碱性成纤维细胞生长因子和胰岛素样生长因子—Ⅰ及其协同作用对？…

目的 研究碱性成纤维细胞生长因子（ｂａｓｉｃ ｆｉｂｒｏｂｌａｓｔ ｇｒｏｗｔｈ ｆａｃｔｏｒ,ｂＦＧＦ）、胰岛素样生长因子－１（ｉｎｓｕｌｉｎ－ｌｉｋｅ ｇｒｏｗｔｈ ｆａｃｔｏｒ－１,ＩＧＦ－１）及二者的共同作用对体外牛晶体上皮细胞（ｂｏｖｉｎｅ ｌｅｎｓｅｐｉｔｈｅｌｉａｌ ｃｅｌｌ,ＢＬＥＣ）增殖的影响。方法 ＢＬＥＣ原代培养,取第４例细胞种入２４孔板,加入不同浓度的ｂＦＧＦ、ＩＧＦ－１,     

表皮生长因子受体家族在翼状胬肉上皮内的异常表达

目的 了解表皮生长因子受体（ｅｐｉｄｅｒｍａｌ ｇｒｏｗｔｈ ｆａｃｔｏｒ ｒｅｃｅｐｔｏｒ,ＥＧＦＲ）家族的ＥＧＦＲ、ＥｒｂＢ２及ＥｒｂＢ３蛋白在翼状胬肉上皮内的表达。方法 用免疫荧光组织化学及Ｗｅｓｔｅｒｎ ｂｌｏｔ地１５例初发期翼状胬肉患者切除的翼状胬肉组织进行ＥＧＦＲ、ＥｒｂＢ２及ＥｒｂＢ３蛋白的检测,并与正常人结膜组织进行对照。结果 免疫荧光组织化学染色显示,在正常结膜上皮中,ＥＧＦＲ蛋     

转化生长因子-β1诱导培养的人眼小梁细胞产生弹性蛋白的实验研究

目的 了解转化生长因子－β１（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈ ｆａｃｔｏｒ－β１,ＴＧＦ－β１）诱导培养培养的人眼小梁细胞产生的弹性蛋白。探讨房水引流阻力在原发性开角型青光眼（ｐｒｉｍａｒｙ ｏｐｅｎ ａｎｇｌｅ ｇｌａｕｃｏｍａ,ＰＯＡＧ）的病因和发病机制中的应用。方法 小梁网来源于除眼部疾患外的各种原因死亡的６岁以下儿童,在２４ｈ内取才于６例（１２只眼）正常眼球。人眼小梁细胞原代和传代     

生长因子对培养人眼视网膜色素上皮细胞增殖的调控作用

目的探讨生长因子对人眼视网膜色素上皮（ｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉｕｍ，ＲＰＥ）细胞增殖的调控作用。方法建立人眼ＲＰＥ细胞培养体系，利用３Ｈ－ＴｄＲ掺入法和细胞计数观察表皮生长因子（ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）、碱性成纤维细胞生长因子（ｂａｓｉｃ－ｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ，ｂ－ＦＧＦ）、胰岛素样生长因子（ｉｎｓｕｌｉｎ－ｌｉｋｅｇｒｏｗｔｈｆａｃｔｏｒ，ＩＧＦ－Ｉ）对培养ＲＰＥ细胞的作用。结果（１）ＥＧＦ，ｂ－ＦＧＦ，ＩＧＦ－Ⅰ与对照组比较，可明显增强人眼ＲＰＥ细胞ＤＮＡ合成６．６～１２．２倍，增强能力为ＥＧＦ＞ｂ－ＦＧＦ＞ＩＧＦ－Ⅰ。（２）当生长因子联合作用时，可数倍明显增强３Ｈ－ＴｄＲ掺入，能较单一因子更有效地刺激人眼ＲＰＥ增殖。结论结果提示，生长因子的协同作用可能是增殖性视网膜病变发生的重要机理之一。     

表皮生长因子对牛眼小梁细胞c—fos基因表达的诱导

目的研究表皮生长因子（ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）对小梁细胞的作用。方法应用分子杂交技术研究ＥＧＦ对体外培养的牛眼小梁细胞ｃ－ｆｏｓ基因表达的诱导和３Ｈ胸腺核苷酸（３Ｈ－ｔｈｙｍｉｄｉｎｅｉｎｃｏｒｐａｒａｔｉｏｎ，３Ｈ－ＴｄＲ）掺入法观察细胞ＤＮＡ的合成。结果小梁细胞的３Ｈ－ＴｄＲ掺入率随着ＥＧＦ浓度不同而变化，浓度为２０～１５０ｎｇ／ｍｌ时，细胞掺入率随浓度增加升高（Ｐ＜０．０１）。与对照组相比，ＥＧＦ刺激停止生长的小梁细胞０．５小时后，ｃ－ｆｏｓ基因开始表达，１小时后达高峰，至２小时后消失。不同浓度ＥＧＦ刺激小梁细胞１小时后，ｃ－ｆｏｓ基因表达呈浓度依赖性。结论实验结果表明ＥＧＦ刺激小梁细胞增殖或进入生长状态，可能与ｃ－ｆｏｓ基因产物的信号传递作用有关。     

