野生型PTEN过表达对体外活化肝星状细胞内钙离子浓度的影响

发布时间：2018-03-19 22:14

本文选题：肝星状细胞　切入点：第号染色体缺失的磷酸酶张力蛋白同源物基因　出处：《中国现代医学杂志》2017年28期 　论文类型：期刊论文

【摘要】：目的探讨野生型第10号染色体缺失的磷酸酶张力蛋白同源物基因(PTEN)过表达对体外活化肝星状细胞(HSC)内钙离子浓度的影响。方法体外培养活化大鼠肝星状细胞系(HSC-T6),利用腺病毒载体,将野生型PTEN基因转染活化HSC;Western blot及实时荧光定量聚合酶链反应(q RT-PCR)检测HSC的PTEN蛋白及m RNA表达;采用钙离子荧光探针Rhod-2/AM,于激光扫描共聚焦显微镜下检测HSC内钙离子浓度变化。实验分组:(1)Control组:腺病毒转染时以DMEM代替病毒液;(2)Ad-GFP组:转染表达绿色荧光蛋白(GFP)的对照空病毒Ad-GFP;(3)Ad-PTEN组:转染携带野生型PTEN基因并表达GFP的重组腺病毒Ad-PTEN。结果野生型PTEN在活化大鼠HSC大量表达,3组HSC内钙离子浓度比较,差异有统计学意义(P0.05),Ad-PTEN组HSC内钙离子浓度(251.60±90.88)低于Control组(1 953.95±132.99)及Ad-GFP组(1 937.57±115.17),而Control组与Ad-GFP组之间HSC内钙离子浓度比较,差异无统计学意义(P0.05)。结论野生型PTEN过表达可降低体外活化大鼠HSC内钙离子浓度。
[Abstract]:Objective to investigate the effect of overexpression of phosphatase tension protein congener (PTEN) on the concentration of calcium in activated hepatic stellate cells (HSCS) in vitro. Methods the activated rat hepatic stellate cell line HSC-T6 was cultured in vitro. Using adenovirus vectors, The expression of PTEN protein and m RNA in HSC were detected by transfection of wild-type PTEN gene into activated HSCC blot and real-time fluorescence quantitative polymerase chain reaction (Q RT-PCR). Calcium fluorescence probe Rhod-2 / AM was used to detect the change of calcium concentration in HSC under laser scanning confocal microscope. Ad-PTEN group: Ad-PTEN, a recombinant adenovirus carrying wild type PTEN gene and expressing GFP, was transfected into Ad-PTEN group. Results the Ca ~ (2 +) concentration of wild type PTEN in three groups of activated rat HSC was compared. There was a significant difference in the concentration of Ca ~ (2 +) in HSC between Ad-PTEN group and Control group (251.60 卤90.88) and in Ad-GFP group (1 937.57 卤115.17), respectively, and there was a significant difference between Control group and Ad-GFP group. Conclusion the overexpression of wild type PTEN can reduce the calcium concentration in HSC of rats activated in vitro.
【作者单位】：华北理工大学附属医院消化内科;
【基金】：河北省自然科学基金(No:H2013209327) 中国肝炎防治基金会天晴肝病研究基金资助项目(No:CFHPC20132078)
【分类号】：R575

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