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细胞外基质(ECM)积聚与活化肝星状细胞(HSC)数量密切相关,增殖是活化HSC的重要生物学特性之一,也是调控活化 HSC数量的一种重要方式。结缔组织生长因子(CTGF)主要由活化HSC产生,是HSC增殖的一个重要刺激因子。因此, 下调CTGF表达,将有望通过抑制HSC增殖,减少活化HSC 数量及ECM积聚防治肝纤维化发生与发展。小分子干扰RNA (siRNA)是一种转录后基因沉默的强大工具。研究旨在探讨是否抗CTGF siRNA能下调原代HSC CTGF基因表达及抑制 HSC增殖。一、材料与方法 1．材料：SD大鼠购自中国科学院上海实验动物中心,大鼠CTGF siRNA(序列483～503)由中国科学院上海生命研究院生物化学与细胞研究所合成,Oligofectamine及Opti-MEM1 为美国Invitrogen公司产品；Ⅳ型胶原酶、Nycodenz及小鼠
The accumulation of extracellular matrix (ECM) is closely related to the number of activated hepatic stellate cells (HSCs). Proliferation is one of the important biological characteristics of activated HSCs and an important way to regulate the number of activated HSCs. Connective tissue growth factor (CTGF) is mainly produced by activated HSC and is an important stimulator of HSC proliferation. Therefore, downregulation of CTGF expression is expected to prevent and treat the occurrence and development of liver fibrosis by inhibiting HSC proliferation, reducing the number of activated HSCs and accumulating ECM. Small interfering RNA (siRNA) is a powerful tool for post-transcriptional gene silencing. The aim of this study was to investigate whether anti-CTGF siRNA down-regulates primary HSC CTGF gene expression and inhibits HSC proliferation. First, materials and methods 1. MATERIALS: SD rats were purchased from Shanghai Experimental Animal Center, Chinese Academy of Sciences. Rat CTGF siRNA (sequences 483-503) was synthesized by the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. Oligofectamine and Opti-MEM1 were purchased from Invitrogen Corporation. Type IV collagenase, Nycodenz and mice