摘要
目的探讨蛋白磷酸酶2A(PP2A)的表达在肝枯否细胞(KC)核因子-κB(NF-κB)活化中的作用,对KC核因子-κB活化的表达调控机制进行理论探索。方法采用胶原酶灌注和percol密度梯度离心法分离得到高纯度大鼠KC,分为内毒素(LPS)及二硫代氨基甲酸吡咯烷(PDTC)+LPS分别与KC体外共培养组及一个阴性对照组,24 h后收集各组细胞和培养上清,分别采用RT-PCR法和酶联免疫吸附法(ELISA)检测KC中PP2A、NF-κB p65及TNF-α的表达和培养上清中TNF-α的含量。结果 LPS组细胞培养上清中TNF-α含量显著高于对照组(P<0.01),KC中NF-κB p65和TNF-α的表达也明显增强(P<0.01),但对照组的PP2A表达较LPS组强(P<0.01);PDTC+LPS组KC中NF-κB p65和TNF-α的表达较LPS组明显减弱(P<0.01),但PDTC+LPS组PP2A的表达较LPS组有明显增强(P<0.05)。结论 LPS能够显著增强KC中NF-κB p65的表达进而增强其TNF-α的表达,这种增强作用可能与PP2A的表达被抑制有关;用PDTC抑制NF-κB活性能够降低KC中TNF-a的表达,这种抑制作用可能与PP2A的表达增强有关;PP2A可能参与了肝枯否细胞核因子-κB活化的表达调控。
Objective To investigate the potential role of protein phosphatase 2A（PP2A） in nuclear factor-κB（NF-κB） activation of kupffer cells and its effective mechanism.Methods KCs of hyper purity were isolated from livers of healthy male adult rats and cultured.The KCs were divided into three groups,one group co-cultured with lipopolysaccharide（LPS）,and another one with Pyrrolidine dithiocarbamate（PDTC） and LPS,and the negative control group.After twenty-four hours,the cells and cultured supernatant were collected,the expression changes of NF-κB,PP2A and tumor necrosis factor-α（TNF-α） in kupffer cells were detected by RT-PCR and the TNF-α in cultured supernatants were by enzyme-linked immunosorbent assay.Results The level of TNF-α in cultured supernatants of LPS group was higher than the contral group（P0.01）,and the expression of NF-κB and TNF-α in KCs of LPS group were higher too（P0.01）,but the expression of PP2A was higher in the contral group（P0.01）;Contrast to LPS group,the expression of NF-κB and TNF-α in PDTC group were went weaker（P0.01）,but the expression of PP2A enhanced obviously（P0.05）.Conclusions LPS could result in NF-κB activation and NF-κB associated up regulation of TNF-α expression in Kupffer cells,and this enhancement property maybe relate to the suppressed expression of PP2A.And PDTC,the inhibitor of NF-κB can depress the expression of TNF-α in Kupffer cells which stimulated by LPS,and this depressant effect may have something to do with the up-regulation of PP2A.PP2A may play a regulatory role in NF-κB activation of kupffer cells.
出处
《肝胆外科杂志》
2011年第6期467-470,共4页
Journal of Hepatobiliary Surgery
