苏拉明对体外培养的视网膜色素上皮细胞增殖的影响

目的探讨苏拉明(suram in)对体外培养的猪视网膜色素上皮(retinal p igm ent ep ithelium,RPE)细胞增殖的影响。方法不同浓度苏拉明(0.05、0.1、0.2、0.4g/L)作用于RPE细胞,采用生长曲线法及MTT比色法检测苏拉明对RPE细胞增殖的影响,台盼蓝染色检测细胞活性;流式细胞仪检测细胞周期变化;透射电镜观察各浓度苏拉明作用后细胞超微结构变化。结果苏拉明对RPE细胞增殖有抑制作用(P<0.05),且呈时间剂量效应;流式细胞术检测示苏拉明将细胞阻滞于G2/M期(P<0.01);透射电镜结果示苏拉明浓度为0.05~0.2 g/L时,RPE细胞结构无明显变化,而苏拉明0.4g/L时,细胞表面微绒毛减少,染色质边集。结论苏拉明能够抑制体外培养的猪RPE细胞增殖,但随浓度增加,对细胞有潜在毒性作用。     

苏拉明对体外培养的人视网膜色素上皮细胞增殖的影响

目的 观察苏拉明(suramin)对体外培养的人视网膜色素上皮细胞(retinalpigment epithelial cells)生长、增殖的影响。方法 以300mg·L~(-1)苏拉明作用于培养的视网膜色素上皮细胞,于不同的时间点行细胞计数,绘制生长曲线,台盼蓝染色判定细胞的活性;将不同浓度的苏拉明(0、100、200、300、400mg·L~(-1))加入RPE细胞培养液,72h后用四甲基唑盐(MTT)比色法测吸光值A,并计算不同浓度条件下的细胞增殖抑制率;流式细胞技术检测细胞周期变化。结果 苏拉明在所设浓度内均抑制RPE细胞增殖,并存在时间-效应及剂量-效应关系,最大增殖抑制率约78％,72h时IC_(50)为272mg·L~(-1)。FCM示suramin使G_2/M期细胞增加了14.2％。结论 苏拉明能抑制体外培养的人视网膜色素上皮细胞增殖,可作为防治增生性玻璃体视网膜病变(proliferative vitreoretinopathy,PVR)的候选药物作进一步研究。     

曲尼司特对青光眼患者体外培养的眼部Tenon囊成纤维细胞增殖和移行的影响

目的 探讨曲尼司特(tranilast)对青光眼患者体外培养的眼部Tenon囊成纤维细胞增殖及移行的影响。方法 取青光眼患者滤过术中剪下的Tenon囊组织,在体外进行成纤维细胞原代及传代培养。分别采用MTT法、细胞计数法、免疫组化加图像分析法及划线法,研究不同浓度的曲尼司特对体外培养的成纤维细胞增殖及移行的影响及其与蛋白激酶C(PKC)表达的关系。结果 当浓度在12．5～100．0mg／L之间变化时,曲尼司特能明显抑制成纤维细胞的增殖,强度呈剂量依赖性;50．0mg／L和100．0mg／L的曲尼司特能明显抑制成纤维细胞的移行,并下调细胞内PKC的表达。结论 曲尼司特能抑制成纤维细胞的增殖及移行,这种作用可能与下调细胞内PKC的表达有关。     

血小板源性生长因子BB对体外培养兔角膜成纤维细胞增殖和细胞周期的影响

目的探讨重组人血小板源性生长因子BB(recombinant human platelet-derived growth factor-BB,rhPDGF-BB)对体外培养的兔角膜成纤维细胞增殖和细胞周期的影响。方法体外培养兔角膜成纤维细胞,分实验组和对照组,对照组弃原培养液,加无血清培养液继续培养,实验组用不同浓度的rhPDGF-BB1μg·L-1、10μg·L-1、100μg·L-1、1000μg·L-1分别作用于兔角膜成纤维细胞24h、48h、72h,用MTT比色法检测rhPDGFBB对兔角膜成纤维细胞生长的影响,流式细胞仪检测rhPDGFBB对兔角膜成纤维细胞细胞周期的影响。结果rhPDGFBB在1～100μg·L-1浓度范围内与兔角膜成纤维细胞增殖呈剂量效应关系,当浓度增大到1000μg·L-1,促增殖作用减弱。流式细胞术对细胞周期时项分析结果显示,经rhPDGFBB作用后,兔角膜成纤维细胞的DNA合成量(S%)显著升高,G1%降低,反映细胞增殖活力的增殖指数PI值增高。结论rhPDGFBB能促进兔角膜成纤维细胞的增殖和DNA合成,这一作用可能是通过促使处于G1期的兔角膜成纤维细胞进入S期来实现的。     