表皮生长因子,碱性成纤维细胞生长因子及5—氟尿嘧啶对培养？…

定量观察表皮生长因子（ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）、碱性成纤维细胞生长因子（ｂａｓｉｃｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ，ｂＦＧＦ）及５－氟尿嘧啶（５－ｆｌｕｏｒｏｕｒａｃｉｌ，５－Ｆｕ）对人视网膜色素上皮（ｒｅｔｉｎａｌｐｉｇｍｅｎｔｅｐｉｔｈｅｌｉｕｍ，ＲＰＥ）细胞迁移与增殖能力的影响，探讨ＥＧＦ，ＢＦＧＦ及５－Ｆｕ在增殖玻璃体视网膜病变（ｐｒｏｌｉｆｅｒａｔ     

碱性成纤维细胞生长因子和胰岛素样生长因子-I及其协同作用对牛晶体上皮细胞增殖的影响

目的　研究碱性成纤维细胞生长因子（ｂａｓｉｃｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ，ｂＦＧＦ）、胰岛素样生长因子Ｉ（ｉｎｓｕｌｉｎｌｉｋｅ ｇｒｏｗｔｈ ｆａｃｔｏｒＩ，ＩＧＦⅠ） 及二者的共同作用对体外牛晶体上皮细胞（ｂｏｖｉｎｅ ｌｅｎｓｅｐｉｔｈｅｌｉａｌｃｅｌｌ，ＢＬＥＣ）增殖的影响。方法　ＢＬＥＣ原代培养，取第４ 代细胞种入２４ 孔板，加入不同浓度的ｂＦＧＦ、ＩＧＦⅠ、ｂＦＧＦ＋ ２０ μｇ／ＬＩＧＦⅠ，２０ ｈ 后加入３ＨＴＤＲ，１６ ｈ 后作液闪计数。结果　一定浓度的ｂＦＧＦ（１～１００ μｇ／Ｌ） 、ＩＧＦⅠ（２０～１００ μｇ／Ｌ） 在体外有促进ＢＬＥＣ 增殖的作用（ Ｐ＜ ０－０１ ） ，２０ μｇ／Ｌ的ＩＧＦⅠ可增加ｂＦＧＦ的促ＢＬＥＣ增殖作用。结论　生长因子及它们之间的协同作用在促进晶体上皮细胞异常增殖的过程中起重要作用。     

表皮生长因子、碱性成纤维细胞生长因子对人视网膜色素上皮细胞cAMP及［Ca2＋］i的影响

目前，表皮生长因子（ｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＥＧＦ）、碱性成纤维细胞生长因子（ｂａｓｉｃｆｉｂｒｏｂｌａｓｔｇｒｏｗｔｈｆａｃｔｏｒ，ｂＦＧＦ）是否以Ｃａ２＋及ｃＡＭＰ为第二信使尚存在分歧。本研究分别以Ｃａ２＋结合荧光染料（Ｆｕｒ...     

血小板源生长因子和转化生长因子—β1及其受体在视网？…

目的 观察及评估血小板源生长因子（ｐｌａｔｅｌｅｒ－ｄｅｒｉｖｅｄ ｇｒｏｗｔｈ ｆａｃｔｏｒ，ＰＤＧＦ）和转化生长因子－β１（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈ ｆａｃｔｏｒ，ＴＧＦ－β１）及其相应受体在视风原分布特征及临床意义。方法对９例孔源性视网脱离患者行玻璃体切除术同时获得２５声视网膜表面膜的分布特征及临床意义。方法 对９例孔源性视网膜脱离中层得行玻璃体切除术同时获得２５块视网膜表面膜     

