视黄酸在早期形觉剥夺性近视豚鼠后巩膜中的变化

目的检测豚鼠早期形觉剥夺性近视（FDM）眼巩膜中视黄酸（RA）的水平,探讨RA在FDM眼后极部巩膜中的作用。方法3周龄三色豚鼠30只随机分组,正常对照组6只,形觉剥夺15d组和形觉剥夺24d组各12只,左眼用气球头套分别遮盖15d和24d,右眼为自身对照。实验前后均测量屈光度,实验结束后摘除眼球,测量眼轴长,应用高效液相色谱法（HPLC）测定后极部巩膜RA含量。结果形觉剥夺15d组、形觉剥夺24d组动物实验眼诱导出明显的相对近视,且与正常组比较差异有统计学意义（F=23.053,P〈0.05）。形觉剥夺15d组、形觉剥夺24d组实验眼眼轴均长于自身对照眼（P〈0.05）。形觉剥夺15d组、形觉剥夺24d组实验眼巩膜中RA水平较自身对照眼均明显升高（P〈0.01）,但形觉剥夺15d组、形觉剥夺24d组间比较差异无统计学意义（P=0.857）。结论形觉剥夺可诱导近视,且近视程度在早期随时间递增,形觉剥夺眼眼轴增长。豚鼠早期形觉剥夺眼后极部巩膜RA含量增多,但在15～24d时段内差异无统计学意义。     

形觉剥夺下雏鸡后极部巩膜中MMP-2 TIMP-2及RARβ表达变化

目的 分析雏鸡形觉剥夺性近视眼及形觉剥夺性近视恢复眼后极部巩膜中基质金属蛋白酶2(matrix metallopmteinaae 2,MMP-2)、组织金属蛋白酶抑制剂2(tissue inhibitor of matrix metallopro-teinase 2,TIMP-2)及视黄酸受体-β(retinoic acid receptor-β,RARβ)的水平及其相关性.方法 选用新孵出家鸡75只,半透明眼罩遮盖的方法对左眼进行形觉剥夺,分为形觉剥夺组,遮盖14d;形觉剥夺恢复组,遮盖11d后,去遮盖3d.右眼为对照眼.取直径8mm的后极部眼组织块,分离出巩膜纤维层和软骨层.RT-PCR检测MMP-2、TIMP-2及RARβ mRNA的相对含量,并将MMP-2、TIMP-2 mRNA水平分别与RARβ的mRNA水平作线性相关分析.结果 形觉剥夺14d后,纤维层中MMP-2的表达增强(P<0.01),TIMP-2的表达减弱(P<0.01);去剥夺3d后,MMP-2的表达减弱(P<0.01),而TIMP-2的表达恢复到接近对照眼的水平(P>0.05).软骨层表达没有明显变化.形觉剥夺14d后,RARβ的表达均增强,但纤维层中的表达更为强烈(P<0.01).去剥夺3d后,RARβ的表达均明显下降(P<0.01).MMP-2与RARβmRNA水平呈正相关(P<0.05,r=0.944);TIMP-2与RARβmRNA水平呈负相关(P<0.05,r=-0.863).结论 雏鸡形觉剥夺性近视眼及形觉剥夺性近视恢复眼后极部巩膜纤维层中MMP-2、TIMP-2及RARβ的水平均发生明显变化,并有相关性.RARβ参与了巩膜纤维层细胞外基质的降解过程.     

