摘要
目的:糖尿病(DM)是以糖代谢紊乱为主的常见全身性疾病,患病率逐年提高。DM可引起眼部多种并发症,近年来的临床资料观察到DM患者出现眼干涩、异物感、烧灼感的症状较普遍。本实验制作链脲佐菌素(STZ)大鼠DM动物模型,通过测量泪膜破裂时间(BUT)和基础泪液分泌量(Schirmer-Ⅰ试验)观察DM对照组(DM组)及氨基胍(AG)治疗组(治疗组)大鼠眼表稳定性的改变;并通过观察泪腺、结膜和角膜的组织病理学改变及肿瘤坏死因子-α(TNF-α)、核转录因子-KBp65(NF-KBp65)、糖基化终产物受体(RAGE)的免疫组织化学染色的改变,探讨DM造成的眼表组织改变、发病机制以及AG对DM眼表病变的治疗效果,从而为DM眼表改变提供病理学基础,并为其治疗DM眼表疾病提供新的线索和理论依据。
方法:
1实验分组
将30只180-200g雄性Wistar大鼠随机分为3组:正常对照组(正常组)10只,DM对照组(DM组)10只,AG治疗组(治疗组)10只。
2 DM动物模型的制备
禁食12h后,对三组大鼠碘伏消毒腹腔注射部位,DM组和治疗组大鼠腹腔一次性注射10g/L的STZ 65mg/kg(溶于20mmol/L枸橼酸钠缓冲液,pH4.5),之后72h和实验中每4wk检测一次血糖,血糖始终大于16.7mmol/L者纳入实验。正常组腹腔一次性注射等量的20mmol/L枸橼酸钠缓冲液(pH4.5)。
3给药方式
从腹腔注射STZ 72h开始,对三组大鼠碘伏消毒腹腔注射部位,治疗组给予腹腔注射新鲜配制的0.2%AG(80mg/kg.d,用25mmol/L、pH为7.4的磷酸钠缓冲液配制),DM组和正常组分别给予腹腔注射同等剂量的磷酸钠缓冲液(pH为7.4),1/d。
4观察指标
每日观察各组大鼠的饮水量、饮食量和尿量。每周使用天平称大鼠体重并记录。注射STZ 72h后和每4周大鼠尾端75%酒精消毒后针刺取血,检测血糖。12wk行BUT和Schirmer-Ⅰ试验,脱颈椎法分别处死大鼠每组各10只,立即摘除泪腺、结膜和角膜,用10g/L中性多聚甲醛固定备HE染色和免疫组织化学染色用。
结果:
1正常组大鼠体重随时间延长而逐渐增加;DM组大鼠和治疗组大鼠在STZ注射后2-12wk体重明显低于正常组,差异有统计学意义(p<0.05),在各时间点,DM组和治疗组大鼠体重差异无统计学意义(p>0.05)(Fig 1)。
2 DM组和治疗组大鼠血糖在注射STZ 72h后明显高于正常组,差异具有统计学意义(P<0.01);治疗组和DM组在各时间点大鼠血糖差异无统计学意义(P>0.05)(Fig 2)。
3 BUT和Schirmer-Ⅰ试验值的测定:与正常组比较,DM组降低,有统计学意义(p<0.01),治疗组亦降低,有统计学意义(p<0.05);治疗组较DM组明显升高,有统计学意义(p<0.01)(Table 1)。
4泪腺、结膜和角膜组织病理学变化:DM组大鼠泪腺腺泡变小,细胞核固缩、深染,细胞分界不清,结膜及角膜损害,表现为结膜杯状细胞减少,角膜上皮和基质层水肿;治疗组大鼠泪腺、结膜和角膜病变程度较DM组大鼠减轻,但较正常组大鼠有明显的差异(Fig 3-11)。
5 TNF-α、NF-KBp65和RAGE在各组大鼠泪腺、结膜和角膜的变化:TNF-α、NF-KBp65和RAGE在各组大鼠泪腺、结膜和角膜的胞浆中均有表达;与正常组比较,DM组和治疗组TNF-α、NF-KBp65和RAGE明显升高,差异有统计学意义(p<0.01);治疗组与DM组比较,TNF-α、NF-KBp65和RAGE均降低,差异有统计学意义(p<0.01)(Fig 12-38,Table 2-4)。且在DM组TNF-α、NF-KBp65和RAGE三者之间互相呈正相关关系(Table 5)。
结论:
1糖尿病与干眼有关。
2糖尿病眼表病变与蛋白质糖基化反应有关。
3氨基胍对糖尿病引起的眼表病变有一定的治疗作用,其作用机制与影响蛋白质糖基化反应,降低TNF-α、NF-KBp65和RAGE的含量有关;且氨基胍对血糖和体重无影响。
Objective:Diabetic mellitus is a common general disease mainly manifestating by glycometabolism disorders whose prevalence is rising year by year. It can cause many complications in eyes. The recent clinical data indicates that the symptom of Dryness of eye, foreign body sensation and burning sensation appeare widespread in DM patients. The experimental model of diabetic rats are induced by intravenous administration of streptozotocin, then we observe the change of ocular surface by Schirmer-Ⅰtest and tear film break up time(BUT) measurement; The histopathological changes and the expressions of tumor necrosis factor alpha(TNF-α), nuclear factor-KBp65(NF-KBp65), receptor for advanced glycation end products(RAGE)of the cornea, conjunctiva and lacrimal gland are observed in the diabetic rats, then we conform diabetic mellitus, the effect of aminoguanidine on it, in order to provide pathological and theoretical basis for treating diabetic ocular surface diseases.
