磷酸果糖激酶-2/果糖-2,6-二磷酸酶3在增生型糖尿病性视网膜病变患者玻璃体和血清中的表达及其相关性分析
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摘要
目的定量检测增生型糖尿病性视网膜病变(proliferative diabetic retinopathy,PDR)患者玻璃体和血清中磷酸果糖激酶-2/果糖-2,6-二磷酸酶3(phospofructokinase-2/fructose-2,6-bisphosphatase 3,PFKFB3)和血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)的表达水平,探讨PFKFB3在PDR患者中可能的作用机制。方法纳入确诊为PDR拟行单眼玻璃体切割术的患者42例(42眼)作为试验组,并根据术前是否行玻璃体内注射抗VEGF药物进一步将试验组分为非注药组和注药组,纳入同期诊断为全层黄斑裂孔(full thickness macular hole,FTMH)及黄斑前膜(preretinal membrane of the macula,PRMM)的20例(20眼)患者作为对照组。收集各组患者的玻璃体和血清标本,定量检测各组标本中的PFKFB3和VEGF表达水平,并进行统计学分析。结果试验组患者玻璃体中PFKFB3水平为(463. 17±381. 28) ng·L~(-1),明显高于对照组[(158. 43±86. 88)ng·L~(-1)](t=4. 919,P <0. 001),试验组血清中PFKFB3水平为(153. 45±83. 78) ng·L~(-1),明显高于对照组[(92. 72±32. 42)ng·L~(-1)](t=4. 098,P <0. 001)。非注药组患者玻璃体中PFKFB3水平为(797. 29±387. 44) ng·L~(-1),明显高于注药组[(257. 56±181. 49) ng·L~(-1)](t=5. 230,P <0. 001),而非注药组血清中PFKFB3水平与注药组差异无统计学意义(t=0. 679,P=0. 501)。非注药、注药组和对照组患者玻璃体与血清中的PFKFB3水平无相关性(非注药组:r=0. 194,P=0. 471;注药组:r=0. 071,P=0. 731;对照组:r=0. 254,P=0. 279)。试验组患者玻璃体中VEGF水平为(1713. 50±1386. 90) ng·L~(-1),明显高于对照组[(205. 52±92. 93) ng·L~(-1)](t=7. 014,P <0. 001);试验组血清中VEGF水平为(224. 98±168. 08) ng·L~(-1),明显高于对照组[(86. 80±36. 51) ng·L~(-1)](t=5. 082,P <0. 001)。非注药组患者玻璃体中VEGF水平为(3399. 72±510. 06) ng·L~(-1),明显高于注药组[(675. 82±242. 57) ng·L~(-1)](t=20. 014,P <0. 001),非注药组血清中VEGF水平为(373. 40±174. 23) ng·L~(-1),明显高于注药组[(133. 65±73. 10) ng·L~(-1)](t=5. 228,P <0. 001)。非注药、注药组和对照组患者玻璃体与血清中的VEGF水平均存在正相关关系(非注药组:r=0. 952,P <0. 001;注药组:r=0. 423,P=0. 031;对照组:r=0. 776,P <0. 001)。非注药组、注药组和对照组患者玻璃体中PFKFB3与VEGF水平均存在正相关关系(非注药组:r=0. 865,P <0. 001;注药组:r=0. 587,P=0. 002;对照组:r=0. 807,P <0. 001),而在血清中仅注药组的PFKFB3与VEGF水平存在正相关关系(r=0. 444,P=0. 023)。结论PFKFB3可能参与了PDR的发生发展过程; VEGF可能通过上调PFKFB3的表达而参与PDR的发生发展。
Objective To discuss the possible acting mechanism of phospofructokinase-2/fructose-2,6-bisphosphatase 3(PFKFB3) in the occurrence and development of proliferative diabetic retinopathy(PDR) by means of quantitatively measuring the levels of PFKFB3 and vascular endothelial growth factor(VEGF) in vitreous and serum of patients with PDR.Methods Totally 42 patients(42 eyes) who were diagnosed with PDR and planned to accept vitrectomy in a single eye were collected as the experimental group.The experimental group was further divided into non-drug-injection and drug-injection group according to whether anti-VEGF drugs were injected into vitreous cavity before operation.Totally 20 patients(20 eyes) who were diagnosed with full thickness macular hole(FTMH) and preretinal membrane of the macula(PRMM) were selected as the control group.Vitreous and serum specimens of patients in each group were collected,then PFKFB3 and VEGF expression levels were detected in vitreous and serum specimens.Statistical software was used to perform statistical analysis for data in each group.Results The levels of PFKFB3 in vitreous of the experimental group was(463.17±381.28)ng·L~(-1),significantly higher than that of the control group[(158.43±86.88)ng·L~(-1)](t=4.919,P<0.001).The levels of PFKFB3 in serum of the experimental group was(153.45±83.78)ng·L~(-1),higher than that of the control group[(92.72±32.42)ng·L~(-1)](t=4.098,P<0.001).The level of PFKFB3 in vitreous of non-drug-injection group was(797.29±387.44)ng·L~(-1),significantly higher than that of the drug-injection group[(257.56±181.49)ng·L~(-1)](t=5.230,P<0.001),but there was no difference between non-drug-injection and drug-injection group in serum(t=0.679,P=0.501).The levels of PFKFB3 has no correlation in vitreous and serum among the non-drug-injection group,drug-injection