小檗胺抑制兔Tenon囊和滤过道内成纤维细胞增殖的实验研究

目的:研究钙调素拮抗剂小檗胺(berbamine,BER)对兔Tenon囊成纤维细胞和兔小梁切除术后滤过道内成纤维细胞增殖的抑制作用。方法:体外培养兔Tenon囊成纤维细胞,经不同浓度BER处理不同时间后,细胞计数Kit8(CCK8)法检测BER对兔Tenon囊成纤维细胞增殖的抑制作用,流式细胞仪检测其凋亡率及细胞周期的变化。HE染色检测BER对兔小梁切除术后滤过道内成纤维细胞增殖的抑制作用。结果:兔Tenon囊成纤维细胞经不同浓度BER处理不同时间后细胞增殖受到抑制,并呈时间剂量依赖性。流式细胞仪检测发现BER处理后兔Tenon囊成纤维细胞呈现G1期阻滞,细胞凋亡率明显增加,20mg/LBER处理9h,细胞凋亡率从0.64%增加到31.86%。HE染色显示BER显著抑制兔小梁切除术后滤过道内成纤维细胞的增殖。结论:小檗胺在体内外均能抑制成纤维细胞的增殖,其可能是通过诱导细胞凋亡方式抑制兔Tenon囊和滤过道内成纤维细胞的增殖。     

K115对TGF-β1诱导的人Tenon囊成纤维细胞增殖和迁移的影响及其机制

目的 探讨K115对转化生长因子-β1(TGF-β1)诱导的人Tenon囊成纤维细胞增殖和迁移的影响及其机制.方法 人Tenon囊成纤维细胞取自于河北省人民医院眼科行青光眼手术的患者.采用CCK-8实验检测K115对人Tenon囊成纤维细胞增殖的影响,筛选出TGF-β1对细胞增殖最佳作用浓度为5.0 μg·L-1,最佳...     

姜黄素和苏拉明抑制兔RPE细胞增生的对比研究

目的 对比研究姜黄素和苏拉明对兔视网膜色素上皮(RPE)细胞增生的抑制作用.以苏拉明为对照药,探讨姜黄素防治增生性玻璃体视网膜病变(PVR)的潜在价值.方法 MTT法检测不同质量浓度姜黄素和苏拉明在各时间段对体外培养的RPE细胞增生的抑制率.相关回归分析计算两种药物各时间段的半数抑制率(IC50)剂量.流式细胞仪检测姜黄素和苏拉明对RPE细胞周期及细胞凋亡的影响,并且检测姜黄素作用不同时间后RPE细胞的凋亡率.结果 姜黄素和苏拉明对RPE细胞均有明显的抑制作用,呈时间和质量浓度依赖性.姜黄素在24、48、72及96h的IC50均明显低于苏拉明.姜黄素使RPE细胞阻滞在G0/G1期,早于苏拉明的G2/M期.姜黄素可以诱导RPE细胞凋亡,苏拉明则不能.姜黄素15μg/mL作用24、48及72h后,RPE细胞的凋亡率分别为(12.83±0.13)%、(32.27±4.51)%、(56.81±8.67)%.结论 姜黄素通过诱导RPE细胞凋亡而有效抑制其增生,比苏拉明药效强且起效快,有望成为防治PVR的理想天然药物.     