形觉剥夺性近视鸡眼巩膜改变光电镜观察

目的　观察形觉剥夺性近视鸡眼后极部巩膜纤维层、软骨层及二者交界处的改变。方法　采用半透明眼罩遮盖的方法建立鸡眼近视模型 ,2周后检测鸡眼屈光度、眼轴长、赤道径 ,并观察后极部巩膜组织的光镜、电镜改变。结果　 2周后遮盖眼的屈光度、眼轴长和赤道径较对侧未处理眼明显增大 (P <0 .0 1)。遮盖眼后极部巩膜软骨层变厚 ,纤维层变薄 ,二者交界处可见间充质细胞。纤维层胶原纤维直径变小 ,纤维间空间增大。结论　形觉剥夺刺激鸡眼巩膜纤维层向软骨层转化 ,巩膜纤维层的改变与哺乳动物巩膜的改变相似。     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

视黄酸在形觉剥夺性近视眼巩膜中作用的研究

剥夺新生动物视网膜正常成像可导致轴性近视的发生,这种近视称为形觉剥夺性近视(FDM).高度近视形成过程中的眼轴过度延长及巩膜变薄,可能是受局部视网膜信息调控的巩膜细胞外基质(ECM)主动重塑的结果.巩膜主动重塑在眼球生长发育及正视化形成中起重要作用.视黄酸(RA)及其受体在FDM形成中对巩膜基质、软骨层和成纤维细胞层及基质降解相关蛋白酶的改变具有重要调节作用.它可能是调节FDM眼球伸长的信号分子,在巩膜重塑中扮演重要角色.     

Change in the synthesis rates of ocular retinoic acid and scleral glycosaminoglycan during experimentally altered eye growth in marmosets

PURPOSE: The purpose of this study was to examine the possibility that all-trans-retinoic acid (RA) in the eye is a signal related to changes in scleral extracellular matrix in a primate model of postnatal eye growth. METHODS: Juvenile marmosets (Callithrix jacchus) were divided into two experimental groups based on their response to monocular deprivation with diffusers: group 1, treated eyes becoming longer than fellow control eyes (n = 8), and group 2, treated eyes becoming shorter than control eyes (n = 7). Eyes were enucleated, dissected, and assayed for changes in the rates of scleral glycosaminoglycan (GAG) synthesis and ocular RA synthesis. The rate of incorporation of (35)SO4 into CPC-precipitable GAG in scleras was taken as a measure of the rate of synthesis of proteoglycans. In the same eyes the rate of RA synthesis in vivo was measured separately in the retina and the choroid/RPE (choroid with RPE attached) by HPLC. The effect of RA on the rate of scleral GAG synthesis was also examined in tissue-cultured pieces of sclera from additional marmosets. RESULTS: Induced changes in vitreous chamber length in diffuser-treated eyes correlated inversely with the rate of scleral GAG synthesis (P < 0.05) and directly correlated with the rate of RA synthesis measured separately in the retina (P < 0.05) and the choroid/RPE (P < 0.05). In group 1, the rate of scleral GAG synthesis was significantly lower (P < 0.01) in the treated eyes relative to control eyes, and the rate of RA synthesis in both the retina and the choroid/RPE was significantly higher (P < 0.01). In group 2, the rates of scleral GAG synthesis and RA synthesis in either the retina or choroid/RPE were not found to change significantly in the treated eyes compared with the control eyes. RA partially reduces the rate of scleral GAG synthesis in tissue-cultured primate sclera in a dose-dependent manner after several days. CONCLUSIONS: RA may play a role in the visual control of postnatal eye growth in primates, possibly by inducing changes in scleral extracellular matrix associated with increasing eye size. Decreasing growth rate below control levels may involve other mechanisms.     

Scleral changes in chicks with form-deprivation myopia

The sclera in myopic regions of chick eyes was studied histologically and compared to the sclera in corresponding regions of normal fellow eyes. Chicks had been monocularly deprived of form vision in the nasal half of the retina from hatching. The fellow control eye and the temporal retina of the deprived eye had normal vision. With this treatment, the resulting form-deprivation myopia and eye enlargement are restricted to the retinal region that had been form deprived. We found that the cartilaginous sclera in the myopic nasal region exhibited several differences from that in the corresponding non-myopic region: it was thicker, its cell density was lower, and the number of chondrocytes and binucleate cells was higher. In contrast, the fibrous sclera was thinner. These changes suggest that form-deprivation myopia causes an increased production of extracellular matrix and an increased level of mitotic activity in the cartilaginous sclera. As expected, the non-myopic temporal regions of experimental and control eyes did not differ in any of these parameters. The findings of the present study suggest that the eye enlargement accompanying form-deprivation myopia is not the consequence of scleral stretching but of abnormal growth.     