人表皮生长因子对体外培养兔角膜内皮细胞影响的实验研究

目的：明确人表皮生长因子（ｈｕｍａｎ Ｅｐｉｄｅｒｍａｌ Ｇｒｏｗｔｈ Ｆａｃｔｏｒ,ｈＥＧＦ）对角膜内皮细胞的影响。方法：采用体外培养的兔角膜内皮细胞,观察接种后不同时点ｈＥＧＦ对其生长状态的影响;兔角膜片内皮损伤模型,离体培养后行Ｈ－Ｅ染色和Ｈ＾３－ＴｄＲ掺入放射自显影,观察ｈＥＧＦ对其修复的影响。结果：兔角膜内皮细胞体外培养ｈＥＧＦ组细胞数高于对照组,并使细胞形态产生纺锤形改变;角膜内皮细胞     

地塞米松体外抑制人眼小梁细胞生长及表皮生长因子mRNA的表达

目的 探讨地塞米松对培养的人眼小梁细胞生长的影响及抑制小梁细胞表达表皮生长因子（epidermal growth factor EGF)mRNA的情况。方法 取人眼小梁组织进行小梁细胞体外培养,对传3代的小梁细胞进行地塞米松处理实验。实验组在传代后的培养液中按300ug/ml加入地塞米松,另一组作为对照组进行常规培养,观察生长5d后的细胞情况,取培养7d的两组的小梁细胞分别提取RNA,用EGFcDNA探针,a-^32P同位素标记进行斑点杂交,放射自显影。对显影片用计算机激光密度扫描,测定吸光度A值相对值进行组间比较。结果 加入地塞米松300ug/ml实验组,小梁细胞生长明显受到抑制,5d时对照组细胞已经融合,地塞米松组的细胞仍呈集落状态。从对照组小梁细胞提取RNA22.5ug,地塞米松组提取RNA 14ug,取14ug两组等量RNA,用EGFcNDA探针进行斑点杂交。结果阳性。激光密度扫描值地塞米松明显低于对照组。结论 地塞米松对培养的人眼小梁细胞有明显的生长抑制作用,通过抑制总RNA转录及EGFmRNA表达而抑制小梁细胞生长。提示糖皮质激素性青光眼是因抑制了小梁细胞的多种代谢和生理功能所致。     

表皮生长因子对人视网膜色素上皮细胞Cx43mRNA表达的影响

目的:观察表皮生长因子(epidermalgrowthfactor,EGF)对培养的人视网膜色素上皮(retinapigmentepitheliumcell,RPE)细胞中缝隙连接蛋白Cx43蛋白及mRNA的表达影响。方法:不同浓度的EGF作用于培养的第4代人的RPE细胞24h,采用Westernblot的方法测定Cx43的蛋白;原位杂交观察Cx43mRNA在RPE细胞中的表达特点;RT-PCR实验对EGF影响后的RPE细胞的mRNA进行半定量观察。结果:Westernblot试验证实不同浓度的EGF均可使Cx43蛋白表达量明显减少,并且与EGF的浓度呈正相关,原位杂交显示所有细胞均表达Cx43mRNA,其表达部位主要在细胞质和核;RT-PCR实验对mRNA进行半定量观察显示表皮生长因子使RPE细胞的mRNA表达量明显减少。结论:EGF可以下调Cx43蛋白及Cx43mRNA在人RPE细胞中的表达。     

培养的人眼小梁细胞HGF和c-met的表达

目的　观察体外培养的人眼小梁细胞肝细胞生长因子 (HGF)和其受体 (c- m et)的表达。方法　成人眼小梁细胞原代培养 ,RT- PCR检测 HGF和 c- met m RNA的表达 ,免疫组织化学染色法检测 HGF和 c- met蛋白的表达 ,酶联免疫吸附法 (Elisa)检测小梁细胞分泌 HGF蛋白。结果　 RT- PCR产物为 2 6 2 bp和 30 4 bp的条带 ,为 HGF和 c- met m RNA表达阳性 ;免疫组织化学染色细胞浆呈棕色 ,为 HGF蛋白阳性表达 ,细胞膜呈棕色 ,为 c- m et蛋白阳性表达 ;Elisa法在培养的小梁细胞上清液中检测出 HGF。结论　培养的人眼小梁细胞表达 HGF和 c- m et m RNA和蛋白     

Differential effects of the EGF family of growth factors on protein secretion, MAPK activation, and intracellular calcium concentration in rat lacrimal gland