Methods:
1 Method of grouping
30 male Wistar rats that weighted 180-200g,were randomly divided into 3 groups: normal control group (normal group) , diabetic control group (diabetic group) and aminoguanidine treated group (treated group).
2 Preparation of diabetic animal model
After fasting 12h, three groups’rats were sterilized the site of intraperitoneal injection with Iodophors.Then rats of diabetic and treated group received a single 65mg/kg intraperitonea injection of 10g/L streptozotocin(STZ) dissolved in 20mmol/L sodium citrate buffer (pH 4.5). Detect blood glucose by 72h and per 4 weeks of experiment after STZ injection, then animals that blood levels more than 16.7mmol/L all the time were selected for study. In addition, 10 normal group rats received 20mmol/L sodium citrate buffer (pH 4.5).
3 Ways of administration
72h after STZ injection, three groups’rats were sterilized the site of intraperitoneal injection with Iodophors. Then treated group received injection of a freshly 80mg/kg.d solution of 0.2% aminoguanidine (25mmol/L phosphate buffered solution, pH7.4) once a day. In addition, diabetic and normal groups received 25mmol/L phosphate buffered solution (pH7.4) alone.
4 Indexes to be observed
The volume of drinking, appetite and the urinary volume were observed every day. Body weights of rats were measured weekly. Every 4 weeks during the experiment and 72h after STZ injection, blood glucose of rats were detected. All rats took the Schirmer-Ⅰtest and BUT on the 12th week. Then 10 rats of each group were sacrificed, cornea, conjunctiva and lacrimal gland were obtained and fixed by 10g/L formalin for histopathology and immunohistochemistry.
Results:
1 Body weight of normal group rats increased gradually. 2-12wk after injection of STZ, the weight was significantly decreased in diabetic and treated groups compared with normal group (p<0.05). Through all facets of the study, the difference of weight beween diabetic and treated groups rats were non-significant(P>0.05) (Fig 1).
2 Blood glucose was significantly higher in diabetic and treated groups 72h after injection of STZ,compared with control group (p<0.01); Through all facets of the study, the difference of blood glucose between diabetic and treated groups rats were non-significant(P>0.05)(Fig 2).
3 The test values of BUT and Schirmer-Ⅰ: compared with normal group,they were significantly lower in diabetic group(p<0.01),still significantly lower in treated group(p< 0.05); compared with diabetic group, they were significantly greater in treated group(p<0.01) (Table 1).
4 Histopathological changes of lacrimal gland, conjunctiva and cornea: Lacrimal glandular follieles of diabetic group shrinked, caryon were pyknosised and anachromasised. Fibers were prolifered, tissues were confused.Goblet cell lost and the basal epithelial cells and the stroma of diabetic groups’cornea were swollen. All of the changes were significantly lightener in treated group than in diabetic group, but still differed from normal group (Fig 3-11).
5 Expressions of TNF-α, NF-KBp65 and RAGE in lacrimal gland、conjunctiva and cornea: TNF-α, NF-KBp65 and RAGE are all expressed in the kytoplasm of lacrimal gland、conjunctiva and cornea. Compared with normal group, expressions of TNF-α,NF-KBp65 and RAGE were significantly greater in diabetic and treated groups (p<0.01). But compared with diabetic group, they were significantly lower(p<0.01)(Fig 12-20, Table 2-4). In addition, there were positive correlations in TNF-αand NF-KBp65, TNF-αand RAGE, NF-KBp65 and RAGE (Table 5).
Conclusions:
1 Diabetic mellitus is related with dry eye.
2 Diabetic mellitus ocular disease is concerned with protein glycosylation.
3 The effect of AG is significant in treating diabetic mellitus ocular diseases, and its mechanism of action is concerned with protein glycosylation and the decrease of TNF-α, NF-KBp65 and RAGE. In addition, it has no influence on body weight and blood glucose.