外源性视黄酸诱导的豚鼠近视模型的建立

目的建立豚鼠外源性视黄酸诱导近视模型,观察外源性视黄酸在三色豚鼠眼轴生长和屈光变化中的作用。方法选取体质量100g左右3～4d三色豚鼠20只,随机分为正常对照组和视黄酸诱导组。视黄酸诱导组予连续5d喂饲24mg.kg-1花生油调配的视黄酸0.5mL,正常对照组喂饲0.5mL花生油,喂饲前和喂饲后第5天分别测量其眼轴长度、屈光度,观察其变化。用HE染色测量视黄酸诱导组豚鼠巩膜和正常对照组豚鼠巩膜的厚度变化并通过透射电镜观察巩膜成纤维细胞的变化及胶原纤维的改变。结果实验前豚鼠的屈光状态和眼轴长度,两组之间差异均无统计学意义(均为P>0.05)。实验第5天,视黄酸诱导组豚鼠眼轴增长,正常对照组豚鼠的眼轴长度为(7.628±0.009)mm,视黄酸诱导组豚鼠眼轴长度为(7.743±0.005)mm,两组比较差异具有统计学意义(P<0.01);正常对照组的豚鼠眼的屈光度为(+6.50±1.60)D,视黄酸诱导组的豚鼠眼的屈光度(+1.73±0.80)D。与正常对照组相比,视黄酸诱导组的豚鼠眼屈光度平均增加了-4.80D,屈光度差异有统计学意义(P<0.01)。后极部巩膜HE染色显示,视黄酸诱导组巩膜比对照组巩膜薄。透射电镜结果显示,视黄酸诱导组巩膜胶原纤维数量减少,直径变小。结论外源性视黄酸能稳定有效地诱导眼轴增长,形成相对性的近视漂移并直接引起巩膜重塑。     

Glycosaminoglycan synthesis in the separate layers of the chick sclera during myopic eye growth: comparison with mammals.

PURPOSE: Studies in animal models of refractive development have shown that the development of and recovery from induced myopia is associated with visually-guided changes in scleral glycosaminoglycan synthesis. The present study sought to determine whether differential patterns of scleral glycosaminoglycan synthesis are present in the fibrous scleral layer of the chick during myopia development or recovery, as has previously been reported in the mammalian sclera. METHODS: Myopia was induced in young chicks by monocular deprivation of pattern vision over 5 days. Other animals underwent monocular deprivation, then had the occluder removed and were allowed 2 days of recovery. A group of age-matched normal animals served as a control. Newly synthesised glycosaminoglycans in the scleral layers were labelled in vivo, using a [(35)S]-labelled precursor delivered intraperitoneally on the final experimental day. Incorporation of this sulphate into glycosaminoglycans of the fibrous and cartilaginous scleral layers was assessed in proteinase K digests by selective precipitation with alcian blue. RESULTS: Glycosaminoglycan synthesis in the fibrous scleral layers of myopic and recovering eyes was not significantly different to contralateral control eyes (+14 +/- 7%, p = 0.09 and -2 +/- 4%, p = 0.64 respectively). In contrast, glycosaminoglycan synthesis was significantly elevated, relative to controls, in the cartilaginous scleral layer of eyes developing myopia (+63 +/- 18%, p < 0.02), whereas in recovering eyes there was found to be a significant decrease in synthesis in the cartilaginous layer (-40 +/- 6%, p < 0.001). CONCLUSIONS: The results of the current study demonstrate that the fibrous scleral layer of the chick does not display the characteristic differential patterns of glycosaminoglycan synthesis that are found in the mammalian sclera during myopia development and recovery. However, as has previously been reported, the cartilaginous layer of the chick sclera does display differential glycosaminoglycan expression, although the direction of regulation is opposite to that found in the fibrous sclera of mammals.     

