温脾汤药物血清对体外培养的大鼠系膜细胞核转录因子-κB活化的影响

INHIBITORY EFFECTS OF WENPITANG PHARMACOLOGIC SERUM ON ACTIVATION OF NF-κB IN MESANGIAL CELLS OF RATS in vitro

目的 探讨温脾汤药物血清对大鼠原代培养系膜细胞核转录因子NF κB及其抑制因子IκB表达的影响 ,提供其肾保护作用机理的实验依据。 方法 应用荧光标记和激光共聚焦扫描显微镜技术 ,研究温脾汤药物血清对LPS刺激引起的NF κB活化的影响 ;应用Westernblot方法研究温脾汤药物血清对LPS刺激引起的IκBα降解的影响。 结果 LPS刺激 10h后 ,NF κB的活性达到最高峰 ,且最高荧光强度位于细胞核内 ;大剂量药物血清明显抑制NF κB的活性增强 ;IκBα在LPS刺激后迅速降解 ,又在 1～ 2h内迅速恢复近正常水平 ;大剂量药物血清能明显抑制IκBα的降解。 结论 温脾汤通过抑制LPS诱导的IκBα降解 ,使NF κB与IκBα结合成复合物而减少解离 ,阻止了系膜细胞NF

Objective To observe the effect of Wenpitang pharmacologic serum on activation of NF\|κB and the degradation of its inhibitor IκBα in mesangial cells of rats in vitro. Provide experimental evidence for further evaluation of the renal protective effect of Wenpitang. Methods Effect of Wenpitang pharmacologic serum on activation of NF\|κB in the cells was investigated by using fluorescence\|labelling and laser confocal scanning microscopy;Effect of Wenpitang pharmacologic serum on degradation of IκBα was investigated by Western blot. Results Activation of NF\|κB in rat mesangial cells in vitro was maximal at 10 hours after stimulation by addition of LPS into culture medium; Degradation of the inhibitory subunit IκBα was observed soon after stimulation and had returned to near normal levels in 1\|2 hours.Treatment of mesangial cells with Wenpitang pharmacological serum in culture inhibited degradation of IκBα and prevented the activation of NF\|κB.Conclusion\ Wenpitang can partially inhibit the activation of NF\|κB in mesangial cells through inhibiting degradation of IκBα stimulated by LPS,which cause NF\|κB can not dissociate from NF\|κB/IκBα complex.\;[