补肾活血中药血清对高糖条件下纯化培养的视网膜神经节细胞Glu释放量的影响

Effect of Drug-Serum with Chinese Drugs for Nourishing Shen and Activating Blood on the Glutamate Release in Purified Retinal Ganglion Cells Cultured in High-glucose Conditions

免费下载全文查看/发表评论下载PDF阅读器

DOI：

中文关键词: 高糖糖波动视网膜神经节细胞谷氨酸补肾活血

英文关键词:high-glucose fluctuated glucose condition retinal ganglion cells glutamate nourishing Shen and activating blood

基金项目:国家自然科学基金资助项目(No.30572407);四川省科技厅应用基础项目(No.008JY0085);四川省教育厅重点项目(No.07EA023)

作者	单位
马荣	成都中医药大学研究生院
谢学军	成都中医药大学眼科教研室
万李	成都中医药大学研究生院
马殿伟	成都中医药大学研究生院
米珍	成都中医药大学研究生院

摘要点击次数: 1425

全文下载次数: 775

中文摘要:

目的探讨稳定高糖及糖波动条件对体外纯化培养的视网膜神经节细胞(retinal ganglion cells,RGCs)谷氨酸(glutamate,Glu)释放量的影响,以及补肾活血中药的干预作用。方法采用抗体联合两步纯化法培养新生Sprague-Dawley(SD)大鼠RGCs,将其分别置于模拟正常条件、稳定高糖条件及糖波动条件下进行培养,并以补肾活血中药复方含药血清进行干预。实验分为正常对照组、正常中药干预组,稳定高糖组、稳定高糖中药干预组、糖波动组、糖波动中药干预组;分别在试验干预24、48、72h后,用全自动氨基酸分析仪测定各组细胞外液中Glu的含量(mg/L),采用SPSS13.0软件对所得数据进行统计学分析。结果糖波动组24h时RGCs的Glu释放量[(256.33±25.73)mg/L]较同期正常对照组[(134.22±9.14)mg/L]及稳定高糖组[(141.17±22.13)mg/L]均明显增加(P<0.05)。正常中药干预组的Glu释放量在24h[(124.50±10.30)mg/L]及72h[(30.17±2.97)mg/L]时段均较正常对照组降低(P<0.05);稳定高糖中药干预组24h[(127.50±16.94)mg/L]、48h[(26.17±3.99)mg/L]及72h[(27.67±3.49)mg/L]时段的Glu释放量均较稳定高糖组降低(P<0.05);糖波动中药干预组RGCs的Glu释放量在24h[(228.33±18.41)mg/L]、72h[(28.00±2.41)mg/L]均较糖波动组降低(P<0.05)。结论糖波动条件能明显增加RGCs的Glu释放量,导致大量Glu在细胞外聚积,细胞外高浓度Glu聚积最终引发RGCs神经兴奋性毒性,加剧细胞损害;补肾活血中药能在一定程度降低稳定高糖及糖波动条件下RGCs的Glu释放量,减轻Glu神经兴奋性毒性作用对RGCs的损伤,这可能是补肾活血中药防治糖尿病性视网膜病变(diabetic retinopathy,DRP)的药物干预途径之一。

英文摘要:

Objective To investigate the impacts of steady high-glucose or fluctuated glucose conditions on glutamate(Glu) release in purified retinal ganglion cells(RGCs) cultured in vitro,and the effect of serum contained Chinese drugs for nourishing Shen and activating blood(S-NSAB) on it.Methods RGCs of neonatal SD rats were cultured by antibody combined two-step purified method in different conditions:the simulated normal condition,the steady high-glucose condition and the fluctuated glucose condition,and they were intervened with S-NSAB.Thereby,the experiment was carried out in 6 groups,i.e.the normal control group(A),the S-NSAB intervened group(B),the steady high-glucose cultured group(C),the steady high-glucose cultured and SNSAB intervened group(D),the fluctuated glucose cultured group(E),and the fluctuated glucose cultured and S-NSAB intervened group(F) .Content of Glu in the extracellular fluid was detected at 24,48 and 72 h after intervention with a full-automatic biochemical analyzer.And the data obtained were statistically analyzed with SPSS 13.0 soft ware.Results Release of Glu at 24 h after intervention in Group E(256.33 ±25.73 mg/L) was obviously higher than that in Group A and Group C(134.22 ±9.14 mg/L and 141.17 ±22.13 mg/L,P <0.05);at 24 h and 72 h in Group B(124.50 ±10.30 mg/L and 30.17 ±2.97 mg/L) was obviously lower than in Group Arespectively(P < 0.05);in Group D at 24 h(127.50 ± 16.94 mg/L),48 h(26.17 ± 3.99 mg/L) and 72 h(27.67 ±3.49 mg/L) were lower than in Group C;in Group F at 24 h(228.33 ±18.41 mg/L) and 72 h(28.00 ±2.41 mg/L) were lower than in Group E respectively at the corresponding time points.Conclusions Fluctuated glucose condition could obviously increase the Glu release of RGCs,to cause extracellular large amount Glu accumulation,which induces the exciting neurotoxicity to RGCs and finally to aggravate the injury on cells.S-NSAB could reduce the Glu release to some extent in the steady-high or fluctuated glucose conditions,diminish the injury of RGCs from exciting neurotoxicity of Glu,and it might be one of the intervening pathways of Chinese drugs for NSAB in preventing and treating DRP.

关闭