多球壳菌素对高糖诱导肾小球系膜细胞肥大及细胞外基质合成的影响

Effect of Myriocin on Hypertrophy of Glomerular Mesangial Cells and Extracellular Matrix Production Induced by High Glucose

目的 观察高糖对大鼠肾小球系膜细胞（GMC）大小及分泌细胞外基质（ECM）成分的影响及从冬虫夏草中提取的免疫抑制活性成分——多球壳菌素（ISP-1）的干预效应。方法正常糖（5．6mmol／LD-glucose）及高糖（25.2mmol／LD-glucose）条件下培养GMC，并设高糖加ISF1（100mg／L）组。采用流式细胞仪测各组GMC体积[前向角散射光（FSC）平均强度表示细胞大小。ELISA检测高糖条件下不同时间点（0、24、48、72h）细胞培养上清液中纤维连接蛋白（FN）、Ⅳ型胶原（ColⅣ）、Ⅲ型前胶原（PoolⅢ）、透明质酸（HA）及层黏连蛋白（LN）含量及ISP-1干预后分泌水平变化。结果同正常糖组比较。高糖可使GMC发生肥大，同时高糖环境下GMC分泌的Col、ColⅣ、FN、PcolⅢ、HA及LN明显增加（P〈0．05），48-72h更明显；ISPd能有效抑制高糖的上述作用（P〈0．05）。结论高糖可致GMC肥大和ECM合成增加,ISP-1对高糖所致GMC肥大和ECM合成增加有抑制作用，有望用于糖尿病肾病防治。

Objective To observe the changes of extracellular matrix （ECM） proluetion and hypertrophy of glomerular mesangial cells （GMCs） induced by high glucose（HG）, and the inhibitory role of myriocin（ 1SP-1）. Methods GMC cultured with norranl glucose （5.6 mmol/L D-Glucose, NG）, HG （25.2 mmol/L D-Glucccse） and H（7, plus ISP-1（ 100 mg/L） for different durations （ 0, 24,48, 72 h）. The sizes of GMC were indicated by forward scatter intensity,measured by flow cytometery,and the levels of fibronection（FN）, collagen Ⅳ （Col Ⅳ ） ,laminin（LN） ,precollagen Ⅲ （Pool Ⅲ ） and hyaluronic acid（HA） in the supematant of cultured GMC were detected by ELISA, Results Compared with NG, HG could induce GMC hypertrophy（ P 〈 0.05 ）, the levels of FN, Col Ⅳ, LN, Pcol Ⅲ and HA in the supematant of cultured GMC were higher in PIG group than those in NG group（ P 〈 0.05 ）. Compared with HG, HG plus ISP-1 significantly attenuated the GMC hypertrophy（ P 〈 0.05 ）, and lowered ECM production of cultured GMC（ P 〈 0.05 ）, and it was more obvious during 48 to 72 hour. Conclusions High concentration of extracellular glucose can induce hypertrophy and ECM synthesis of GMC, which are significantly inhibited by ISP-1. Therefore, ISP-1 may be used in the prevention and treatment of diabetic nephropathy.