中文摘要：

目的观察半乳糖凝集素-3（galectin-3）表达与肝星状细胞增殖和凋亡的关系。方法采用逆转录聚合酶链反应（RTPCR）和Western blot检测大鼠肝星状细胞中galectin-3 mRNA和蛋白的表达；设计合成galectin-3小干扰片段，筛选有效片段构建质粒；转染大鼠肝星状细胞系rHSC-99，并设立对照组和空质粒转染组，RTPCR及Western blot检测RNA干扰后galectin-3的表达。活细胞计数法观察干扰后24、48、72h各组细胞增殖情况；Annexin-V／PI染色，流式细胞仪检测转染72h后各组细胞凋亡情况。组间比较用LSD法t检验进行统计学分析。结果rHSC-99细胞表达galectin-3；成功构建pGCsilencer U6／Neo／GFP／shRNA，并经测序验证正确。转染效率为40％～50％，RT-PCR结果显示：RNA干扰组galectin-3 mRNA表达量较对照组下降了52．4％（F=136．432，P〈0．01），Western blot显示RNA干扰组galectin-3蛋白表达量较对照组下降了69．2％（F=144．842，P〈0．01）。干扰后48h和72h时，galectin-3 RNA干扰组细胞增殖较对照组分别下降38．1％（F=4．425，P=0．010）和37．5％（F=4．415，P=0．018）；干扰72h时，galectin-3 RNA干扰组细胞凋亡率显著高于对照组：早期凋亡率增加155．2％，与对照组比较，F=27．588，P〈0．01；晚期凋亡率增加56．9％，与对照组比较，F=12．798，P〈0．01。结论rHSC-99细胞表达galecitn-3；RNA干扰rHSC-99细胞galectin-3的表达能抑制rHSC99的增殖并促进其凋亡。

英文摘要：

Objective To observe the effects of galectin-3 on proliferation and apoptosis of hepatic stellate cells. Methods RT-PCR and Western blot were used to detect the expression of galectin-3 in hepatic stellate cells. Short hairpin DNA targeting galectin-3 of rat was was ligated into the recombinant vector pGCsilencer U6/Neo/GFP/shRNA plasmid. Then the plasmid was transfected into rat hepatic stellate cells. RT-PCR and Western blot were used to detect the interfering efficiency. Cell proliferation level was observed by CCK8 method at 24, 48 and 72 hours after transfection. Cell apoptosis was measured by Annexin V/PI- labeled flow cytometric analysis. Results Expression of galectin-3 in HSC was verified by both RT-PCR and Western blot. The recombinant vector was successfully constructed and verified, and was transfected into rat hepatic stellate cells. Western Blot and RT-PCR results demonstrated that the expression level of Galectin-3 was significantly down-regulated in galectin-3 shRNA transfected cells compared to control vector transfeced cells. CCK8 assay indicated that proliferation of Galectin-3 knockdown cells was lower than that of control cells 48 and 72 hours post-transfection. Apoptotic cells in shRNA-interfering group were higher than those in control group both in early stage and advanced stage. Conclusion Hepatic stellate cells can express galectin-3. Inhibition of galectin-3 using RNAi technique can suppress proliferation and induce apoptosis in HSC.