中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
2期
132-135
,共4页
谢佩玉%松仓诚%藤井绩%赵基恩%筱原诚%张晓梅
謝珮玉%鬆倉誠%籐井績%趙基恩%篠原誠%張曉梅
사패옥%송창성%등정적%조기은%소원성%장효매
高糖%MIN-6细胞%视网膜色素上皮细胞
高糖%MIN-6細胞%視網膜色素上皮細胞
고당%MIN-6세포%시망막색소상피세포
High glucose%MIN-6 cell%Retinal pigment epithelial cell
背景 糖尿病视网膜病变(DR)与高糖所致的视网膜色素上皮(RPE)细胞的氧化损伤密切相关,近年来人们广泛地致力于保护RPE细胞的研究.目的 研究胰岛β细胞株MIN-6细胞对RPE细胞高糖损伤的保护作用. 方法 将RPE细胞体外培养4d,待贴壁生长良好后分为正常对照组、高糖对照组和MIN-6细胞共培养组,ABC法鉴定RPE细胞.用含5 mmol/L葡萄糖的RPE细胞培养液进行培养作为正常对照组,高糖对照组用含30 mmol/L葡萄糖的培养液进行培养,MIN-6细胞共培养组为含5×104个MIN-6细胞以及30 mmol/L葡萄糖的培养液进行培养.各组继续培养24 h后,应用四甲基偶氮唑蓝(MTT)法测量高糖对已贴壁生长良好的RPE细胞的损伤作用以及MIN-6细胞对RPE细胞高糖损伤的保护作用. 结果 ABC法鉴定RPE细胞可见95％以上呈棕色着色,为阳性细胞.各组继续培养24 h后,正常对照组、高糖对照组和MIN-6细胞共培养组RPE细胞的吸光度(A550)值差异有统计学意义(F=19.94,P＜0.01),正常对照组的A550值为0.44±0.02,高糖对照组为0.30±0.01,差异有统计学意义(t=6.85,P＜0.01)；MIN-6细胞共培养组的A550值为0.41±0.01,与高糖对照组的0.30±0.01比较,差异有统计学意义(t=5.62,P＜0.01).正常对照组RPE细胞的生存率为(97.5±3.3)％,高糖对照组为(68.2±4.5)％,差异有统计学意义(t=11.30,P＜0.01).结论 高糖导致贴壁生长良好的RPE细胞损伤,MIN-6细胞可以明显保护高糖所致的RPE细胞损伤.
揹景 糖尿病視網膜病變(DR)與高糖所緻的視網膜色素上皮(RPE)細胞的氧化損傷密切相關,近年來人們廣汎地緻力于保護RPE細胞的研究.目的 研究胰島β細胞株MIN-6細胞對RPE細胞高糖損傷的保護作用. 方法 將RPE細胞體外培養4d,待貼壁生長良好後分為正常對照組、高糖對照組和MIN-6細胞共培養組,ABC法鑒定RPE細胞.用含5 mmol/L葡萄糖的RPE細胞培養液進行培養作為正常對照組,高糖對照組用含30 mmol/L葡萄糖的培養液進行培養,MIN-6細胞共培養組為含5×104箇MIN-6細胞以及30 mmol/L葡萄糖的培養液進行培養.各組繼續培養24 h後,應用四甲基偶氮唑藍(MTT)法測量高糖對已貼壁生長良好的RPE細胞的損傷作用以及MIN-6細胞對RPE細胞高糖損傷的保護作用. 結果 ABC法鑒定RPE細胞可見95％以上呈棕色著色,為暘性細胞.各組繼續培養24 h後,正常對照組、高糖對照組和MIN-6細胞共培養組RPE細胞的吸光度(A550)值差異有統計學意義(F=19.94,P＜0.01),正常對照組的A550值為0.44±0.02,高糖對照組為0.30±0.01,差異有統計學意義(t=6.85,P＜0.01)；MIN-6細胞共培養組的A550值為0.41±0.01,與高糖對照組的0.30±0.01比較,差異有統計學意義(t=5.62,P＜0.01).正常對照組RPE細胞的生存率為(97.5±3.3)％,高糖對照組為(68.2±4.5)％,差異有統計學意義(t=11.30,P＜0.01).結論 高糖導緻貼壁生長良好的RPE細胞損傷,MIN-6細胞可以明顯保護高糖所緻的RPE細胞損傷.
