枸杞多糖对高糖环境下肾小球系膜细胞基质的影响

EFFECTS OF LYCIUM BARBARUM POLYSACCHARIDES ON THE FORMATION OF EXTRACELLULAR MATRIX IN GLOMERULAR MESANGIAL CELLS CULTURED IN HIGH GLUCOSE

目的研究枸杞多糖对高糖环境下肾小球系膜细胞基质的影响。方法将细胞分为空白、正常糖、高糖及不同浓度(200、400和800mg/L)枸杞杞多糖干预共6组,MTT法检测细胞增殖情况,H2DCF-DA法检测细胞内活性氧水平,ELISA法检测细胞蛋白激酶C(PKC)活性,免疫细胞化学法检测转化生长因子β1(TGF-β1)及纤维连接蛋白(FN)的表达情况。结果高糖组分别与空白组、正常糖组比,其细胞内活性氧的生成和蛋白激酶C活性均显著增加(P<0.05)。枸杞多糖干预组可降低高糖刺激引起的氧化应激水平,抑制细胞过度增殖,同时降低PKC活性(P<0.05),能明显下调TGF-β1和FN的表达(P<0.05)。结论枸杞多糖通过抑制细胞增殖、减少细胞活性氧的生成、抑制蛋白激酶PKC活性、下调TGF-β1和FN的表达,从而抵抗高糖引起的系膜细胞的损伤,达到对肾小球系膜细胞的保护作用。

Objective To study the effects ofLycium barOarum polysacctianaes （Lm） on me iormauon of extrallu- lar matrix of human mesangial cells in high glucose. Methods Human mesangial cells were cultured in vitro and divided into control group, normal glucose group, high glucose group, and LBP groups at different concentrations （200, 400, 800 mg/L）, for 48 h. Endogenous ROS was measured by a fluorometric assay with 2＇,7＇-dichlorofluorescin diacetate （DCFH-DA）. The proliferation of mesangial cells was measured by MTT colorimetric method. The activity of protein kinase C （PKC） was measured by enzyme-linked immunosorbent assay （ELISA）. The expression of transforming growth factor-beta 1 （TGF-β1）, fibroneetin （FN） were detected by immunocytoehemistry. Results High glucose could promote the growth of cells, increase ROS production, PKC activity and decrease the expression of TGF-β1 and FN. LBP could inhibit the growth of cells, reduce the expression of TGF-β1, FN, PKC activity and ROS production in mesangial cells cultured under high glucose. Conclusion LBP could protect against the cellular damage induced by high glucose.