转化生长因子-β_1抗体对肝星状细胞内钙离子浓度的影响

Effect of transforming growth factor-β_1 antibody on intrace llular calcium ion in cultured hepatic stellate cells

目的: 研究转化生长因子-β1 (TGF-β1 )抗体对肝星状细胞内钙离子浓度([Ca2+]i)的影响,探讨其预防、治疗肝纤维化和门脉高压的分子机制. 方法: 分别用TGF-β1 抗体和四氯化碳(CCl4 )处理大鼠肝星状细胞,作用24 h后,以Fluo-3/AM为细胞内钙离子荧光指示剂负载细胞,通过激光共聚焦显微镜观察[Ca2+]i的变化. 再利用正交设计方法,研究不同浓度CCl4 、 TGF-β1 抗体及其作用的先后顺序和二者作用的时间间隔对[Ca2+]i 的影响. 结果: 不同浓度TGF-β1 抗体、CCl4 及其作用的前后顺序对[Ca2+]i有显著性影响(P<0.05),两因素作用的时间间隔对[Ca2+]i的影响无显著性(P>0.05). CCl4在5 ～15 mmol/L范围内显著增加[Ca2+]i (P<0.05),5 ～20 mg/L TGF-β1 抗体可降低[Ca2+]i (P<0.05). 结论: TGF-β1抗体可以缓解肝星状细胞内钙超载.

AIM： To study the effect of transforming growth factor-β1, （TGF-β1,） antibody on intracellular calcium ion （ [ Ca^2＋ ]i） in hepatic stellate cells （HSC） and to explore the molecular mechanism in prevention and treatment of hepatic fibrosis and portal hypertension. METHODS： The activated HSC line was plated on small glass coverslips in ：24well culture dishes at a density of 5 × 10^9/L and stimulated by carbon tetrachloride （ CCl4 ） and TGF-β1, antibody for 24 h respectively. After the cells were loaded with Fluo-3/AM, [ Ca^2＋ ]i was measured by laser scanning confocal microscopy （LSCM）. Orthogonal design was used to study the changes of [Ca^2＋ ]i in HSCs treated with different concentrations of CCl4 and TGF-β1, antibody in different order and at different interval time. RESULTS： The [ Ca^2＋ ]i was significantly different from different concentrations of TGF-β1 and CCl4 and different orders of treatment （ P 〈 0. 05 ）, while no significance was found between different interval times （P〉0.05）. CCl4 sharply increased [Ca^2＋ ]i when the dosage ranged from 5 to 15 mmol/L （ P〈0.05）, while 5 to 20 mg/L TGF-β1, antibody decreased [ Ca^2 ＋ ]i （ P 〈 0.05 ）. CONCLUSION： TGF-β1, antibody can remit the [Ca^2＋ ]i overloading in HSC.