The purpose of this study was to investigate the expression of the EGF family of growth factors and EGF receptor subtypes (ErbB1-4) present in lacrimal gland and determine the effects of these growth factors on different functions of rat lacrimal gland. RT-PCR was used to detect mRNA expression in the lacrimal gland of selected members of the EGF family of growth factors, namely EGF, transforming growth factor alpha (TGF-alpha), heparin-binding EGF (HB-EGF), and heregulin. The presence of ErbB receptors was investigated by immunofluorescence microscopy and western blot analysis. The effects of EGF, TGF-alpha, HB-EGF, and heregulin on protein secretion from lacrimal gland acini were examined using a fluorescent assay for peroxidase, a marker of protein secretion. Fura-2 tetra-acetoxymethyl ester was used to measure the effects of the growth factors on intracellular [Ca2+] ([Ca2+]i) in acini. MAPK activation in acini by these growth factors was also examined by western blot analysis using antibodies specific to phosphorylated p42/44 MAPK and total p42 MAPK. Rat lacrimal gland expressed EGF, TGF-alpha, HB-EGF, and heregulin mRNA, and all four ErbB receptors were present in the lacrimal gland as detected by western blot analyses. ErbB 1 and ErbB2 were located in basal and lateral membranes of acinar and ductal cells. The location of ErbB3 could not be determined while ErbB4 was found in ductal cells. Heregulin (10(-7) m) significantly increased protein secretion in lacrimal gland acini whereas all growth factors tested significantly increased [Ca2+]i at 10(-7) m. TGF-alpha (10(-9) m), heregulin (10(-7) m), EGF (10(-7) m), and HB-EGF (10(-7) m) significantly increased the amount of phosphorylated MAPK in lacrimal gland acini. We conclude that all members of the EGF family of growth factors studied are synthesised in rat lacrimal gland, could activate all four ErbB receptors that are present in this tissue, and differentially activate lacrimal gland functions.     

CB2 cannabinoid receptors in trabecular meshwork cells mediate JWH015-induced enhancement of aqueous humor outflow facility

PURPOSE: To study the effects of JWH015, a CB2-selective agonist, on aqueous humor outflow facility, to investigate whether functional CB2 cannabinoid receptors are expressed in trabecular meshwork cells, and to study whether these receptors are involved in the enhancement of outflow facility induced by JWH015. METHODS: A porcine anterior segment perfused organ culture model was used to measure the effects of JWH015 on aqueous humor outflow facility. Immunofluorescence microscopy, Western blot analysis, and mitogen-activated protein (MAP) kinase activity assays were used to study the expression of CB2 cannabinoid receptors on cultured porcine trabecular meshwork cells and the coupling of these receptors to p42/44 MAP kinase. RESULTS: The outflow facility was increased dose dependently within 1 hour after adding 10, 30, and 100 nM of JWH015, a CB2 agonist. In addition, the effect of 100 nM of JWH015 was completely blocked by SR144528, a selective CB2 antagonist. Furthermore, the outflow-enhancing effect of JWH015 was blocked by pretreatment with PD98059, an inhibitor of the p42/44 MAP kinase pathway. In immunofluorescence microscopy and Western blot studies, positive signals were detected on cultured porcine trabecular meshwork cells with an anti-CB2 antibody. In MAP kinase assays, treatment of porcine trabecular meshwork cells with 100 nM of JWH015 activated p42/44 MAP kinase activity. Pretreatment with SR144528 blocked the effect of JWH015 on p42/44 MAP kinase activity. CONCLUSIONS: The data from this study demonstrate that the CB2-selective cannabinoid agonist JWH015 increases aqueous humor outflow facility. The results also indicate that functional CB2 cannabinoid receptors are expressed in trabecular meshwork cells, and these receptors are involved in the enhancement of outflow facility induced by JWH015.     

曲尼司特对体外培养的人眼小梁细胞转化生长因子-β2表达的抑制作用

目的　观察曲尼司特对体外培养的人眼小梁细胞转化生长因子 β2 (TGF β2 )表达的影响。方法　体外培养的第 3～ 5代人眼小梁细胞经 0 0 (对照组 )、12 5、2 5 0及 5 0 0mg/L曲尼司特处理 4 8h后 ,采用半定量逆转录聚合酶链反应 (RT PCR)观察曲尼司特对体外培养的人眼小梁细胞TGF β2 表达的影响 ,以人甘油醛 3 磷酸酯脱氢酶 (G3PDH)作为内参照。结果　 12 5、2 5 0及 5 0 0mg/L曲尼司特处理组TGF β2 /G3PDH象素值比值分别为 1 85± 0 35 ,1 6 6± 0 4 2 ,1 16± 0 2 4 ,与对照组比值 3 82± 0 5 6比较 ,差异有非常显著意义 (q′ =10 77,11 80 ,14 5 4 ;P <0 0 1) ;同时 ,TGF β2 /G3PDH比值随曲尼司特浓度增加而变小 ,呈现明显的量效关系。结论　曲尼司特能明显抑制小梁细胞TGF β2 的表达。可从发病学途径探讨曲尼司特对原发性开角型青光眼的治疗机制。