배경 당뇨병시망막병변(DR)여고당소치적시망막색소상피(RPE)세포적양화손상밀절상관,근년래인문엄범지치력우보호RPE세포적연구.목적 연구이도β세포주MIN-6세포대RPE세포고당손상적보호작용. 방법 장RPE세포체외배양4d,대첩벽생장량호후분위정상대조조、고당대조조화MIN-6세포공배양조,ABC법감정RPE세포.용함5 mmol/L포도당적RPE세포배양액진행배양작위정상대조조,고당대조조용함30 mmol/L포도당적배양액진행배양,MIN-6세포공배양조위함5×104개MIN-6세포이급30 mmol/L포도당적배양액진행배양.각조계속배양24 h후,응용사갑기우담서람(MTT)법측량고당대이첩벽생장량호적RPE세포적손상작용이급MIN-6세포대RPE세포고당손상적보호작용. 결과 ABC법감정RPE세포가견95％이상정종색착색,위양성세포.각조계속배양24 h후,정상대조조、고당대조조화MIN-6세포공배양조RPE세포적흡광도(A550)치차이유통계학의의(F=19.94,P＜0.01),정상대조조적A550치위0.44±0.02,고당대조조위0.30±0.01,차이유통계학의의(t=6.85,P＜0.01)；MIN-6세포공배양조적A550치위0.41±0.01,여고당대조조적0.30±0.01비교,차이유통계학의의(t=5.62,P＜0.01).정상대조조RPE세포적생존솔위(97.5±3.3)％,고당대조조위(68.2±4.5)％,차이유통계학의의(t=11.30,P＜0.01).결론 고당도치첩벽생장량호적RPE세포손상,MIN-6세포가이명현보호고당소치적RPE세포손상.
Background Diabetic retinopathy (DR) associated with this disease closely approximates the oxidative damage inflicted on retinal pigment epithelial (RPE) cells by high glucose,in recent years,people devote themselves to the protection of RPE cells extensively. Objective To investigate the protective effect of pancreatic β MIN-6 cells on retinal pigment epithelial(RPE) cells from high glucose-induced damage. Methods RPE cells were incubated with normal medium for 4 d and divided into 3 groups:normal glucose group,high glucose group and MIN-6 cells group.RPE cell were exposed with 5 mmol/L normal glucose in normal glucose group,exposed with 30 mmol/L high glucose in high glucose group,and exposed with 5 × 104 MIN-6 cells and 30 mmol/L high glucose in MIN-6 cells group.After 24 h,cell viability of RPE cells was determined by MTT cell viability assay. Results More than 95％ cells showed the brown staining by ABC method.After incubation for another 24 hours,the A550 value was 0.44±0.02,0.30±0.01 and 0.41±0.01 in the normal glucose group,high glucose group and MIN-6 cells group respectively with a significant difference among these three groups (F =19.94,P＜ 0.01 ).The A55o value was significantly higher in the normal glucose group and the MIN-6 cells group compared with the high glaucose group (t =6.85,5.62,P＜0.01 ).The survival rate of RPE cells in the normal glucose group was(97.5±3.3 )％,and that in the high glucose group was ( 68.2 ± 4.5 ) ％,showing significant difference between them ( t =11.30,P＜0.01 ).Conclusions High glucose-induced damage of RPE cells is abrogated,and MIN-6 cells can protect RPE cells from high glucose-induced damage.
