中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2012年
9期
791-794
,共4页
视网膜%Müller细胞%葡萄糖%细胞培养%增生%凋亡
視網膜%Müller細胞%葡萄糖%細胞培養%增生%凋亡
시망막%Müller세포%포도당%세포배양%증생%조망
Retina%Müller cell%Glucose%Cell culture%Proliferation%Apoptosis
背景 视网膜Müller细胞具有为视网膜组织提供营养、维持视网膜的正常结构等多种生理功能,研究发现Müller细胞的病变会导致视网膜血管发生相应的改变.探讨高糖对视网膜Müller细胞的影响对于糖尿病视网膜病变(DR)的发病机制研究具有重要意义. 目的 研究不同浓度的葡萄糖对体外培养的视网膜Müller细胞活性的影响. 方法 取出生后10d清洁级SD大鼠的视网膜组织,用组织块培养法在含质量分数20％胎牛血清的DMEM培养液中体外原代培养Müller细胞并传代,取第3代细胞用免疫组织化学法对细胞进行鉴定.将不同浓度(5.5、30.0、40.0 mmol/L)的葡萄糖加入培养基中培养4d,MTT比色法测定各组波长570 nm处Müller细胞的吸光度(A570)值,计算各组细胞的相对存活率；采用流式细胞仪检测各组Müller细胞的凋亡率. 结果 培养的细胞贴壁生长,呈长梭形；95％以上细胞神经胶质纤维酸性蛋白(GFAP)反应阳性.MTT比色法检测显示,正常葡萄糖组及30.0 mmo/L、40.0 mmol/L葡萄糖处理组Müller细胞4570值分别为0.24±0.01、0.21±0.03和0.20±0.02,总体差异有统计学意义(F=6.755,P＜0.05).与正常葡萄糖组比较,30.0 mmol/L 、40.0 mmol/L葡萄糖处理组A570值均明显降低,差异有统计学意义(q=0.645、0.486,P＜0.05).流式细胞仪检测结果表明,正常葡萄糖组、30.0 mmol/L和40.0 mmol/L葡萄糖处理组Müller细胞凋亡率分别为(26.40±0.25)％、(30.19±0.16)％和(36.23±0.19)％,总体差异有统计学意义(F=294.530,P＜0.05),与正常葡萄糖组比较,30.0 mmol/L、40.0 mmol/L葡萄糖组凋亡率均明显升高,差异有统计学意义(q=0.754、0.484,P＜0.05). 结论 高浓度葡萄糖可抑制视网膜Müller细胞的生长并增加其凋亡率,葡萄糖的上述作用呈浓度依赖性.
揹景 視網膜Müller細胞具有為視網膜組織提供營養、維持視網膜的正常結構等多種生理功能,研究髮現Müller細胞的病變會導緻視網膜血管髮生相應的改變.探討高糖對視網膜Müller細胞的影響對于糖尿病視網膜病變(DR)的髮病機製研究具有重要意義. 目的 研究不同濃度的葡萄糖對體外培養的視網膜Müller細胞活性的影響. 方法 取齣生後10d清潔級SD大鼠的視網膜組織,用組織塊培養法在含質量分數20％胎牛血清的DMEM培養液中體外原代培養Müller細胞併傳代,取第3代細胞用免疫組織化學法對細胞進行鑒定.將不同濃度(5.5、30.0、40.0 mmol/L)的葡萄糖加入培養基中培養4d,MTT比色法測定各組波長570 nm處Müller細胞的吸光度(A570)值,計算各組細胞的相對存活率；採用流式細胞儀檢測各組Müller細胞的凋亡率. 結果 培養的細胞貼壁生長,呈長梭形；95％以上細胞神經膠質纖維痠性蛋白(GFAP)反應暘性.MTT比色法檢測顯示,正常葡萄糖組及30.0 mmo/L、40.0 mmol/L葡萄糖處理組Müller細胞4570值分彆為0.24±0.01、0.21±0.03和0.20±0.02,總體差異有統計學意義(F=6.755,P＜0.05).與正常葡萄糖組比較,30.0 mmol/L 、40.0 mmol/L葡萄糖處理組A570值均明顯降低,差異有統計學意義(q=0.645、0.486,P＜0.05).流式細胞儀檢測結果錶明,正常葡萄糖組、30.0 mmol/L和40.0 mmol/L葡萄糖處理組Müller細胞凋亡率分彆為(26.40±0.25)％、(30.19±0.16)％和(36.23±0.19)％,總體差異有統計學意義(F=294.530,P＜0.05),與正常葡萄糖組比較,30.0 mmol/L、40.0 mmol/L葡萄糖組凋亡率均明顯升高,差異有統計學意義(q=0.754、0.484,P＜0.05). 結論 高濃度葡萄糖可抑製視網膜Müller細胞的生長併增加其凋亡率,葡萄糖的上述作用呈濃度依賴性.
