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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 investigate the effect of compound Biejia Ruangan prescription on the expression of nuclear factor-kappaB (NF-κB) and inhibitory protein family (IκB) in renal tubular epithelial cells at different time points. Methods UUO rats were used to prepare drug-containing serum. 5% drug-containing serum medium was added to cultured renal tubular epithelial cells. At different times, the expression of NF-κB and IκB in each group was detected by Western blot. RESULTS: LPS stimulated renal tubular epithelial cells. After 6 h, the activity of NF-κB in the cytoplasm gradually increased; nuclear transfer of NF-κB occurred in 12 h; the activation of NF-κB in the nucleus continued until 24 h. After 2 hours of LPS stimulation, the expression of IκBα in the cytoplasm was the lowest point, gradually recovered after 4 hours, and returned to normal level after 8 hours. Conclusion The compound BJJR serum can inhibit the activation of NF-κB and prevent the degradation of IκBα, so that NF-κB can not be dissociated from the NF-κBIκBα complex and block the nuclear transfer of NF-κB. It is one of the mechanisms for preventing and treating renal interstitial fibrosis.