曲古抑菌素A对人Tenon囊成纤维细胞作用的研究

目的：研究曲古抑菌素A( trichostatin A,TSA)对体外培养的人Tenon囊成纤维细胞( human Tenon capsule fibroblast, HTF)增殖能力以及组蛋白去乙酰化酶1( histone deacetylase 1,HDAC1)和HDAC2蛋白表达的影响。
　　方法：取青光眼滤过术中Tenon囊组织进行HTF体外培养。选取第3~6代细胞进行实验。设置空白对照组及TSA组(600nmol/L TSA加入培养液中),分别于培养后1,2,3 d应用 MTT法检测细胞活力变化;以600 nmol/L TSA处理 HTF 2 d 后, Western blot 法检测细胞中 HDAC1和HDAC2蛋白的表达。
　　结果：与对照组比较,TSA作用1d后HTF活力下降,呈时间依赖性,具有统计学差异( P<0.05)。 TSA处理HTF后2d,Western blot法检测HDAC1和HDAC2蛋白表达受到明显抑制。
　　结论：TSA可以通过抑制HDAC1和HDAC2的表达量,抑制人Tenon囊成纤维细胞增殖,从而减少术后结膜瘢痕形成可能。     

兔眼滤过术联合应用苏拉明后滤过道的组织病理学观察

目的研究小梁切除术联合应用苏拉明后滤过道的组织病理学变化。方法对20只健康家兔(40眼)均行青光眼滤过手术。随机分为两组,每组各10只(20眼)。第1组:随机选择一眼做单纯小梁切除术,另一眼做小梁切除术并辅助应用苏拉明;第2组:随机选择一眼做小梁切除术联合应用丝裂霉素(mitomycine,MMC),另一眼做小梁切除术并辅助应用苏拉明。对兔眼术后3d、7d、15d、30d、60d不同时期的滤过泡组织进行病理检查,采用HE染色和天狼星-苦味酸染色、Masson三色染色法、免疫组化方法,观察成纤维细胞、新生胶原纤维、新生血管的改变化。结果成纤维细胞:应用苏拉明组和MMC组术后滤过道成纤维细胞数目与单纯小梁切除组差异均有统计学意义(P<0.05),而该两组之间差异无统计学意义;新生胶原纤维:应用苏拉明组和MMC组术后滤过道新生胶原量与单纯小梁切除组差异均有统计学意义(P<0.05),而两组之间差异无统计学意义;新生血管:应用苏拉明组和MMC组术后滤过道成纤维细胞数目与单纯小梁切除组差异均有统计学意义(P<0.05),而两组之间差异无统计学意义。结论小梁切除术联合应用苏拉明可在术后抑制成纤维细胞增殖、新生胶原纤维的合...     

精氨酸-甘氨酸-天门冬氨酸-丝氨酸肽对人Tenon囊成纤维细胞的贴壁抑制试验观察

目的：研究精氨酸-甘氨酸-天门冬氨酸-丝氨酸（Arg-Gly-Asp-Ser，RGDS）肽对人Tenon囊成纤维细胞与纤维连接蛋白（FN）粘附、增殖的影响。方法：在传代培养的人Tenon囊成纤维细胞中加入不同浓度的RGDS（1、0.1、0.001g/L），用未贴壁细胞记数法及MTT法测定RGDS肽对人Tenon囊成纤维细胞粘附、增殖的影响。结果：RGDS肽能抑制成纤维细胞在FN上的粘附，呈浓度效应特点，能抑制成纤维细胞的增殖，呈浓度及时间效应特点。结论：在细胞水平上证实RGDS肽可阻止人Tenon囊成纤维细胞对FN的粘附和增殖，具有避免青光眼滤过术后瘢痕化形成的应用前景。     

Electrophysiological determination of the refractive state of the eye of the opossum

The refractive state of the eye of the South American opossum Didelphis marsupialis aurita was investigated with electrophysiological techniques. Using adult specimens, trapped from the wild, averaged cortical evoked responses were recorded from the region of projection of the central visual field. Stimuli consisted of a phase reversal of a square wave grating generated on a CRO screen, with luminance of 2.4 cd/m2 and contrast 0.84. The refractive state of the eye was altered by means of trial lenses and the amplitude of the cortical responses thus obtained compared to those obtained with no lens (control values). Refraction "tuning curves" were determined for each animal. The average refractive state was found to be -2.27 D indicating that this species when raised in its habitat shows, at low ambient luminosity, some degree of myopia. Determination of the Contrast Sensitivity Function indicate that induced ametropias lead to a reduction of the cut-off value of the spatial frequency and a loss of contrast sensitivity.     

