Inhibition of Hepatic Stellate Cell Proliferation and Extracellular Ca2+ Influx by Leptin-siRNA
10.3870/j.issn.1672-0741.2014.06.004
- VernacularTitle:靶向瘦素基因小干扰RNA抑制肝星状细胞增殖和细胞内钙离子浓度
- Author:
Xiulan XUE
;
Jusheng LIN
- Publication Type:Journal Article
- Keywords:
leptin;
small interfering RNA;
apoptosis;
hepatic stellate cell;
intracellular Ca2+ concentration
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2014;(6):631-635
- CountryChina
- Language:Chinese
-
Abstract:
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.
