Objective To evaluate the effect of silencing leptin by small interfering RNA(siRNA)on the expression of lep‐tin ,and apoptosis ,proliferation and intracellular Ca2+ concentration([Ca2+ ]i )of hepatic stellate cells(HSCs)and to provide evi‐dence for liver fibrosis gene therapy.Methods HSCs were divided into normal control group ,blank vector group ,siRNA nega‐tive control group and leptin‐siRNA group.After transfection of the leptin‐siRNAs into HSCs ,cell proliferation was measured by MTT assay.Cell cycle and apoptosis were measured by flow cytometry.Expression of leptin was detected by immunocyto‐chemistry and Western blot. [Ca2+ ]i was measured by Fura‐2/AM loading.Results Compared with the normal control group , the blank vector group and the siRNA control group ,the protein expression of leptin and the cell growth were significantly in‐hibited in the leptin‐siRNA group(P<0.05). The proliferation rate of HSCs was significantly different at different time points (24 ,48 and 72 h)(P<0.05).The cell apoptosis rate was increased significantly in the leptin‐siRNA group(P<0.01).At the same time ,Leptin‐siRNA‐induced [Ca2+ ]i was also significantly reduced(P<0.05).Conclusion The leptin gene may play an important role in liver fibrosis progression and is potentially a novel predictive and prognostic marker for liver fibrosis.%目的：探讨靶向瘦素（leptin）基因的小干扰RNA（small interfering RNA ，siRNA）对leptin表达，肝星状细胞（hepatic stellate cell ，HSC）的增殖、凋亡及细胞内钙离子浓度（intracellular Ca2＋ concentration ，[Ca2＋]i ）的影响，为肝纤维化基因治疗提供可行的方案。方法 HSC分为4组：正常对照组、空载体组、siRNA对照组、leptin‐siRNA组。依分组要求分别转染 HSC后，采用M T T法和流式细胞技术检测各组细胞的凋亡和增殖；应用免疫细胞化学SP法、Western blot法检测leptin蛋白的表达；Fura‐2／AM负载检测[Ca2＋]i变化。结果与HSC的正常对照组、空载体组及siRNA对照组相比，leptin‐siRNA组可以有效抑制leptin基因的表达和细胞增殖（P＜0.05），且不同时段（24、48、72 h）细胞抑制率之间有显著差异（P＜0.05）。leptin‐siRNA组细胞凋亡比例明显增加（P＜0.01）；[Ca2＋]i显著降低（P＜0.05）。结论leptin‐siRNA可以通过沉默leptin基因表达而抑制 HSC的增殖、促进凋亡和减少[Ca2＋]i 。leptin在肝纤维化发展中起着重要作用，有望成为肝纤维化一个新的预测和预后的标志。
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