引文
1 Nelson JD,Wright JC. Conjunctival goblet cell densities in ocular surface disease. Arch Ophthalmol,1984,102(7): 1049-1051
2 李凤鸣.中华眼科学[M].北京:人民卫生出版社.2005,1153
3 吴晓梅,黄雨梅,姚先莹等.干燥综合征患者泪腺组织的形态及免疫组化研究[J].华西医大学报,1997,28(3): 284-287
4 Seifart, Strempel. Trockenes auge and Diabetes mellitus. Ophthalmology.1994, 91(2):235-239
5 Aragona P. Tear film stability and osmolarity in diabetes mellitus. Invest Ophthalmol Vis Sci,1999,40(SuPPI):542
6 Inoue K,Kato S,Ohara C,Numaga J,Amano S,Oshika T. Ocular and Systemic factors relevant to diabetic keratoepitheliopathy. Cornea,2001,20(8)798-801
7 Rumble JR,Cooper ME,Soulis T. Vascular hypertrophy in experimental diabetes. Role of advanced glycaton end products. J Clin Invest,1997,99(5): 1106-1127
8 Arendt CW, Albrecht B, Soos TJ, et al. Protein kinase CQ signaling from the centerof Ⅱle T-cell synapse[J]. Curt Opin Immunol, 2002, 14(3):323-330
9 Sakurai S,Yonekura H,YamamotoY,Watanabe T,Tanaka N,Li H,et al. The AGE-RAGE system and diabetic nephropathy. J Am Soc Nephrol,2003, 14 (8 SuPP13):5259-5263
10 Beppu H, Nakamura M, Katakami C, et al. Expression of Mn-SOD mRNA in streptozotocin-induced diabetic rat cornea by in situ hybridization[J]. Nippon Ganka Gakkai Zasshi. 1996,100(8): 599-604
11 Yucel I, Yucel G, Akar Y, et al. Transmission electron microscopy and autofluorescence findings in the cornea of diabetic rats treated with Aminoguanidine[J]. Can J Ophthalmol. 2006,41(1):60-66
12 Forbes JM, Thallas V, Thomas MC, et al. The breakdown of preexisting advanced glycation end products is associated with reduced renal fibrosis in experimental diabetes. FASEB J, 2003 Sep,17(12):1762-1764
13 Oldfield MD, Bach LA, Forbes JM, et al. Advanced glycation end products cause epithelial-myofibroblast transdifferentiation via the receptor for advanced glycation end products (RAGE). J Clin Invest, 2001 Dec, 108(12):1853-1863
14 Cisarik-Fredenburg P. Discoveries in research on diabetic keratopathy[J]. Optometry. 2001,72(11):691-704
15 Barile GR, Pachydaki SI, Tari SR, et al. The RAGE axis in early diabetic retinopathy[J].Invest Ophthalmol Vis Sci,2005, 46(8):2916-2924
16 Hudson BI, Stickland MH, Futers TS, et al. Effects of novel polymorphisms in the RAGE gene on transcriptional regulation and their association with diabetic retinopathy[J].Diabetes, 2001,50(6):1505-1511
17 Kislinger T, Fu C, Huber B, et al. N(epsilon)-(carboxymethyl)lysine adducts of proteins are ligands for receptor for advanced glycation end products that activate cell signaling pathways and modulate gene expression[J]. J Biol Chem,1999,274(44):31740-31749
18 Yan SD, Schmidt AM, Anderson GM, et al. Enhanced cellular oxidant stress by the interaction of advanced glycation end products with their receptor/binding proteins [J]. J Biol Chem,1994,269(13):9889-9897
19 Williamson JR, Chang K, Frangos M, et al. Hyperglycemic pseudo-hypoxia and diabetic complications[J]. Diabetes. 1993,42(6): 801-813
20 Rubinstein MP, Parrish ST, Vernon SA. Corneal epithelial oxygen uptake rate in diabetes mellitus[J]. Eye. 1990,4 ( Pt 5):757-759
21 Ramamurthi S, Rahman MQ, Dutton GN, et al. Pathogenesis, clinical features and management of recurrent corneal erosions[J]. Eye. 2005 Jul ,15:787-789
22 Romeo G, Liu WH, Asnaghi V, et al. Activation of nuclear factor-KB induced by diabetes and high glucose regulates a proapoptotic program in retinal pericytes. Diabetes, 2002,51(7):2241-2248
23 Mizutani M, Kern TS, Lorenzi M. Accelerated death of retinal microvascular cells in human and experimental diabetic retinopathy. J Clin Invest, 1996,97:2883-2890
24 Chen ZT, Li SL, Cai EQ, et al. LPS induces pulmonary in travaseular macrophage Producing inflammatory mediators via activating NF-KB[J]. J Cell Biochem, 2003,89: 1206-1214
25 Baeuerle PA, Baltimore D: NF-kappa B: ten years after. Cell. 1996,87:13-20
26 Baeuerle PA, Henkel T: Function and activation of NF-kappa B in the immune system. Annu Rev Immunol .1994,12:141-179
27 Karin M, Liu Z, Zandi E: AP-1 function and regulation. Curr Opin Cell Biol. 1997,9:240-246
28 Demircan N, Safran BG, Soylu M, et al. Determination of vitreous interleukin-1(IL-1) and tumour necrosis factor(TNF) levels in proliferative diabetic retinopathy. Eye, 2005, 11(11): 691-704