Regulation of the biphasic decline in scleral proteoglycan synthesis during the recovery from induced myopia

During the recovery from form deprivation myopia (myopic defocus), the rate of proteoglycan synthesis in the posterior sclera decreases co-incident with a deceleration of axial elongation. The choroid has been implicated in the regulation of scleral proteoglycan synthesis, possibly through the synthesis and secretion of scleral growth inhibitors. Therefore these studies were carried out to attempt to establish a causal relationship between choroidal secretion and the inhibition of scleral proteoglycan synthesis during the recovery from induced myopia. Chicks were form vision deprived for 10 days followed by a recovery period (3 h–20 days) of unrestricted vision. Sclera and choroids (5 mm punches) were isolated from control and treated eyes. The rate of proteoglycan synthesis was estimated by the incorporation of ³⁵c in cetylpyridinium chloride-precipitable glycosaminoglycans by isolated sclera of control and treated eyes. Additionally, choroids from control and treated eyes were placed in co-culture with untreated age-matched normal chick sclera for 20–24 h, after which time sclera were removed and scleral proteoglycan synthesis rates were determined. Following removal of occluders, a biphasic decline was observed in scleral proteoglycan synthesis: A rapid decline in proteoglycan synthesis (−7.6% per hr; r² = 0.923) was observed over the first 12 h of recovery, followed by a slow decline extending from 12 to 96 h (−0.3% per hr; r² = 0.735). Proteoglycan synthesis rates gradually increased to control levels over the next 96 h at a rate of +0.3% per hr. No relative proteoglycan inhibition was observed when untreated sclera were co-cultured with choroids from eyes recovering for 0–4 days, whereas co-culture of untreated sclera with choroids from eyes recovering for 5 and 8 days resulted in significant inhibition of sclera proteoglycan synthesis, relative to that of sclera co-cultured with choroids from control eyes (≈−24%, P < 0.05, paired t-test). In conclusion, recovery from induced myopia is characterized by a rapid decline in proteoglycan synthesis which occurs within the first 12 h of unrestricted vision as a well as a slower more gradual decline that occurs over the next four days. Choroidal inhibition of scleral proteoglycan synthesis in vitro occurs during the second phase of decline and is most likely related to increased choroidal permeability; whereas the rapid decline in proteoglycan synthesis that occurs during the first 12 h of recovery is regulated by an independent, yet to be identified mechanism.     

Proteoglycan synthesis by scleral chondrocytes is modulated by a vision dependent mechanism.

Proteoglycan synthesis was measured in chick sclera at the onset of form-deprivation myopia, as well as in the period immediately following removal of the occluder. Two day-old chicks were monocularly form vision deprived for periods from one to ten days and proteoglycan synthesis was determined after placing posterior scleral buttons in organ culture and measuring 35SO4 incorporation into glycosaminoglycans. Following 24 hrs of form-deprivation, proteoglycan synthesis was 33% higher in myopic eyes as compared with paired control eyes. The rate of proteoglycan synthesis further increased to levels 83% higher than controls after four days of form-deprivation and remained elevated throughout the ten day period of deprivation. Removal of the occluder after 10 days of form-deprivation resulted in a rapid drop in the rate of proteoglycan synthesis to control levels within 24 hrs. Proteoglycan synthesis was also measured in scleral chondrocytes isolated from control and myopic eyes after 10 days of form-deprivation. Proteoglycan synthesis by chondrocytes from myopic eyes did not return to control levels until 48 hrs after plating. Since the rate of proteoglycan synthesis returns to control levels more quickly during the recovery period ex vivo than when scleral chondrocytes from myopic eyes are placed in cell culture, we suggest that a mechanism is present within the eye which rapidly lowers the rate of proteoglycan synthesis in response to form vision.     