배경 시망막Müller세포구유위시망막조직제공영양、유지시망막적정상결구등다충생리공능,연구발현Müller세포적병변회도치시망막혈관발생상응적개변.탐토고당대시망막Müller세포적영향대우당뇨병시망막병변(DR)적발병궤제연구구유중요의의. 목적 연구불동농도적포도당대체외배양적시망막Müller세포활성적영향. 방법 취출생후10d청길급SD대서적시망막조직,용조직괴배양법재함질량분수20％태우혈청적DMEM배양액중체외원대배양Müller세포병전대,취제3대세포용면역조직화학법대세포진행감정.장불동농도(5.5、30.0、40.0 mmol/L)적포도당가입배양기중배양4d,MTT비색법측정각조파장570 nm처Müller세포적흡광도(A570)치,계산각조세포적상대존활솔；채용류식세포의검측각조Müller세포적조망솔. 결과 배양적세포첩벽생장,정장사형；95％이상세포신경효질섬유산성단백(GFAP)반응양성.MTT비색법검측현시,정상포도당조급30.0 mmo/L、40.0 mmol/L포도당처리조Müller세포4570치분별위0.24±0.01、0.21±0.03화0.20±0.02,총체차이유통계학의의(F=6.755,P＜0.05).여정상포도당조비교,30.0 mmol/L 、40.0 mmol/L포도당처리조A570치균명현강저,차이유통계학의의(q=0.645、0.486,P＜0.05).류식세포의검측결과표명,정상포도당조、30.0 mmol/L화40.0 mmol/L포도당처리조Müller세포조망솔분별위(26.40±0.25)％、(30.19±0.16)％화(36.23±0.19)％,총체차이유통계학의의(F=294.530,P＜0.05),여정상포도당조비교,30.0 mmol/L、40.0 mmol/L포도당조조망솔균명현승고,차이유통계학의의(q=0.754、0.484,P＜0.05). 결론 고농도포도당가억제시망막Müller세포적생장병증가기조망솔,포도당적상술작용정농도의뢰성.
Background Retinal Müller cells can offer nutrient and maintain the normal structure of retina.Researches showed that the abnormality of Müiller cells leads to retinal vascular disease.To explore the effect of high glaucoma on retinal Müller cells is of a very important significance for the study on diabetic retinopathy (DR).Objective This study was to investigate the effects of different concentrations of glucose on retinal Müller cells in vitro.Methods Retinal tissue was isolated from 1 10-day-oM clean SD rat.Mtiller cells were cultured by explant culture method and passaged in DMEM containing 20％ fetal bovine serum.The third generation of cells were obtained and identified using glial fibrillary acidic protein (GFAP) staning.Then,5.5,30.0 and 40.0 mmol/L glucose were added into the culture medium for 4 days respectively.The proliferation (A570) of Müller cells was detected by MTT,and apoptosis rate of Müller cells was calculated by flow cytometer to evaluate the effects of 5.5,30.0 and 40.0 mmol/L glucose to cell vitality.Results Cultured and passaged cells grew well with the spindle shape.The positive reactive cells were ＞95％ for GFAP.The A570 value of Müller cells was 0.24±0.01,0.21±0.03 and 0.20±0.02 in 5.5,30.0 and 40.0 mmol/L glucose group respectively,showing a significant difference among the three groups(F=6.755,P＜0.05).Compared with 5.5 mmol/L glucose group,As70 values were significantly lower in 30.0and 40.0 mmol/L glucose group (q =0.645,0.486,P ＜ 0.05).Apoptosis rates of Miiller cells were (26.40 ±0.25)％,(30.19±0.16)％ and (36.23±0.19)％ in 5.5,30.0 and 40.0mmol/L glucose groups,with a significant difference among them (F =294.530,P＜0.05),and those in 30.0 and 40.0 mmol/L glucose groups were significantly reduced in comparison with 40.0 mmol/L glucose group (q =0.754,0.484,P ＜ 0.05).Conclusions High concentration of glucose inhibits the viability and promote the apoptosis of retinal Müller cells at a concentrationdependent manner.