group and control group(non-drug-injection group:r=0.194,P=0.471;drug-injection group r=0.071,P=0.731;control group r=0.254,P=0.279).The levels of VEGF in vitreous in experimental group was(1713.50±1386.90)ng·L~(-1),higher than that of the control group[(205.52±92.93)ng·L~(-1)](t=7.014,P<0.001).The levels of VEGF in serum in experimental group was(224.98±168.08)ng·L~(-1),significantly higher than that of the control group [(86.80±36.51)ng·L~(-1)](t=5.082,P<0.001).The levels of VEGF in vitreous in non-drug-injection group was(3399.72±510.06)ng·L~(-1),evidently higher than that of the drug-injection group [(675.82±242.57)ng·L~(-1)](t=20.014,P<0.001).The levels of VEGF in serum in non-drug-injection group was(373.40±174.23)ng·L~(-1),higher than that of the drug-injection group[(133.65±73.10)ng·L~(-1)](t=5.228,P<0.001).The levels of VEGF has a positive correlation in vitreous and serum among the non-drug-injection group,drug-injection group and control group(non-drug-injection group:r=0.952,P<0.001;drug-injection group:r=0.423,P=0.031;control group:r=0.776,P<0.001).The levels of PFKFB3 and VEGF in vitreous body has a positive correlation among non-drug-injection,drug-injection group and control group(non-drug-injection group:r=0.865,P<0.001;drug-injection group:r=0.587,P=0.002;control group:r=0.807,P<0.001),while there was a positive correlation between PFKFB3 and VEGF only in serum of drug-injection group(r=0.444,P=0.023).Conclusion PFKFB3 may be involved in the occurrence and development of PDR,and VEGF may be involved in PDR by up-regulating the expression of PFKFB3.
Objective To discuss the possible acting mechanism of phospofructokinase-2/fructose-2,6-bisphosphatase 3(PFKFB3) in the occurrence and development of proliferative diabetic retinopathy(PDR) by means of quantitatively measuring the levels of PFKFB3 and vascular endothelial growth factor(VEGF) in vitreous and serum of patients with PDR.Methods Totally 42 patients(42 eyes) who were diagnosed with PDR and planned to accept vitrectomy in a single eye were collected as the experimental group.The experimental group was further divided into non-drug-injection and drug-injection group according to whether anti-VEGF drugs were injected into vitreous cavity before operation.Totally 20 patients(20 eyes) who were diagnosed with full thickness macular hole(FTMH) and preretinal membrane of the macula(PRMM) were selected as the control group.Vitreous and serum specimens of patients in each group were collected,then PFKFB3 and VEGF expression levels were detected in vitreous and serum specimens.Statistical software was used to perform statistical analysis for data in each group.Results The levels of PFKFB3 in vitreous of the experimental group was(463.17±381.28)ng·L~(-1),significantly higher than that of the control group[(158.43±86.88)ng·L~(-1)](t=4.919,P<0.001).The levels of PFKFB3 in serum of the experimental group was(153.45±83.78)ng·L~(-1),higher than that of the control group[(92.72±32.42)ng·L~(-1)](t=4.098,P<0.001).The level of PFKFB3 in vitreous of non-drug-injection group was(797.29±387.44)ng·L~(-1),significantly higher than that of the drug-injection group[(257.56±181.49)ng·L~(-1)](t=5.230,P<0.001),but there was no difference between non-drug-injection and drug-injection group in serum(t=0.679,P=0.501).The levels of PFKFB3 has no correlation in vitreous and serum among the non-drug-injection group,drug-injection group and control group(non-drug-injection group:r=0.194,P=0.471;drug-injection group r=0.071,P=0.731;control group r=0.254,P=0.279).The levels of