Retinoic acid signals the direction of ocular elongation in the guinea pig eye

A growing eye becomes myopic after form deprivation (FD) or compensates for the power and sign of imposed spectacle lenses. A possible mediator of the underlying growth changes is all-trans retinoic acid (RA). Eye elongation and refractive error (RE) was manipulated by raising guinea pigs with FD, or a spectacle lens worn on one eye. We found retinal-RA increased in myopic eyes with accelerated elongation and was lower in eyes with inhibited elongation. RA levels in the choroid/sclera combined mirrored these directional changes. Feeding RA (25 mg/kg) repeatedly to guinea pigs, also resulted in rapid eye elongation (up to 5 times normal), and yet the RE was not effected. In conclusion, RA may act as a signal for the direction of ocular growth.     

Visual deprivation stimulates the exchange of the fibrous sclera into the cartilaginous sclera in chicks

Form deprivation myopia in chicks is a widely accepted model to study visually-regulated postnatal ocular growth. The chick sclera has a cartilaginous layer as well as the fibrous layer found in mammals. It appears that a dynamic relationship exists between these two layers during visual deprivation-induced growth. The changes in the fibrous sclera of myopic eyes, however, have not been previously described. This investigation is focused on the comparative morphological analyses of the cartilaginous and fibrous scleral changes in myopic chick eyes. The fibrous scleral changes in the posterior segment of myopic eyes were examined in detail using light and electron microscopy, and the expression of growth factors was analysed by immunohistochemistry. In the posterior segment of myopic eyes the border between the cartilaginous and fibrous layers was indistinct because of collagen bundles of the fibrous sclera that spread into the cartilaginous sclera, whereas in control eyes the distinction was clear. Various types of transitional cells, from fibroblast-like mesenchymal cells to chondrocytes, were found in the border between the cartilaginous and fibrous layers. Collagen fibrillar diameters of the fibrous sclera in the posterior segment of myopic eyes were smaller than in control, whereas those in the equatorial segment were almost the same in myopic and control eyes although the distribution of sizes was obviously different. Thus, changes in the fibrous sclera in myopic eyes of chicks seem to be similar to scleral changes in myopic eyes of mammals. The cells in the posterior sclera of myopic eyes were more intensely immunostained for TGF-beta and IGF-II than control, whereas no immunoreaction of TGF-alpha could be detected in either control or myopic eyes. These results suggest that the structural characteristics of the posterior sclera are different from those of the anterior and equatorial segments. Undifferentiated mesenchymal cells might be concentrically distributed exclusively in the innermost layer of posterior fibrous sclera. TGF-beta and IGF-II might influence cell growth, differentiation, and migration in the exaggerated scleral growth accompanying myopia.     

Induced myopia associated with increased scleral creep in chick and tree shrew eyes

PURPOSE: To investigate the role of scleral creep in the axial elongation of chick and tree shrew eyes with induced myopia. METHODS: Form-deprivation myopia was induced with a diffusing occluder worn over one eye. Scleral samples from the posterior pole and equatorial regions of myopic, contralateral (control), and age-matched normal chick and tree shrew eyes were loaded in vitro with a force of 5 g for 20 minutes while creep extension was monitored. The elastic behavior of sclera from myopic, control, and normal chick eyes was also compared. RESULTS: In both chick and tree shrew, posterior and equatorial scleral samples from myopic eyes had significantly (P < 0.05) greater creep extensions than equivalent samples from control and normal eyes (n = 10, each group). Among individual tree shrews the difference in creep rate between the sample from the myopic eye and that from the control eye correlated with vitreous chamber elongation (r = 0.746, P < 0.05) and development of myopia (r = 0.792, P < 0.01) in the deprived eye. No such association was found in the data from chicks. The elastic properties of chick sclera were unaffected in form-deprivation myopia. CONCLUSIONS: In chick and tree shrew, form-deprivation myopia is associated with increased creep rate of posterior and equatorial sclera. In tree shrew, the correlation between increased scleral creep rate and vitreous chamber elongation in myopic eyes supports the hypothesis that induced changes in the axial length of the mammalian eye are mediated by changes in the creep properties of the sclera.