29 Doganay S, Evereklioglu C, Er H, et al. Comparison of serum NO, TNF-alpha, IL-1beta,SIL-2R,IL-6 and IL-8 levels with grades of retinopathy in patients with diabetes mellitus. Eye,2002,16(2):163-170
30 Slepova OS, Gerasimenko VL, Zakharova GIu, et al. Comparative study of the role of cytokines in various eye diseases and Diabetic retinopathy. Vestn Oftalmol, 2001, 117(3):35-37
31 Alves M, Calegari VC, Cunha DA, et al. Increased expression of advanced glycation end-products and their receptor, and activation of nuclear factor kappa-B inlacrimal glands of diabetic rats[J]. Diabetologia.2005, 48(12):2675-2681
32 Lusis AJ. Atherosclerosis. Nature, 2000, 407:233-241
33 任昊,侯凡凡,张训.肝素对血液透析病人单核细胞 AGE受体的影响.中华内科杂志,2002,41(1):43-46
34 Verzijl N, Degroot J, Ben ZC. Crosslinking by advanced glycation end products increases the stiffness of the collagen network in human articular cartilage; a possible mechanism through which age is a risk factor for osteoarthritis. Arthritis Rheum, 2002, 46(1):1141-1143
35 Guha M, Bai W, Nadler J, et al. Molecular mechanisms of TNF-alpha gene expression in monocytic cells via hyperglycemia-induced oxidant stress dependent and independent pathways[J]. Biol Chem, 2000, 275: 17728 -17739
36 丁力 ,丁宇. 治疗糖尿病并发症药物的开发与进展[J ] ,中国新药杂志 ,1999 ,8(10)∶667-670
37 Kowluru RA, Engerman RL,Kern TS. Abnormalities of retinal metabolism in diabetes or experimental galactosemia Ⅷ. Prevention by aminogunidine[J]. Curr Eye Res, 2000, 21(4):814-819
38 Kern TS, Tang J, Mizutani M, et al. Response of capacity cell death to aminoguanidine predicts the development of retinopathy: comparison of diabetes and galactosemia[J]. Invest Ophthalmol Vis Sci,2000, 41(12):3972-3978
39 Stitt A, Gardiner TA, Alderson NL, et al. The AGE inhibitorpyridoxamine inhibits development of retinopathy in experimental diabetes[J]. Diabetes,2002, 51(9):2826-2832
40 Vasan S, Foiles PG, Founds HW. Therapeutic potential of AGE inhibitors and breakers of AGE protein cross-links [J]. Expert Opin Investing Drugs,2001, 10(11):1977-1987
41 Singh R, Barden A, Mori T, et al. Advanced glycation end-products: a review[J]. Diabetologia, 2001;44(2):129-46
42 程梅芬, 肖 洁, 周丽诺, 杨秀芳, 王纹琴. 蛋白体外非酶糖化及氨基胍、二甲双胍的抑制作用[J ]. 上海医科大学学报 1998, 25: 35
43 程梅芬,肖洁.蛋白体外非酶糖化及氨基胍,二甲双胍的抑制作用[J].上海医科大学学报,1998,25(1):35-38
44 Kumari K, Umar S, Bansal V, et al. Inhibition of diabetes-associated complications by nucleophilic compounds[J]. Diabetes, 1991,40(8):1079-1084
45 Wolff DJ, Lubeskie A. Aminoguanidine is an isoform-selective, mechanism-based inactivator of nitric oxide synthase[J]. Arch Biochem Biophys,1995,31(1):290-310
46 Brownlee M, Vlassara H, Kooney A, et al. Aminoguanidine prevents diabetes-induced arterial wall proteincross-linking[J]. Science,1986,232(4758):1629-1632
47 Azal O, Yonem A, Guler S, et al. Effects of aminoguanidine and tolrestat on the development of ocular and renal structural changes in experimental diabetic rats[J]. Diabetes Obes Metab,2002,4(1);75-79
48 Roufail E, Soulis T, Boel E, et al. Depletion of nitric oxidesynthase-containing neurons in the diabetic retina: reversal by aminoguanidine[J]. Diabetologia, 1998, 41(12): 1419-1425
49 Kihara M, Schmelzer JD, Poduslo JF, et al. Aminoguanidine effects on nerve blood flow, vascular permeability, electrophysiology, and oxygen free radicals[J]. Proc Natl Acad Sci USA,1991,88(14):6107-6111
50 Swamy-Mruthinti S, Green K, Abraham EC. Inhibition of cataracts in moderately diabetic rats by aminoguanidine[J]. Exp Eye Res,1996,62(5):505-510
51 Huijberts MS, Wolffenbuttel BH, Boudier HA, et al. Aminoguanidine treatment increases elasticity and decreases fluid filtration of large arteries from diabetic rats[J]. J Clin Invest,1993,92(3):1407-1411
52 Miyauchi Y, Shikama H, Takasu T, et al. Slowing of peripheral motor nerve conduction was ameliorated by aminogguanidine in streptozocin-induced diabetic rats[J]. Eur J Endocrinol,1996,134(4):467-473
53 应素芬,洪丽珍,张瑾等.氨基胍和导升明联合用药对糖尿病视网膜病变患者 AGEs、NO 和视力的影响[J].专题研究,2006,35(3):46-48
54 Hammes HP, Martin S, Federlin K, et al. Am inoguanidine treatment inhibits the development of experimental diabetic retinopathy[J]. Proc Natl Acad Sci USA ,1991,88: 11555-11558
