川芎嗪及大鼠结缔组织生长因子(CTGF)miRNA质粒对高糖刺激大鼠肝星状细胞(HSC)的CTGF和转化生长因子-beta(TGF-beta)mRNA表达量、蛋白表达量及Ⅰ型胶原表达量的作用。首先体外培养大鼠HSC以及稳定表达大鼠CTGF miRNA和稳定表达阴性质粒的大鼠HSC,高糖及川芎嗪作用后提取总RNA、总蛋白及收集细胞上清液,检测川芎嗪及大鼠CTGF miRNA质粒对高糖刺激HSC的CTGF、TGF-beta及Ⅰ型胶原表达量。实验结果表明,高糖导致HSC的CTGF mRNA、CTGF蛋白、TGF-beta mRNA、TGF-beta蛋白及Ⅰ型胶原表达量增加(P<0.05)。川芎嗪组CTGF mRNA、CTGF蛋白、TGF-beta mRNA、TGF-beta蛋白及Ⅰ型胶原的表达量较高糖组低(P<0.05)。大鼠CTGF miRNA质粒干扰组CTGF mRNA、CTGF蛋白、TGF-beta mRNA、TGF-beta蛋白及Ⅰ型胶原的表达量较高糖组低(P<0.05)。川芎嗪组与大鼠CTGF miRNA质粒干扰组相比,CTGF mRNA及CTGF蛋白的表达量差异无统计学意义(P>0.05),川芎嗪组Ⅰ型胶原的表达量明显低于大鼠CTGF miRNA质粒干扰组(P<0.05)。因此,高糖可刺激CTGF、TGF-beta、Ⅰ型胶原的表达,而川芎嗪、大鼠CTGF miRNA质粒可导致CTGF、TGF-beta、Ⅰ型胶原的表达量下降。
The aim of this study was to investigate the effects of ligustrazine and rat CTGF miRNA plasmids on the expression of CTGF and TGF-beta mRNA in high-glucose-stimulated rat hepatic stellate cells (HSC) Amount and type Ⅰ collagen expression role. First of all, rat HSC was cultured in vitro and rat HSC stably expressing rat CTGF miRNA and negative plasmid were stably transfected. After treated with high glucose and ligustrazine, total RNA, total protein and supernatant were collected to detect the effects of ligustrazine and rat CTGF Expression of CTGF, TGF-beta and type I collagen by miRNA plasmid in HSC stimulated by high glucose. The experimental results showed that the expression of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta protein and type I collagen in HSC increased (P <0.05). The expression of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta protein and type I collagen in ligustrazine group was lower than that in high glucose group (P <0.05). The expression of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta protein and type I collagen in CTGF miRNA plasmid interference group was lower than that in high glucose group (P <0.05). Compared with CTGF miRNA plasmid interference group, the expression of CTGF mRNA and CTGF protein in ligustrazine group was not significantly different (P> 0.05). The expression of collagen type Ⅰ in ligustrazine group was significantly lower than that in CTGF miRNA plasmid Interference group (P <0.05). Therefore, high glucose could stimulate the expression of CTGF, TGF-beta and type I collagen, while tetramethylpyrazine and rat CTGF miRNA plasmid could decrease the expression of CTGF, TGF-beta and type I collagen.