银杏叶提取物对大鼠肾小球系膜细胞中细胞外基质及β_1整合素表达的影响

Effect of Extract Ginkgo Biloba on Extracellular Matrix and β_1-Integrin Expressions of Glomerular Mesangial Cells in Rats

目的观察银杏叶提取物(EGB)对脂多糖(LPS)诱导的大鼠肾小球系膜细胞(GMC)中细胞外基质(ECM)分泌及β1整合素(Itgβ1)表达的影响。方法体外培养的大鼠GMC鉴定后第3～10代用于实验。实验分为5组:对照组,LPS组,EGB高、中、低剂量组。酶联免疫吸附试验法测定6 h、12 h和24 h细胞培养上清液中ECM层黏连蛋白、纤维连接蛋白和Ⅳ型胶原蛋白水平;荧光半定量-PCR法检测Itgβ1 mRNA表达的变化。结果 1.正常培养的大鼠GMC可分泌一定量的ECM,LPS组在各时间点分泌ECM均高于对照组(Pa<0.01),而EGB各剂量组分泌ECM均低于LPS组(Pa<0.01);2.正常培养的大鼠GMC可表达一定量Itgβ1mRNA,LPS组各时间点Itgβ1 mRNA表达量均高于对照组(Pa<0.01),EGB各剂量组Itgβ1 mRNA表达量均低于LPS组(Pa<0.01)。结论LPS可诱导GMC Itgβ1 mRNA表达及ECM分泌增加,EGB可抑制LPS诱导的GMC Itgβ1 mRNA表达及ECM分泌增加,EGB可通过抑制细胞因子及ECM分泌等分子生物学机制对肾脏起保护作用。

Objective To observe the effects of extract Ginkgo biloba（EGB） on β1-integrin（Itgβ1） expression and extracellular matrix（ECM）secretion of glomerular mesangial cells（GMC） in rats induced by lipopolysaccharide（LPS）. Methods The cultured GMCs of rats were established in vitro,3-10 offspring of rat cells were used in the experiment after identification.Cultured GMCs were divided into 5 groups：control group,LPS group,high,middle and low dose EGB groups.The changes in ECM including laminin（LN）,fibronectin（FN） and collagen type Ⅳ（ColⅣ） protein secretion were detected by the enzyme-linked immunosorbent-assay.The changes of Itgβ1 mRNA expression was detected by semi-quantitative real-time reverse transcription-polymerase chain reaction. Results 1.GMC could secrete some ECM protein in normal culture medium,and GMC secreted ECM protein was higher significantly in LPS group than that in control group（Pa〈0.01）,but significantly lower in all EGB groups than that in LPS group（Pa〈0.01）.2.GMC could express low level Itgβ1 mRNA in normal culture medium,Itgβ1 mRNA expression was higher significantly in LPS group than that in control group at all time points（Pa〈0.01）,but lower significantly in EGB group than that in LPS group（Pa〈0.01）. Conclusions LPS can induce the increase of ECM secretion and Itgβ1 mRNA expression;EGB can inhibit the ECM secrection and Itgβ1 mRNA expression induced by LPS in GMC as dose-dependent manner.Renoprotective effect of EGB may be involved in inhibiting the ECM secretion and cytokines expression.