55 王荣荣,曲虹,周占宇. 氨基胍对糖尿病大鼠视网膜Caspase21 和 NF-KB 表达的影响[J]. 临床眼科杂志,2006,14(5):468-470
56 曹德权,陈艳平,常业恬. 氨基胍对全肝血流阻断再灌注大 鼠 肺 损 伤 的 影 响 [J]. 中 南 大 学 学 报 ( 医 学版),2006,31(1):94-96
57 Vasan S, Foiles PG, Founds HW. Therapeutic potential of AGE inhibitors and breakers of AGE protein cross-links [J]. Expert Opin Investing Drugs,2001,10(11)1977-1987
1 Nelson JD,Wright JC. Conjunctival goblet cell densities in ocular surface disease. Arch Ophthalmol,1984,102(7): 1049-1051
2 李凤鸣.中华眼科学[M].北京:人民卫生出版社.2005,1153
3 李永浩,刘祖国,吕林.糖尿病性角膜病变[J].中国实用眼科杂志,2002,22(5):323-325
4 Creuzot-Garcher C, Lafontaine PO, Gualino O, et al. Study of ocular surface involvement in diabetic patients[J]. J Fr Ophtalmol. 2005,28(6):583-588
5 Yoon KC, Im SK, Seo MS. Changes of tear film and ocular surface in diabetes mellitus[J]. Korean J Ophthalmol. 2004,18(2):168-174
6 Seifart U, Strempel I. The dry eye and diabetes mellitus[J]. Ophthalmologe. 1994,91(2):235-239
7 Li HY, Pang GX, Xu ZZ. Tear film function of patients with type 2 diabetes[J]. ZhongguoYi XueYuan Xue Bao.2004,26(6):682-686
8 Jin J, Chen LH, Liu XL, et al. Tear film function in non-insulin dependent diabetics[J]. ZhonghuaYan Ke Za Zhi.2003,39(1):10-13
9 Kaiserman I, Kaiserman N, Nakar S, et al. Dry eye in diabetic patients[J]. Am J Ophthalmol. 2005,139(3):498-503
10 Ohashi Y. Diabetic keratopathy[J]. Nippon Ganka GakkaiZasshi, 1997,101(2):105-110
11 Goebbels M. Tear secretion and film function in insulin dependent diabetics[J]. Br J Ophthalmol, 2000, 84: 19-21
12 Dogrus M, Katakami C, moue M. Tear function and ocular surface changes innon insulin-dependent diabetes mellitus[J]. Ophthalmology, 2001,108(3):586-592
13 Nepp J, Abela C, Polzer. Is there a correlation between the severity of diabetic retinopathy and keratoconjunctivitis sicca[J]. Cornea, 2000, 19(4):487-491
14 Molloy MP, Bilis S, Herbert BR, et al. Establishment of the human reflex tear two 2 dimensional polyacrylamide gel electrophoresis reference map new proteins of potential diagnostic value[J]. Electrophoresis,1997,18(15):2811
15 Lemmen KD, Moter H, Kirchhof B, et al. Keratopathy following vitrectomy with silicone oil injection[J]. Fortschr Ophthalmol, 1989,86(6):570-573
16 Francoise B, Pierre PJ, jean MDS, et al. Flow cytometric analysis of inflammatory markers in KCS 6-month treatment with topical cyclosporine A[J]. Invest ophthalmol vis sci, 2001,42:90
17 Rehany U, Ishii Y, Lahav M, et al. Ultrastructural changes in corneas of diabetic patients: an electron-microscopy study[J].Cornea, 2000,19(4):534-538
18 Watanabe H, Katakami C, Miyata S, et al. Corneal disorders in KKAv mouse: a type 2 diabetes model[J]. Jpn J Ophthalmol, 2002,46(2):130-139
19 Saghizadeh M, Kramerov AA, Tajbakhsh J, et al. Proteinase and growth factor alterations revealed by gene microarray analysis of human diabetic corneas[J]. Invest Ophthalmol Vis Sci. 2005,46(10):3604-3615
20 Gekka M, Miyata K, Nagai Y, et al. Corneal epithelial barrier function in diabetic patients[J]. Cornea. 2004,23(1):35-37
21 Grus FH, Augustin AJ, Evangelou NG, et al. Analysis of tear-protein patterns as a diagnostic tool for the detection of dry eyes[J]. Eur J Ophthalmol, 1998, 8(2): 90-97
22 Morishige N, Chikama T. Abnormal light scattering detected by confocal biomocroscopy at the corneal epithelial basement of subjects with type Ⅱ diabetes[J]. Diabetologia, 2001, 44(3):340
23 Kim EK, Geroski DH, Holley GP. Corneal endothelial cytoskeletal changes with aging, diabetes and after cytochalasin exposure[J]. Am J Ophthalmol, 1992, 114(3):329
24 Roszkowska AM, Tringali CG, Colosi P, et al. Corneal endothelium evaluation in type Ⅰ and type Ⅱ diabetes mellitus[J]. Ophthalmologia, 1999,213(4):258
25 Inoue K, Kato S, Inoue Y, et al. The corneal endothelium and thickness in type Ⅱ diabetes mellitus[J]. Cornea, 2002,46(1):65
26 Rosenberg ME, Tervo TM, Immonen, et al. Corneal structure and sensitivity in type Ⅰdiabetes mellitus[J].Invest Ophthalmol Vis Sci, 2000, 41(10):2915
27 Saito J, Enoki M, Hara M, et al. Correlation of corneal sensation, but not of basal or reflex tear secretion, with the stage of diabetic retinopathy[J]. Cornea, 2003,22(1):15-18
