复方鳖甲软肝方对LPS刺激肾小管上皮细胞NF-κB、IκBα表达的影响

Influence of Fufangbiejiaruangan Formula on expressions of NF-κB and IκBαin TEC stimulated by LPS

目的研究复方鳖甲软肝方对肾小管上皮细胞不同时段表达核转录因子(NF-κB)、抑制蛋白家族(IκB)的影响。方法由UUO大鼠制备含药血清,将5%含药血清培养基加入体外培养的肾小管上皮细胞,在不同时间,采用免疫印迹法(Westernblot)检测各组NF-κB、IκB的表达。结果LPS刺激肾小管上皮细胞,6h后,细胞胞浆内NF-κB活性逐渐增强;12hNF-κB发生了核转移;细胞核内NF-κB的活化持续到24h后。LPS刺激2h后,细胞浆中IκBα的表达为最低点,4h后逐渐回升,8h后基本回复正常水平。结论复方鳖甲软肝方含药血清能抑制NF-κB的活化,并阻止IκBα的降解,使NF-κB无法从NF-κBIκBα复合物上解离下来,阻止了NF-κB的核转移,可能是其防治肾间质纤维化的机制之一。

Objective To study the influence of Fufangbiejiaruangan Formula on the expressions of NF- κB and IκBα in tubules epithelial cells （TEC） in different periods. Method The serum with Fufangbiejiaruangan Formula was prepared in rats with uniureteroligation （UUO） and then the culture with 5% serum was given to TEC in vitro. The Western blotting method was used to detect the expressions of NF-κB and IκBα at different time points. Result TEC was stimulated with LPS and 6 hours later the activity of NF-κB in eytoplasma was increased, and karyon shifted after 12 hours. The activation of NF-κB in karyon continued to 24th hour. TEC was stimulated with LPS and 2 hours later the expression of IκBα reached the lowest point, it increased after 4 hours and reached the normal level after 8 hours. Condution Fufangbiejiaruangan Formula can inhibit the activation of NF-κB and stop the degradation of IκBα, prevent the dissociation of NF-κB from the compound of NF-κB and IκBα, and stop the karyon shift of NF-κB. This may be the mechanism of Fufangbiejiaruangan Formula for preventing and treating renal interstitial fibrosis.