摘要
目的探讨罗格列酮(rosiglitazone,RSG)对高糖诱导视网膜神经节细胞(retinal ganglion cell,RGC)损伤的保护作用。方法体外培养大鼠RGC细胞株RGC-5细胞,50mol·L-1葡萄糖孵育细胞诱导损伤。应用CCK-8法测定并计算细胞的生长抑制率,流式细胞仪检测细胞凋亡,全自动氨基酸分析仪测定细胞谷氨酸(glutamic acid,Glu)释放量,测定细胞中丙二醛(malondialdehyde,MDA)含量以及超氧化物歧化酶(superoxide dismutase,SOD)活性。结果高糖(50mol·L-1)以时间依赖的方式抑制了RGC-5细胞的生长,高糖处理24h、48h和72h生长抑制率分别为(22.37±3.49)%、(42.18±6.34)%和(57.33±5.39)%(均为P<0.05);与高糖组比较,高糖+不同浓度RSG组(0.1×10-6mol·L-1、10-6mol·L-1、10×10-6mol·L-1)处理48h以剂量依赖的方式降低了高糖诱导的RGC-5细胞生长:生长抑制率高糖组(42.18±6.34)%,高糖+不同浓度RSG组分别为(35.66±4.73)%、(27.35±4.15)%和(25.17±3.42)%(均为P<0.05)。与对照组比较,高糖处理24h、48h和72h以时间依赖的方式促进了细胞凋亡(均为P<0.05);与高糖组比较,高糖+不同浓度RSG组(0.1×10-6mol·L-1、10-6mol·L-1和10×10-6mol·L-1)处理48h以剂量依赖的方式降低了高糖诱导的RGC-5细胞凋亡:高糖组凋亡率为(31.55±5.34)%,高糖+不同浓度RSG组分别为(23.75±3.72)%、(18.75±2.17)%和(17.53±3.05)%(均为P<0.05)。与对照组比较,高糖组Glu释放量显著增加:2组分别为(85.64±12.75)μg·L-1和(246.84±33.48)μg·L-1(P<0.05)。与高糖组比较,高糖+不同浓度RSG(0.1×10-6mol·L-1、10-6mol·L-1和10×10-6mol·L-1)处理48h组Glu的释放以剂量依赖的方式减少:高糖组Glu释放量为(246.84±33.48)μg·L-1,高糖+不同浓度RSG组分别为(175.34±23.69)μg·L-1、(117.25±18.76)μg·L-1和(109.34±15.54)μg·L-1(均为P<0.05)。与对照组比较,高糖组SOD活性显著降低:分别为(3.06±0.38)kU·g-1和(0.56±0.07)kU·g-1(均为P<0.05),而MAD水平显著增加:分别为(5.67±0.76)μmol·g-1和(37.64±4.37)μmol·g-1(均为P<0.05)。与高糖组相比较,高糖+不同浓度RSG处理48h组细胞中SOD活性以剂量依赖的方式增加(均为P<0.05),而MAD水平显著降低(均为P<0.05)。结论 RSG抑制了高糖诱导的RGC损伤,其机制与RSG减少了RGC中Glu的释放和抑制氧化应激有关。
Objective To study the protective effects of rosiglitazone on retinal ganglion cells （RGC） injury induced by high glucose.Methods Rats RGC-5 line cells were cultured in vitro,and incubated with 50 mol·L-1 glucose to induce the injury.The growth inhibitive rate was measured by CCK-8 analysis.The apoptotic rate was tested by flow cytometry.Content of glutamic acid in the medium was detected by a full-automatic biochemical analyzer.The content of malonaldehyde （MDA） and activity of superoxide dismutase （SOD） in cells were measured.Results High glucose （50 mmol·L-1） inhibited significantly the growth of RGC-5 cells in a time-dependent manner,the growth inhibitive rate were （22.37±3.49）%,（42.18±6.34）% and （57.33±5.39）% at 24 hours,48 hours and 72 hours after treatment,respectively（all P0.05）.Compared with high glucose group,the growth inhibitive rate of RGC-5 cells decreased in a dose-dependent manner in high glucose combined with rosiglitazone （0.1×10-6 mol·L-1,10-6 mol·L-1 and 10×10-6 mol·L-1） group for 48 hours,which were （42.18±6.34）%,（35.66±4.73）%,（27.35±4.15）% and （25.17±3.42）%,respectively（all P0.05）.Compared with control group,the apoptotic rate of RGC-5 cells in high glucose group increased significantly in a time-dependent manner （all P0.05）.Compared with high glucose group,apoptotic rate of high glucose combined with rosiglitazone （0.1×10-6 mol·L-1,10-6 mol·L-1 and 10×10-6 mol·L-1） group for 48 hours decreased in a dose-dependent manner,which were （31.55±5.34）%,（23.75±3.72）%,（18.75±2.17）% and （17.53±3.05）%,respectively （all P0.05）.Compared with control group,the release of Glu in RGC-5 cells significantly increased in high glucose group,which were （85.64±12.75）μg·L-1 and （246.84±33.48）μg·L-1,respectively（P0.05）.Compared with high glucose group,the release of Glu in RGC-5 cells of high glucose combined with rosiglitazone （0.1×10-6 mol·L-1,10-6 mol·L-1 and 10×10-6 mol·L-1） group for 48 hours decreased in dose-dependent manner,which were （246.84±33.48）μg·L-1,（175.34±23.69）μg·L-1,（117.25±18.76）μg·L-1 and （109.34±15.54）μg·L-1,respectively （all P0.05）.Compared with control group,SOD activity in high glucose group were significantly depressed,which were （3.06±0.38）kU·g-1 and （0.56±0.07）kU·g-1,respectively（P0.05） and the level of MDA greatly increased,which were （5.67±0.76）μmol·g-1 and （37.64±4.37）μmol·g-1,respectively （P0.05）.Compared with high glucose group,SOD activity in high glucose ＋ rosiglitazone（0.1×10-6 mol·L-1,10-6 mol·L-1 and 10×10-6 mol·L-1） group for 48 hours were significantly enhanced （all P0.05）,and the level of MDA （0.1×10-6 mol·L-1,10-6 mol·L-1 and 10×10-6 mol·L-1） group for 48 hours significantly decreased （all P0.05）.Conclusion Rosiglitazone can inhibit the high glucose-induced RGC injury in rat RGC-5 line cells,and the mechanism is related with the inhibition of glutamic acid release and oxidative stress induced by rosiglitazone.
出处
《眼科新进展》
CAS
北大核心
2013年第2期101-105,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:81100648)
衡阳市科学技术局科学技术计划项目(编号:2012KJ48)~~
关键词
罗格列酮
高糖
视网膜神经节细胞
谷氨酸
氧化应激
rosiglitazone
high glucose
retinal ganglion cells
glutamic acid
oxidative stress