28 Xu KP, Yagi Y, Toda Ⅰ, et al. Tear function index[J]. A new measure of dry eye Arch Ophthalmol, 1995,113(1):84-89
29 Martin G. Tear secretion and tear film function in insulin-dependent diabetic[J]. Br J Ophthalmol, 2000,84:19
30 黄建艳等.糖尿病性眼表病变的初步实验及临床研究.万方硕博论文全文数据库,2003
31 张汗水,周祖廉.泪腺病学.金盾出版社.1992 年 4 月第一版:10
32 Rolando M,Zierhut M. The ocular surface and tear film and their dysfunction In dry eye disease. Surv Ophthalmol,2001,45(3):203-210
33 Sato E, Mori F, Igarashi S, et al. Corneal advanced glycation end products increase in patients with proliferative diabetic retinopathy [J]. Diabetes Care,2001,24(3):479-482
34 刘阳等.晚期糖基化终产物诱导单核细胞产生炎性细胞因子.万方硕博论文全文数据库,2004
35 Beppu H, Nakamura M, Katakami C,et al.Expression of Mn-SOD mRNA in streptozotocin-induced diabetic rat cornea by in situ hybridization[J]. Nippon Ganka Gakkai Zasshi,1996,100(8): 599-604
36 Yucel I, Yucel G, Akar Y, et al. Transmission electron microscopy and autofluorescence findings in the cornea ofdiabetic rats treated with aminoguanidine[J]. Can J Ophthalmol,2006,41(1):60-66
37 Cisarik-Fredenburg P. Discoveries in research on diabetic keratopathy[J]. Optometry. 2001, 72(11): 691-704
38 Williamson JR, Chang K, Frangos M, et al. Hyperglycemic pseudo-hypoxia and diabetic complications[J]. Diabetes. 1993,42(6): 801-813
39 Rubinstein MP, Parrish ST, Vernon SA. Corneal epithelial oxygen uptake rate in diabetes mellitus[J]. Eye. 1990,4 ( Pt 5):757-759
40 Ramamurthi S, Rahman MQ, Dutton GN, et al. Pathogenesis, clinical features and management of recurrent corneal erosions[J]. Eye. 2005 Jul 15:787-789
41 Saghizadeh M, Brown DJ, Castellon R,et al. Overexpression of matrix metalloproteinase-10 and matrix metalloproteinase-3 in human diabetic corneas: a possible mechanism of basement membrane and integrin alterations[J]. Am J Pathol. 2001,158(2):723-734
42 Takahashi H, Akiba K, Noguchi T, et al. Matrix metalloproteinase activity is enhanced during corneal wound repair in high glucose condition[J]. Curr Eye Res. 2000,21(2):608-615
43 Nishizuka Y. Protein kinase C and lipid signaling for sustained cellular responses[J]. FASEB J. 1995,9(7):484-496
44 Lin N, Bazan HE. Protein kinase C subspecies in rabbit corneal epithelium: increased activity of alpha subspeciesduring wound healing[J]. Curr Eye Res. 1992,11(9):899-907
45 Troger J, Humpel C, Kremser B, et al. The effect of streptozotocin-induced diabetes mellitus on substance P and calcitonin gene-related peptide expression in the rat trigeminal ganglion[J]. Brain Res. 1999,842(1):84-91
46 Phillips A O ,Steadman R ,Morrisey K, et al. Exposure of human renal proximal tubular cells to glucose leads to accumulation of type Ⅳ collagen and fibronectin by decreased degradation [J] . Kidney Int,1997,52:973-84
47 陈百华,姜德咏,唐罗生.糖基化终产物对视网膜微血管周细胞内基质金属蛋白酶表达的影响[J].中华眼科杂志,2002,38(6):376-378
48 Baeuerle PA, Baltimore D: NF-kappa B: ten years after. Cell. 1996,87:13-20
49 Baeuerle PA, Henkel T: Function and activation of NF-kappa B in the immune system. Annu Rev Immunol . 1994,12:141-179
50 Karin M, Liu Z, Zandi E: AP-1 function and regulation. Curr Opin Cell Biol. 1997,9:240-246
51 Romeo G, Liu WH, Asnaghi V, et al. Activation of nuclear factor-KB induce by diabetes and high glucose regulates a proapoptotic program in retinal pericytes. Diabetes, 2002,51(7):2241-2248
52 Mizutani M, Kern TS, Lorenzi M. Accelerated death of retinal microvascular cells in human and experimental diabetic retinopathy. J Clin Invest, 1996, 97:2883-2890
53 Guha M, Bai W, Nadler J, et al. Molecular mechanisms of TNF-alpha gene expression in monocytic cells via hyerglyemia-induced oxidant stress dependent and independent pathways. J Biol Chem, 2000,275:17728-17739
54 Finotto S, Ohno I, Marshall JS, et al. TNF-alpha production by eosinophils in upper airways inflammation (nasal polyposis). J Immunol.1994,153:2278–2289
55 Gamache DA, Dimitrijevich SD, Weimer LK, et al. Secretion of proinflammatory cytokines by human conjunctival epithelial cells. Ocul Immunol Inflamm. 1997,5:117-128
56 Cunningham ET Jr,Stalder A,Sannap P,et al. Localization of tumor Necrosis factor receptor messenger RNA in normal and herpes simplex Virus-infected mouse eyes[J]. Invest Ophthalmol VisSei.1997,38(1):9-15
