山茱萸、生地有效部位及配伍组合抑制糖基化终末产物致肾小球系膜细胞增殖的研究  

Effect of the effective components and combinations thereof in Corni Fructus and Radix Rehmanniae on glomerulus mesangial cell proliferation induced by AGEs

作　　者：吴云皓[1] 李伟[1] 陈玉萍[1] 周芷若[1] 许惠琴[1] 

机构地区：[1]南京中医药大学药学院,江苏省中药药效与安全性评价重点实验室,南京210023

出　　处：《中药药理与临床》2015年第4期71-75,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金资助项目(No.81073111,No.81374029);江苏高校优势学科建设工程资助项目(No.nzyzyxj P1006)

摘　　要：目的：通过体外培养大鼠肾小球系膜细胞（GMC）,建立糖基化终末产物（AGEs）致肾小球系膜细胞增殖的细胞模型,观察山茱萸有效部位、生地有效部位及其配伍组合对糖基化终末产物致肾小球系膜细胞增殖的抑制作用,以确定最佳配伍组合物。方法：体外培养肾系膜细胞,当细胞贴壁生长至80%-90%时,（1）,用浓度为50、100、200、400mg/L的山茱萸各部位（环烯醚萜苷、三萜酸、多糖）、生地各部位（环烯醚萜苷、多糖、寡糖）进行干预,然后加入200mg/L的糖基化终末产物刺激,24 h后检测细胞增殖情况,筛选出活性较强的部位。（2）,将活性较强的部位山茱萸环烯醚萜苷和三萜酸（A）、生地环烯醚萜苷（B）按1：0、0：1、1：1、1：2、2：1进行配伍组合（分别为A1、B1、A1B1、A1B2、A2B1）,以浓度为25、50、100mg/L生药及含药血清进行干预,加入200mg/L的AGEs刺激,24 h后检测细胞增殖情况以及细胞上清液中纤维连接蛋白（FN）、层粘连蛋白（LN）、Ⅳ型胶原（Coll-Ⅳ）水平。结果：（1）山茱萸环烯醚萜苷部位、山茱萸三萜酸部位与生地环烯醚萜苷部位活性较强,具有显著抑制糖基化终末产物致肾小球系膜细胞增殖的作用。（2）配伍组合物（A1、B1、A1B1、A1B2、A2B1）生药及含药血清均能够抑制肾小球系膜细胞增殖及其分泌纤维连接蛋白、层粘连蛋白、Ⅳ型胶原的水平,并呈一定量效关系。结论：山茱萸环烯醚萜苷部位、山茱萸三萜酸部位、生地环烯醚萜苷部位及其配伍组合能够明显抑制糖基化终末产物致肾系膜细胞增殖及其分泌的纤维连接蛋白、层粘连蛋白、Ⅳ型胶原水平,减少细胞外基质沉积,其中配伍组合A2B1抑制效应最强,为最佳配伍组合。Objective： To observe the inhibition of the effective components and combinations thereof in Cornus Fructus（ A） and Radix Rehmanniae（ B） on glomerulus mesangial cells（ GMC） which are induced by advanced glycation end products（ AGEs）,and screen the optimized combination. Methods： GMC were cultured in vitro,when the cell attachment rate is about 80% - 90%,（ 1） GMC were intervened with components in Corni Fructus（ iridoid glycosides,triterpenic acids,polysaccharides） and Radix Rehmanniae（ iridoid glycosides,polysaccharides,oligosaccharides）,then stimulated by 200 mg / L AGEs,the cell proliferation was tested by MTT,and the stronger active components were screened.（ 2） The crude drugs and drug-containing serum of the combinations（ A1,B1,A1B1,A1B2,A2B1） of the stronger active components including Corni Fructus iridoid glycosides,Corni Fructus triterpenic acids（ A） and Radix Rehmanniae iridoid glycosides（ B） in the ratio of 1： 0,0： 1,1： 1,1： 2,2： 1 were used to intervene the GMC induced by 200 mg / L AGEs. The cell proliferation and FN,LN,Coll-Ⅳ levels in the supernatant were tested by MTT assay after 24 hours. Results：（ 1） Corni Fructus iridoid glycosides,Corni Fructus triterpenic acids and Radix Rehmanniae iridoid glycosides had stronger effects against GMC proliferation.（ 2） The crude drugs and drug-containing serum of the combinations（ A1,B1,A1B1,A1B2,A2B1） inhibited GMC proliferation and reduced the levels of FN,LN,Coll-Ⅳ,and there was dose-effect relation. Conclusion： Corni Fructus iridoid glycosides,Corni Fructus triterpenic acids,Radix Rehmanniae iridoid glycosides and combinations thereof could inhibit GMC proliferation and reduce the levels of FN,LN,Coll-Ⅳ,decreasing the accumulation of extracellular matrix. The combination A2B1 which produced the strongest effect was thought to the best combination.

关 键 词：山茱萸 生地 有效部位 配伍组合 肾小球系膜细胞 

分 类 号：R285[医药卫生—中药学]