中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2008年
7期
640-644
,共5页
邬海翔%夏欣%刘堃%ZHENG Zhi%朱冬青%XU Xun%顾青
鄔海翔%夏訢%劉堃%ZHENG Zhi%硃鼕青%XU Xun%顧青
오해상%하흔%류곤%ZHENG Zhi%주동청%XU Xun%고청
胰岛素%视网膜血管%内皮细胞%血管内皮生长因子A%糖尿病视网膜病变
胰島素%視網膜血管%內皮細胞%血管內皮生長因子A%糖尿病視網膜病變
이도소%시망막혈관%내피세포%혈관내피생장인자A%당뇨병시망막병변
Insulin%Retinal vessels%Endothelial cells%Vascular endothelial growth factor A%Diabetic retinopathy
目的 探讨胰岛素、糖浓度及其两者的联合作用对牛视网膜微血管内皮(BRE)细胞血管内皮生长因子(VEGF)表达的影响.方法 对照实验研究.采用选择性培养方法,培养BRE细胞,传代后分别在正常糖(5 mmol/L)或高糖(30 mmol/L)浓度下培养3 d,甘露醇平衡渗透压,血清饥饿12 h,给予或不给予100 nmol/L胰岛素作用24 h.实时荧光定量检测VEGF mRNA表达水平;人脐静脉血管内皮细胞增殖法、免疫荧光检测法、免疫印迹法检测VEGF蛋白表达水平.采用SPSS 12.0统计学软件对数据进行分析,胰岛素、糖浓度及其交互作用对BRE细胞的VEGF mRNA和VEGF蛋白表达水平的影响,采用2×2析因设计定量资料方差分析,以P<0.05作为差异有统计学意义.结果 胰岛素或高糖浓度可以显著提高BRE细胞的VEGFmRNA(F=5.67,9.04;均P<0.05)和VEGF蛋白(F=5.50,5.57;均P<0.05)表达水平,但胰岛素联合高糖浓度的作用较其单独作用减弱.结论 高糖浓度下可以降低胰岛素诱导的VEGF表达水平,因此,VEGF可能不是胰岛素治疗导致的糖尿病视网膜病变短期恶化的主要因素.
目的 探討胰島素、糖濃度及其兩者的聯閤作用對牛視網膜微血管內皮(BRE)細胞血管內皮生長因子(VEGF)錶達的影響.方法 對照實驗研究.採用選擇性培養方法,培養BRE細胞,傳代後分彆在正常糖(5 mmol/L)或高糖(30 mmol/L)濃度下培養3 d,甘露醇平衡滲透壓,血清饑餓12 h,給予或不給予100 nmol/L胰島素作用24 h.實時熒光定量檢測VEGF mRNA錶達水平;人臍靜脈血管內皮細胞增殖法、免疫熒光檢測法、免疫印跡法檢測VEGF蛋白錶達水平.採用SPSS 12.0統計學軟件對數據進行分析,胰島素、糖濃度及其交互作用對BRE細胞的VEGF mRNA和VEGF蛋白錶達水平的影響,採用2×2析因設計定量資料方差分析,以P<0.05作為差異有統計學意義.結果 胰島素或高糖濃度可以顯著提高BRE細胞的VEGFmRNA(F=5.67,9.04;均P<0.05)和VEGF蛋白(F=5.50,5.57;均P<0.05)錶達水平,但胰島素聯閤高糖濃度的作用較其單獨作用減弱.結論 高糖濃度下可以降低胰島素誘導的VEGF錶達水平,因此,VEGF可能不是胰島素治療導緻的糖尿病視網膜病變短期噁化的主要因素.
목적 탐토이도소、당농도급기량자적연합작용대우시망막미혈관내피(BRE)세포혈관내피생장인자(VEGF)표체적영향.방법 대조실험연구.채용선택성배양방법,배양BRE세포,전대후분별재정상당(5 mmol/L)혹고당(30 mmol/L)농도하배양3 d,감로순평형삼투압,혈청기아12 h,급여혹불급여100 nmol/L이도소작용24 h.실시형광정량검측VEGF mRNA표체수평;인제정맥혈관내피세포증식법、면역형광검측법、면역인적법검측VEGF단백표체수평.채용SPSS 12.0통계학연건대수거진행분석,이도소、당농도급기교호작용대BRE세포적VEGF mRNA화VEGF단백표체수평적영향,채용2×2석인설계정량자료방차분석,이P<0.05작위차이유통계학의의.결과 이도소혹고당농도가이현저제고BRE세포적VEGFmRNA(F=5.67,9.04;균P<0.05)화VEGF단백(F=5.50,5.57;균P<0.05)표체수평,단이도소연합고당농도적작용교기단독작용감약.결론 고당농도하가이강저이도소유도적VEGF표체수평,인차,VEGF가능불시이도소치료도치적당뇨병시망막병변단기악화적주요인소.
Objective To investigate the effect of insulin or (and) high glucose on the vascular endothelial growth factor (VEGF) expression in bovine retinal microvascular endothelial cells (BREC).Methods It was a experimental study. BREC were cultured with selective culture media. Passaged cells were cultured in normal (5 mmol/L) or high glucose (30 mmol/L) medium for 3 days. Equimolar mannitol was supplemented for osmotic controls. Cells were serum-deprived overnight in Dulbeceo's modified Eagle's medium containing 0.2% w/v bovine serum albumin and then incubated in the absence or presence of 100nmol/L insuhn for 24 hrs. Cells were harvested and the expression of VEGF mRNA was analyzed by Realtime PCR. VEGF protein level was determined by HUVEC proliferation assay, immunofluorescence and western-blot. Results Insulin or (and) high glucose significantly increased VEGF mRNA ( F = 5.67,9.04 ;P <0.05) and protein (F=5.50,5.57;P<0.05) expression. However, the combined effect of insulin and high glucose is weak in contrast to insulin or high glucose alone. Conclusions High glucose attenuated insulin-induced VEGF expression. Therefore, VEGF may be not the major factor that lead to transient worsening of diabetic retinopathy after acute insulin therapy in diabetes.