57 Chen ZT, Li SL, Cai EQ, et al. LPS induceds pulmonary in travaseular macrophages producing inflammatory mediators via activating NF-KB[J]. J Cell Biochem, 2003,89:1206-1214
58 Schwartzman ML, Stoltz RA, Rapcon MM, et al. Rapid and sterospecific activation of NF-KB by a corneal angiogenic eicosanoid in microvessel endothelial cells[J]. Invest Ophthalmol Vis Sci,1994,35:1466-1469
59 Cubitt CL, Lausch RN, Oakes JE. Differences in interleukine 26 gene expression between cultured human corneal epithelial cells and keratocytes[J]. Invest Ophthalmol Vis Sci, 1995,36:330-336
60 Jun-Suh Choi, Jeonga Kim, Dong-Hwan, et al. Faiimre to activate NF-KB promotes apoptosis of retinal ganglion cells following optic nerve transaction[J]. Brain Res, 2000, 883:60-68
61 Senftleben U, Cao Y, Xiao G, et al. Activation by IKKa of a second ,evo-lutionary conserved NF-KB signalingy[J]. Science, 2001,293(9):1497-1499
62 Green DR. Death and NF-KB in T cell activation life at the edge[J]. Mol Cell, 2003,11(3):551-552
63 Arendt CW, Albrecht B, Soos TJ, et al. Protein kinase CQ signaling from the centerof Ⅱ le T-cell synapse[J]. Curt Opin Immunol, 2002,14(3):323-330
64 Amici C, Bed G, Rossi A, et al. Activation of NF-KB kinase by herpes simplex virus type 1 A novel target for anti-herpetic therapy[J]. J Biol Chem, 2001, 276: 28759-28766
65 Yin L, Wu L, Wesche H, et al. Dofective lymphoteoxin-βreceptor-induced NF-KB transcripfional activity in NIK-deficient mice[J]. Science, 2001, 291(7): 2162-2165
66 Klinkenberg M, Van Huffel S, Heyninck K, et al. Functional redundancy of the zinc fingers of A20 for inhibition of NF-kappa B activation and protein interactions[J]. FEBS Lett, 2001,498(1):93-97
67 Fan J, Ye RD, Malik AB. Transcriptonal mechanism of acutelung injury[J]. Am J Physiol Lung Cell Mol Physiol, 2001,281(5);11037-11050
68 Ramasamy R, Vannucci SJ, Yan SS, et al. Advanced glycation end products and RAGE: a common thread in aging, diabetes, neurodegeneration, and inflammation[J]. Glycobiology, 2005,15(7):16R-28R
69 Barile GR, Pachydaki SI, Tari SR, et al. The RAGE axis in early diabetic retinopathy[J]. Invest Ophthalmol Vis Sci,2005,46(8):2916-2924
70 Hudson BI, Stickland MH, Futers TS, et al. Effects of novel polymorphisms in the RAGE gene on transcriptional regulation and their association with diabetic retinopathy[J]. Diabetes,2001,50(6):1505-1511
71 Kislinger T,Fu C,Huber B,et al. N(epsilon)-(carboxymethyl) lysine adducts of proteins are ligands for receptor for advanced glycation end products that activate cell signaling pathways and modulate gene expression[J]. J Biol Chem, 1999,274(44):31740-31749
72 Yan SD, Schmidt AM, Anderson GM, et al. Enhanced cellular oxidant stress by the interaction of advanced glycation end products with their receptors/binding proteins[J]. J Biol Chem,1994,269(13):9889-9897
73 Kislinger T, Tanji N, Wendt T, et al. Receptor for advanced glycation end products mediates inflammation and enhanced expression of tissue factor in vasculature of diabetic apolipoprotein E-null mice[J]. Arterioscler Thromb Vasc Biol,2001,21(6):905-910
74 Bucala R, Vlassara H. Advanced glycation and end products in diabetic renal and vascular disease. Am J Kindney Dis, 1995,26(6):875-888
75 Neeper M, Schmidt AM, Breet J, et al. Cloning and expression of RAGE: a cell surface receptor for advanced glycation end products of patients. J Biol Chem, 1992,267:14998-15004
76 Stitt AW, Burke GA, Chen F, et al. Advanced glycation end-product receptor interactions on microvascular cells occur within caveolin-rich membrane domains[J]. FASEB J,2000,14(15):2390-2392
77 Salahudeen AK, Kanji V, Schmidt AM, et al. Pathogenesis of diabetic nephropathy: a radical approach . Nephrol Dial Transplant,1997,12:664-668
78 Osama H, Yan SD, Schmidt AM, et al. The receptors for advanced glycation end products HAS a central role in mediating the effect of advanced glycation end products on the development of vascular disease in diabetic mellitus. Nephrol Dial Transplant,1996,11(Suppl 5):13-16
79 丁力 ,丁宇. 治疗糖尿病并发症药物的开发与进展[J ] ,中国新药杂志 ,1999 ,8(10)∶667-670
80 Kowluru RA, Engerman RL,Kern TS. Abnormalities of retinal metabolism in diabetes or experimental galactosemia Ⅷ. Prevention by aminogunidine[J]. Curr Eye Res, 2000, 21(4):814-819
