中文摘要：

目的探讨肝星状细胞（hepatic stellate cell，HSC）结缔组织生长因子（connective tissue growth factor，CTGF）表达对Kupffer细胞（KC）诱导的HSC激活的影响。方法构建含CTGF的RNA干扰载体。将肝星状细胞系rHSC-99分为3组：对照组、空载体组和RNA干扰组。采用RT—PCR方法检测HSC的CTGF表达。分离培养KC，建立各组HSC与KC的共培养体系，MTF检测HSC的增殖情况；RT—PCR检测HSC的TGF—β1、Ⅰ型前胶原的表达；Western Blot检测HSC的TGF—β1表达，ELISA检测共培养上清中Ⅰ型胶原的表达；免疫荧光化学检测HSC的α—SMA的表达。结果构建CTGF的RNA干扰载体Psilencer 3．1 H1-Neo—CTGF。RNA干扰后CTGF表达降低22％。KC得率为5×10^7，活率〉98％。在HSC和KC共培养体系中，RNA干扰HSC的CTGF表达后，与KC共培养的HSC活化和增殖明显被抑制，与对照组相比，HSC的增殖降低29％（t=20．17，P〈0．01）。Ⅰ型前胶原和α-SMA的表达下降38％，细胞培养上清中Ⅰ型胶原的分泌量下降48％（t=12．18，t=7．81，t=15．96，均P〈0．05）。TGF—β表达则没有明显的变化。结论阻断HSC的CTGF表达能抑制由KC诱导的HSC激活。

英文摘要：

Objective To investigate the effect of RNA interference connective tissue growth factor （CTGF） expression on Kupffer cells （KC） induced activation of hepatic stellate cells （HSC）. Methods Rat CTGF RNA interference vector Psilencer 3.1H1-Neo-CTGF was constructed and identified. HSC cell line rHSC-99 cells were divided into three groups, group A served as control, group B transfected with vector without CTGF interference, group C was RNA interference CTGF expression of HSC. RT-PCR was used to measure the expression of CTGF in HSC. Rat Kupffer cells were isolated and identified, and co-cultured with HSC in the 3 groups respectively. MTT assay was used to evaluate the proliferation of HSC. RT-PCR was used to measure the expression of TGF-β1 and precollagen type Ⅰ in HSC. Western blot was used to measure the expression of TGF-β1 in HSC. ELISA was used to detect the production of precollagen type Ⅰ protein. Immunofluorescence was used to detect the expression of α-smooth muscle actin（α-SMA） in HSC. Results After CTGF RNA interference vector transfection, CTGF expression of HSC decreased by 22% （P〈0. 01）. The yield rate of Kupffer cell was 5 × 10^7 and the cell viability exceeded 98%. In the HSC and KC co-culture system, the proliferation and activation of HSC were inhibited while RNA interference CTGF of HSC. As compared with control, HSC proliferation decreased by 29% （ P 〈 0.01 ）. Precollagen type Ⅰ and α-SMA expression decreased by 38% （P 〈 0. 01 ）. Production of precollagen type Ⅰ protein in culture medium decreased by 48% （ P 〈 0. 01 ）. Conclusions Blockade CTGF expression of HSC inhibits KC induced activation of HSC.