川芎嗪及大鼠CTGF miRNA质粒对高糖刺激肝星状细胞结缔组织生长因子、转化生长因子-beta的影响

Effect of Tetramethylpyrazine and Rat CTGF miRNA Plasmids on Connective Tissue Growth Factor,Transforming Growth Factor-beta in High Glucose Stimulated Hepatic Stellate Cells

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。

Abstract： The aim of this research is to evaluate the effect of tetramethylpyrazine （TMP） and connective tissue growth factor （CTGF） miRNA plasmids on the expressive levels of CTGF, transforming growth factor-beta （TGF- beta） and type I collagen of rat hepatic stellate cells （HSC） which are stimulated by high glucose. The rat HSCs which were successfully transfected rat CTGF miRNA plasmids and the rat HSCs which were successfully transfect- ed negative plasmids were cultured in vitro. After stimulus of the TMP and the high glucose, the protein levels and gene expressive levels of CTGF, TGF-beta and type I collagen were tested. The results indicated that high glucose increased the expression of CTGF mRNA, CTGF protein, TGF-heta mRNA,TGF-beta protein and type I collagen （P〈0.05）. The expressive levels of CTGF mRNA, CTGF protein, TGF-beta mRNA, TGF-beta and type I colla- gen in TMP group were lower than those in high glucose group and showed statistically significant differences （P〈0.05）. Compared with high glucose group, the expressive levels of CTGF mRNA, CTGF protein, TGF-beta mR- NA, TGF-beta and type I collagen in rat CTGF miRNA plasmid interference group were significantly lower （P〈 0.05）. However, no statistically significant differenee was found in CTGF mRNA and CTGF protein levels between TMP group and CTGF miRNA group （P〉0. 05）, while type I collagen levels showed statistically significant differences （P〈0.05）. It is concluded that high glucose could promote the expressions of CTGF, TGF-beta and type I collagen, and TMP and rat CTGF miRNA plasmids could reduce the expressions of CTGF, TGF-beta, type I collagen.