Influence of exercise on the structure of the anterior chamber of the eye

Purpose

To measure changes in anterior chamber structure before and after exercise in healthy individuals using anterior segment optical coherence tomography (ASOCT).

Methods

Thirty‐two healthy young individuals performed jogging for 20 min. Eye blinking rate was recorded during rest and exercise. The anterior chamber angle (ACA), angle opening distance at 500 μm from the scleral spur (AOD500), trabecular‐iris space area at 500 μm from the scleral spur (TISA500), iris concavity (IC), iris concavity ratio (CR), iris thickness at 750 μm from the scleral spur (IT750), anterior chamber depth (ACD), anterior chamber width (ACW), pupil diameter (PD), intraocular pressure (IOP), blood pressure (BP) and heart rate (HR) were recorded before and after exercise. Anterior chamber angle (ACA), AOD500, TISA500, IC, IT750, ACD, ACW and PD were measured with ASOCT.

Results

Compared with rest, the blinking rate during exercise did not change significantly (13.04 ± 5.80 versus 13.52 ± 5.87 blinks/min, p = 0.645). The average IOP (15.4 ± 2.4 versus 12.4 ± 2.1 mmHg), ACA (35.96 ± 11.35 versus 40.25 ± 12.64 degrees), AOD500 (0.800 ± 0.348 versus 0.942 ± 0.387 mm), TISA500 (0.308 ± 0.155 versus 0.374 ± 0.193 mm²), IC (?0.078 ± 0.148 versus ?0.153 ± 0.159 mm) and CR (?0.027 ± 0.050 versus ?0.054 ± 0.056) changed significantly (all p < 0.001), while the average IT750 (0.463 ± 0.084 versus 0.465 ± 0.086 mm; p = 0.492), ACD (3.171 ± 0.229 versus 3.175 ± 0.238 mm; p = 0.543) and ACW (11.768 ± 0.377 versus 11.755 ± 0.378 mm; p = 0.122) showed no significant change after exercise.

Conclusion

The blinking rate did not change significantly during exercise, while ACA, AOD500 and TISA500 increased after exercise. Exercise also induced or increased IC. These changes in anterior chamber structure were only associated with exercise, but not with the postexercise change in PD or IOP.     

Determination of the location of the fovea on the fundus

PURPOSE: To evaluate whether the distance between optic nerve head and fovea in healthy eyes determined by scanning laser ophthalmoscope may facilitate estimation of the location of the fovea relative to the optic disc in patients with macular disease. METHODS: The angular distance was measured, in horizontal and vertical directions, between the center of the optic nerve head and the fovea in 104 eyes of 104 healthy probands. For additional evaluation of intraindividual variation in 70 of these persons the contralateral eye was measured as well. RESULTS: The distance between the optic disc and the fovea differed vertically more than horizontally (-1.5 +/- 0.9 degrees [-3.65 to +0.65 degrees ] vs. 15.5 +/- 1.1 degrees [13.0-17.9 degrees ]). There was a mean angle between the fovea and the center of the optic disc versus the horizon of -5.6 +/- 3.3 degrees. The intraindividual difference between right and left eyes was markedly lower, with average angles being 0.2 +/- 1.3 degrees vertically and 0.0 +/- 1.1 degrees horizontally. CONCLUSIONS: The distance between the optic nerve head and the fovea does not allow for a meaningful determination of the location of the fovea in eyes in which morphologic changes have occurred. The angle of rotation of the fovea relatively to the center of the optic nerve head is relatively stable. Therefore, the size of a central scotoma can be determined by movement of the blind spot according to the change of the preferred retinal locus (PRL). In addition, the knowledge of the location of the fovea enables determination of the position in the contralateral eye of the same patient.