VEGF in vitreous in experimental group was(1713.50±1386.90)ng·L~(-1),higher than that of the control group[(205.52±92.93)ng·L~(-1)](t=7.014,P<0.001).The levels of VEGF in serum in experimental group was(224.98±168.08)ng·L~(-1),significantly higher than that of the control group [(86.80±36.51)ng·L~(-1)](t=5.082,P<0.001).The levels of VEGF in vitreous in non-drug-injection group was(3399.72±510.06)ng·L~(-1),evidently higher than that of the drug-injection group [(675.82±242.57)ng·L~(-1)](t=20.014,P<0.001).The levels of VEGF in serum in non-drug-injection group was(373.40±174.23)ng·L~(-1),higher than that of the drug-injection group[(133.65±73.10)ng·L~(-1)](t=5.228,P<0.001).The levels of VEGF has a positive correlation in vitreous and serum among the non-drug-injection group,drug-injection group and control group(non-drug-injection group:r=0.952,P<0.001;drug-injection group:r=0.423,P=0.031;control group:r=0.776,P<0.001).The levels of PFKFB3 and VEGF in vitreous body has a positive correlation among non-drug-injection,drug-injection group and control group(non-drug-injection group:r=0.865,P<0.001;drug-injection group:r=0.587,P=0.002;control group:r=0.807,P<0.001),while there was a positive correlation between PFKFB3 and VEGF only in serum of drug-injection group(r=0.444,P=0.023).Conclusion PFKFB3 may be involved in the occurrence and development of PDR,and VEGF may be involved in PDR by up-regulating the expression of PFKFB3.
引文
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[5] DE BOCK K D,GEORGIADOU M,SCHOORS S,KUCHNIO A,WONG B W,CANTELMO A R,et al.Role of PFKFB3-driven glycolysis in vessel sprouting[J].Cell,2013,154(3):651-663.
[6] KRUTZFELDT A,SPAHR R,MERTENS S,SIEGMUND B,PIPER H M.Metabolism of exogenous substrates by coronary endothelial cells in culture[J].J Mol Cell Cardio,1990,22(12):1393-1404.
[7] XU Y,AN X,GUO X,HABTETSION T G,WANG Y,XU X,et al.Endothelial PFKFB3 plays a critical role in angiogenesis[J].Arterioscler Thromb Vasc Biol,2014,34(6):1231-1239.
[8] WANG H,ZHANG H,SUN P,GU F,FU B,et al.Determination of vascular endothelial growth factor and tumor necrosis factor-alpha in vitreous fluid of patients with retinal vascular disease[J].Chin J Ocul Fundus Dis,2012,28(5):519-521.王欢,张含,孙鹏,谷峰,董洁,傅博,等.视网膜血管性疾病患者玻璃体液中血管内皮生长因子和肿瘤坏死因子-α的含量测定[J].中华眼底病杂志,2012,28(5):519-521.
[9] SYDOROVA M,LEE M S.Vascular endothelial growth factor levels in vitreous and serum of patients with either proliferative diabetic retinopathy or proliferative vitreoretinopathy[J].Ophthalmic Res,2005,37(4):188-190.
[10] BAHARIVAND N,ZARGHAMI N,PANAHI F,GHAFARI M Y D,FARD A M,MOHAJERI A.Relationship between vitreous and serum vascular endothelial growth factor levels,control of diabetes and microalbuminuria in proliferative diabetic retinopathy[J].Clin Ophthalmol,2012,6:185-191.
[11] MA Y,ZHANG Y,ZHAO T,JIANG Y R.Vascular endothelial growth factor in plasma and vitreous fluid of patients with proliferativediabetic retinopathy patients after intravitreal injection of bevacizumab[J].Am J Ophthalmol,2012,153(2):307-313.
[12] SAN J J,YUAN Z L,CAO G P.Changes of serum vascular endothelial growth factor in diabetic retinopathy patients after intravitreal injection of bevacizumab Avastin[J].J Clin Ophthalmol,2014,22(4):333-335.单俊杰,袁志兰,曹国平.玻璃体腔注射贝伐单抗Avastin后糖尿病视网膜患者血清中VEGF含量的变化研究[J].临床眼科杂志,2014,22(4):333-335.
[13] POTENTE M,GERHARDT H,CARMELIET P.Basic and therapeutic aspects of angiogenesis[J].Cell,2011,146(6):873-887.
[14] EILKEN H M,ADAMS R H.Dynamics of endothelial cell behavior in sprouting angiogenesis[J].Curr Opin Cell Biol,2010,22(5):617-625.
