金钗石斛多糖对高糖诱导的大鼠肾系膜细胞Nrf2表达的影响

The effect of Dendrobium nobile polysaccharides on expression of Nrf2 in rat glomerular mesangial cell cultured in high glucose

目的观察金钗石斛多糖对高糖作用下大鼠肾小球系膜细胞(HBZY-1)Nrf2表达的影响。方法将HBZY-1细胞分为6组,对照组(NC)、葡萄糖作用组(HG)、NAC组(NA)、金钗石斛多糖低(LD)、中(MD)、高(HD)剂量干预组,药物作用时间24h,Western blot检测细胞内Nrf2蛋白表达,细胞免疫荧光检测FDP1对Nrf2核转位.结果与高糖作用组相比,金钗石斛多糖低中高剂量组均能明显上调Nrf2mRNA、Nrf2蛋白的表达,促进细胞Nrf2蛋白的核转位,上调了NQO1mRNA的表达(P<0.05)。结论金钗石斛多糖FDP1通过上调Nrf2mRNA、Nrf2蛋白表达和Nrf2蛋白核移位上调其调控的下游基因NQO1mRNA,以抵抗高糖诱导下氧化应激损伤,从而保护细胞的氧化还原稳态。

Objective To explore the effect of Dendrobium Mobile polysaccharides rat mesangial ceils induced by high glucose and possible mechanism of polysaccharides involved in enhancing cell expression of Nrf2. Methods The HBZY - 1 cells were divided into normal control group （NC） ,high glucose group （HG） ,the NAC treated group （NA） ,the Dendrobiurn Mobile polysaccharides low （LD）, medium （MD）, high （HD） dose group, drug action 24h, total antioxidant capacity of cells in each group were detected by ABTS method;reactive oxygen species by flow cytometry; cell proliferation in each group was examined by WST - 1 assay~ Nrf2mRNA, NQOlmRNA, NOS2mRNA expression level were detected by real- time fluorescence quantitative PCR and nuclear translocation of immunofluorescent cells was observed. Intracellular Nrf2 protein expression was detected by immunodetection. Re- suits Nrf2mRNA and Nrf2 protein expression levels were significantly increased in FDP1 treated groups （P 〈 0.05）, and the Nrf2 protein nuclear translocation was promote. Meanwhile, NQOImRNA and NOS2mRNA expression levels were also increased （P 〈 0.05 ）. Conclusion Through activation of the Nr^2 pathway, Dendrobium Mobile polysaccharide could enhance cell resistance to ox- idation, protect and maintain intracellular redox homeostasis.