81 Singh R, Barden A, Mori T, et al. Advanced glycation end-products: a review[J]. Diabetologia, 2001,44(2):129-46
82 程梅芬, 肖 洁等. 蛋白体外非酶糖化及氨基胍、二甲双胍的抑制作用[J ]. 上海医科大学学报 1998, 25: 35
83 Monnier VM. Aminoguanidine and diabetic neuropathy[J]. Eur J Endocrinol,1996,134(4):398-400
84 Tilton RG, Chang K, Hasan KS, et al. Prevention of diabetes vascular dysfunction by guanidines. Inhibition of nitric oxide synthase versus advanced glycation end-product formation[J]. Diabetes,1993,42(2):221-232
85 Miyauchi Y, Shikama H, Takasu T, et al. Slowing of peripheral motor nerve conduction was ameliorated by aminoguanidine in streptozocin-induced diabetic rats[J]. Eur J Endocrinol,1996,134(4):476-473
86 Pugliese G, Tilton RG. Williamson JR. Glucose-induced metabolic imbalances in the pathogenesis of diabetic vascular disease[J]. Diabetes Metab Rev,1991,7(1):35-59
87 Singh R, Barden A, Mori T, et al. Advanced glycation end-products: a review[J]. Diabetologia, 2001,44(2):129-46
88 Kumari K, Umar S, Bansal V, et al. Inhibition of diabetes-associated complications by nucleophilic compounds[J]. Diabetes, 1991,40(8):1079-1084
89 Corbett JA, Tilton RG, Chang K, et al. Aminoguanidine, a novel inhibitor of nitric oxide formation, prevents diabetic vascular dysfunction [J]. Diabetes,1992,41(4):552-556
90 Wolff DJ, Lubeskie A. Aminoguanidine is an isoform-selective, mechanism-based inactivator of nitric oxide synthase[J]. Arch Biochem Biophys,1995,31(1):290-310
91 Brownlee M, Vlassara H, Kooney A, et al. Aminoguanidine prevents diabetes-induced arterial wall protein cross-linking[J]. Science,1986,232(4758):1629-1632
92 Huijberts MS, Wolffenbuttel BH, Boudier HA, et al. Aminoguanidine treatment increases elasticity and decreases fluid filtration of large arteries from diabetic rats[J]. J Clin Invest,1993,92(3):1407-1411
93 Soulis-Liparota T, Cooper M, Papazoglou D, et al. Retardation by aminoguanidine of development of albuminuria, mesangial expansion, and tissue fluorescence in streptozocin-induced diabetic rat[J]. Diabetes, 1991,40(10):1328-1334
94 Soulis T, Cooper ME, Vranes D, et al. Effects of aminoguanidine in preventing experimental diabetic nephropathy are related to the duration of treatment[J]. Kidney Int,1996,50(2):627-634
95 Azal O, Yonem A, Guler S, et al. Effects of aminoguanidine and tolrestat on the development of ocular and renal structural changes in experimental diabetic rats[J]. Diabetes Obes Metab,2002,4(1):75-79
96 Roufail E, Soulis T, Boel E, et al. Depletion of nitric oxide synthase-containing neurons in the diabetic retina: reversal by aminoguanidine[J]. Diabetologia, 1998, 41(12): 1419-1425
97 Giardino I, Fard AK, Hatchell DL, et al. Aminoguanidine Inhibits reactive oxygen species formation, lipid peroxidation, and oxidant-induced apoptosis[J]. Diabetes, 1998, 47(7): 1114-1120
98 Frank RN, Amin R, Kennedy A, et al. An aldose reductase inhibitor and aminoguanidine prevent vascular endothelial growth factor expression in rats with long-term galactosemia[J]. Arch Ophthalmol, 1997,115(8):1036-1047
99 Kihara M, Schmelzer JD, Poduslo JF, et al. Aminoguanidine effects on nerve blood flow, vascular permeability, electrophysiology, and oxygen free radicals[J]. Proc Natl Acad Sci USA,1991,88(14):6107-6111
100 Swamy-Mruthinti S, Green K, Abraham EC. Inhibition of cataracts in moderately diabetic rats by aminoguanidine[J]. Exp Eye Res,1996,62(5):505-510
101 Inomata M, Hayashi M, Shumiya S, et al. Aminoguanidine-treatment results in the inhibition of lens opacification and calpain-mediated proteolysis in Shumiya cataract rats(SCR)[J]. J Biochem(Tokyo), 2000, 128(5): 771-776
102 Chen AS, Taguchi T, Sugiura M, et al. Pyridoxal-aminoguanidine adduct is more effective than aminoguanidine in preventing neuropathy and cataract in diabetic rats[J]. Horm Metab Res, 2004, 36(3) :183-187
103 Nabekura T, Koizumi Y, Nakao M, et al. Delay of cataract development in hereditary cataract UPL rats by disulfiram and aminoguanidine[J]. Exp Eye Res,2003,76(2):169-174
104 Vasan S, Foiles PG, Founds HW. Therapeutic potential of AGE inhibitors and breakers of AGE protein cross-links [J]. Expert Opin Investing Drugs,2001,10(11)1977-1987