[15] ALMEIDA A,BOLANOS J P,MONCADA S.E3 ubiquitin ligase APC/C-Cdh1 accounts for the Warburg effect by linking glycolysis to cell proliferation[J].Proc Natl Acad Sci U S A,2010,107(2):738-741.
[2] SCHOORS S,DE BOCK K K,CANTELMO A R,GEORGIADOU M,GHESQU-IERE B,CAUWENBERGHS S,et al.Partial and transient reduction of glycolysis by PFKFB3 blockade reduces pathological angiogenesis[J].Cell Metabolism,2014,19(1):37-48.
[3] MINCHENKO A,LESHCHINSKY I,OPENTANOVA I,SANG N,SRINIVAS V,ARMSTEAD V,et al.Hypoxia-inducible factor-1-mediated expression of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-3 (PFKFB3) gene.Its possible role in the Warburg effect[J].J Biol Chem,2002,277(8):6183-6187.
[4] MARSIN A S,BOUZIN C,BERTRAND L,HUE L.The stimulation of glycolysis by hypoxia in activated monocytes is mediated by AMP-activated protein kinase and inducible 6-phosphofructo-2-kinase[J].J Biol Chem,2002,277(34):30778-30783.
[5] DE BOCK K D,GEORGIADOU M,SCHOORS S,KUCHNIO A,WONG B W,CANTELMO A R,et al.Role of PFKFB3-driven glycolysis in vessel sprouting[J].Cell,2013,154(3):651-663.
[6] KRUTZFELDT A,SPAHR R,MERTENS S,SIEGMUND B,PIPER H M.Metabolism of exogenous substrates by coronary endothelial cells in culture[J].J Mol Cell Cardio,1990,22(12):1393-1404.
[7] XU Y,AN X,GUO X,HABTETSION T G,WANG Y,XU X,et al.Endothelial PFKFB3 plays a critical role in angiogenesis[J].Arterioscler Thromb Vasc Biol,2014,34(6):1231-1239.
[8] WANG H,ZHANG H,SUN P,GU F,FU B,et al.Determination of vascular endothelial growth factor and tumor necrosis factor-alpha in vitreous fluid of patients with retinal vascular disease[J].Chin J Ocul Fundus Dis,2012,28(5):519-521.王欢,张含,孙鹏,谷峰,董洁,傅博,等.视网膜血管性疾病患者玻璃体液中血管内皮生长因子和肿瘤坏死因子-α的含量测定[J].中华眼底病杂志,2012,28(5):519-521.
[9] SYDOROVA M,LEE M S.Vascular endothelial growth factor levels in vitreous and serum of patients with either proliferative diabetic retinopathy or proliferative vitreoretinopathy[J].Ophthalmic Res,2005,37(4):188-190.
[10] BAHARIVAND N,ZARGHAMI N,PANAHI F,GHAFARI M Y D,FARD A M,MOHAJERI A.Relationship between vitreous and serum vascular endothelial growth factor levels,control of diabetes and microalbuminuria in proliferative diabetic retinopathy[J].Clin Ophthalmol,2012,6:185-191.
[11] MA Y,ZHANG Y,ZHAO T,JIANG Y R.Vascular endothelial growth factor in plasma and vitreous fluid of patients with proliferativediabetic retinopathy patients after intravitreal injection of bevacizumab[J].Am J Ophthalmol,2012,153(2):307-313.
[12] SAN J J,YUAN Z L,CAO G P.Changes of serum vascular endothelial growth factor in diabetic retinopathy patients after intravitreal injection of bevacizumab Avastin[J].J Clin Ophthalmol,2014,22(4):333-335.单俊杰,袁志兰,曹国平.玻璃体腔注射贝伐单抗Avastin后糖尿病视网膜患者血清中VEGF含量的变化研究[J].临床眼科杂志,2014,22(4):333-335.
[13] POTENTE M,GERHARDT H,CARMELIET P.Basic and therapeutic aspects of angiogenesis[J].Cell,2011,146(6):873-887.
[14] EILKEN H M,ADAMS R H.Dynamics of endothelial cell behavior in sprouting angiogenesis[J].Curr Opin Cell Biol,2010,22(5):617-625.
[15] ALMEIDA A,BOLANOS J P,MONCADA S.E3 ubiquitin ligase APC/C-Cdh1 accounts for the Warburg effect by linking glycolysis to cell proliferation[J].Proc Natl Acad Sci U S A,2010,107(2):738-741.