中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2010年
5期
459-462
,共4页
朱冬青%许迅%郑志%邬海翔%顾青
硃鼕青%許迅%鄭誌%鄔海翔%顧青
주동청%허신%정지%오해상%고청
乳酸/激动剂%视网膜%血管内皮生长因子类%动物实验
乳痠/激動劑%視網膜%血管內皮生長因子類%動物實驗
유산/격동제%시망막%혈관내피생장인자류%동물실험
Lactic Acid/agonists%Retina%Vascular endothelial growth factors%Animal experimentation
目的 观察不同浓度乳酸对培养的大鼠视网膜血管内皮生长因子(VEGF)表达的影响.方法 2周龄Sprague-Dawley大鼠36只,根据培养液中乳酸含量分为10、20、30 mmol/L乳酸组,每组均为12只大鼠.处死大鼠,摘出眼球剥离视网膜,将视网膜置入插入式培养皿中培养24 h.培养皿中培养液分别为含10、20、30 mmol/L乳酸的Dulbecco改良Eagle培养液+2%胎牛血清.光学显微镜观察视网膜结构、实时荧光定量聚合酶链反应(RT-PCR)和蛋白质免疫印迹(Western blot)检测VEGF的表达.结果 光学显微镜组织病理学观察结果显示,视网膜层次清晰,结构完整,未见明显细胞溶解和坏死.RT-PCR检测结果显示,10、20、30 mmol/L乳酸组VEGF mRNA表达量分别为0.74±0.06、0.99±0.12、1.45±0.17;Western blot检测结果显示,10、20 mmol/L乳酸组和30 mmol/L乳酸组视网膜VEGF表达量分别为0.34±0.15、0.54±0.16、0.93±0.23.RT-PCT和Western blot检测结果均显示30 mmol/L乳酸组较10 mmol/L乳酸组VEGF表达明显升高.结论 乳酸诱导视网膜VEGF表达有浓度依赖性.
目的 觀察不同濃度乳痠對培養的大鼠視網膜血管內皮生長因子(VEGF)錶達的影響.方法 2週齡Sprague-Dawley大鼠36隻,根據培養液中乳痠含量分為10、20、30 mmol/L乳痠組,每組均為12隻大鼠.處死大鼠,摘齣眼毬剝離視網膜,將視網膜置入插入式培養皿中培養24 h.培養皿中培養液分彆為含10、20、30 mmol/L乳痠的Dulbecco改良Eagle培養液+2%胎牛血清.光學顯微鏡觀察視網膜結構、實時熒光定量聚閤酶鏈反應(RT-PCR)和蛋白質免疫印跡(Western blot)檢測VEGF的錶達.結果 光學顯微鏡組織病理學觀察結果顯示,視網膜層次清晰,結構完整,未見明顯細胞溶解和壞死.RT-PCR檢測結果顯示,10、20、30 mmol/L乳痠組VEGF mRNA錶達量分彆為0.74±0.06、0.99±0.12、1.45±0.17;Western blot檢測結果顯示,10、20 mmol/L乳痠組和30 mmol/L乳痠組視網膜VEGF錶達量分彆為0.34±0.15、0.54±0.16、0.93±0.23.RT-PCT和Western blot檢測結果均顯示30 mmol/L乳痠組較10 mmol/L乳痠組VEGF錶達明顯升高.結論 乳痠誘導視網膜VEGF錶達有濃度依賴性.
목적 관찰불동농도유산대배양적대서시망막혈관내피생장인자(VEGF)표체적영향.방법 2주령Sprague-Dawley대서36지,근거배양액중유산함량분위10、20、30 mmol/L유산조,매조균위12지대서.처사대서,적출안구박리시망막,장시망막치입삽입식배양명중배양24 h.배양명중배양액분별위함10、20、30 mmol/L유산적Dulbecco개량Eagle배양액+2%태우혈청.광학현미경관찰시망막결구、실시형광정량취합매련반응(RT-PCR)화단백질면역인적(Western blot)검측VEGF적표체.결과 광학현미경조직병이학관찰결과현시,시망막층차청석,결구완정,미견명현세포용해화배사.RT-PCR검측결과현시,10、20、30 mmol/L유산조VEGF mRNA표체량분별위0.74±0.06、0.99±0.12、1.45±0.17;Western blot검측결과현시,10、20 mmol/L유산조화30 mmol/L유산조시망막VEGF표체량분별위0.34±0.15、0.54±0.16、0.93±0.23.RT-PCT화Western blot검측결과균현시30 mmol/L유산조교10 mmol/L유산조VEGF표체명현승고.결론 유산유도시망막VEGF표체유농도의뢰성.
Objective To investigate if lactic acid can promote the expression of vascular endothelial growth factor (VEGF) in the rat retinal explants. Methods The retinas of two-week neonatal SD rats were placed onto the culture plate inserts and incubated with Dulbecco's modified Eagle's medium (DMEM) plus 2% fetal bovine serum (FBS) containing 10, 20, 30 mmol/L of lactic acid, respectively. Each group had 24 retinas. At 24 hours after incubation, the retinas were sectioned for light microscopy and the expression of VEGF was measured by real time PCR and Western blot. Results The cultured retinas maintained intact construction, and no cytolysis and apoptosis were observed under light microscope. RT-PCR showed the levels of VEGF mRNA were 0.74 ± 0.06 for 10 mmol/L lactic acid group, 0. 99 ± 0. 12 for 20 mmol/L group, and 1.45±0. 17 for 30 mmol/L group respectively. VEGF expression was 0. 34±0. 15 for 10 mmol/L,0. 54±0. 16 for 20 mmol/L, and 0. 93±0. 23 for 30 mmol/L group respectively by Western blot. Both PCR and Western blot showed 30 mmol/L of lactic acid significantly increased the levels of VEGF mRNA and VEGF expression. Conclusion The induction of retinal VEGF by lactic acid is concentration-dependent.
