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目的:观察筋骨草总黄酮(TFA)含药血清对肾小球系膜细胞(GMC)p38丝裂原激活蛋白激酶/核转录因子-κB(p38MAPK/NF-κB)信号通路的影响,进一步探讨TFA保护肾脏的可能机制。方法:体外培养大鼠GMC,以脂多糖(LPS)诱导大鼠GMC增殖,加入TFA含药血清进行干预。采用ELISA法检测细胞上清液中IL-1β、TNF-α的含量;RT-PCR法检测NF-κB mRNA和IκB m RNA的表达水平;Western Blot法检测p-p38MAPK蛋白的表达情况。结果:TFA含药血清干预24、48h后细胞上清液中IL-1β、TNF-α的含量较LPS组均明显减少(P<0.05,P<0.01)。干预48h后,IκB m RNA的相对表达量较LPS组明显增多,而NF-κB mRNA和p-p38MAPK蛋白的相对表达量均明显减少(P<0.05,P<0.01)。结论:TFA的肾脏保护作用可能与调控p38MAPK/NF-κB信号通路,继而减弱下游炎性因子IL-1β、TNF-α的表达有关。
OBJECTIVE: To investigate the effect of serum containing total flavonoids from Radix Astragali (TFA) on p38 mitogen-activated protein kinase / nuclear factor-κB (p38MAPK / NF-κB) signaling in glomerular mesangial cells (GMC) TFA protects the kidney possible mechanism. Methods: The rat GMCs were cultured in vitro. The proliferation of rat GMCs was induced by lipopolysaccharide (LPS). TFA-containing serum was added for intervention. The contents of IL-1β and TNF-α in the cell supernatant were detected by ELISA. The expression of NF-κB mRNA and IκB m RNA were detected by RT-PCR. The expression of p-p38MAPK protein was detected by Western Blot. Results: Compared with LPS group, the contents of IL-1β and TNF-α in supernatant of TFA-containing serum were significantly decreased at 24 h and 48 h (P <0.05, P <0.01). Compared with LPS group, the relative expression of IκB m RNA significantly increased (P <0.05, P <0.01), while the relative expression of NF-κB mRNA and p-p38MAPK protein decreased significantly after 48h intervention. CONCLUSION: The renal protective effect of TFA may be related to the regulation of p38MAPK / NF-κB signaling pathway and the subsequent down-regulation of the expression of downstream inflammatory cytokines IL-1β and TNF-α.