性激素受体在牛眼小梁细胞中的表达

目的了解牛眼小梁组织是否会表达性激素受体。方法体外培养新生牛眼小梁细胞，传至3代。用逆转录聚合酶链反应的方法检测牛眼小梁组织中性激素受体蛋白mRNAs(AR，EPα和PR)的表达，用免疫组织化学技术来定位这些受体。结果体外培养细胞符合牛眼小梁细胞的特点。RT-PCR检测到这些受体的mRNAs，AR、ER、PR免疫组化染色结果呈阳性。结论牛眼小梁细胞有性激素受体表达，这些受体在房水排除及青光眼发生方面可能有重要的病理生理学意义。     

Lectin receptors in the human cornea

Five different biotin labeled lectins, Concanavalin-A (Con A), wheat germ agglutinin (WGA), Ricinus communis agglutinin I (RCA1), Ulex europaeus agglutinin (UEA1), and soybean agglutinin (SBA) were used to study lectin receptors on formalin-fixed paraffin embedded human corneas. Con A stained the cytoplasm, cell, and nuclear membranes of the epithelial cells and stained the stroma diffusely. WGA stained the superficial epithelial cells, the epithelial cell membranes, and the keratocytes of the stroma. SBA did not react with any of the corneal layers. RCA1 heavily stained the keratocytes but did not stain the epithelium. UEA1 lightly stained the epithelial cell cytoplasm and interstitial stroma. All staining reactions could be abolished by omission of the lectin or by the use of the appropriate inhibitory sugar. The lectin binding patterns reported here provide a means for further investigation of carbohydrate structures in the human cornea in both normal and disease states.     

Expression of sex hormone receptors and cell cycle proteins in melanocytic lesions of the ocular conjunctiva

Background Both dermal and ocular melanocytic nevi have been reported to undergo changes during pregnancy. This has been proposed to be related to hormonal influences; however, few studies have provided any proof. We therefore set out to evaluate the expression of sex hormone receptors and cell cycle proteins in melanocytic lesions of the ocular conjunctiva. Methods Formalin-fixed, paraffin-embedded material from 76 tumors - 69 conjunctival nevi, 5 specimens of primary acquired melanosis (PAM), and 2 conjunctival melanomas - were included in a tissue microarray (TMA) format. The TMA sections were analyzed by immunohistochemistry with antibodies for progesterone and estrogen receptors, and cell cycle-related proteins (p16, MIB1-Ki67). Results Progesterone receptors were highly (96%) and similarly expressed in all lesions. In addition, progesterone receptor expression showed a tendency to increase with age (p=0.06). In contrast, estrogen receptor expression was completely absent in all tumors. The cell cycle regulator p16 was expressed in 97% of the lesions. The proliferation marker MIB1-Ki67 was expressed at low levels (mean±SD: 13±14%) in 79% of the lesions. No differences of expression were found between the different lesions and nevi types. The mean age of the patients was highest in conjunctival melanoma (70±22 years), followed by PAM (60±19 years) and nevi (36±18 SD years). The different types of nevi also showed significant age dependency (junctional 25±17 years, compound 34±17 years, dermal 49±15 years). Conclusion Our findings reveal the expression of progesterone, but not estrogen, in melanocytic lesions of the ocular conjunctiva. In benign conjunctival lesions, p16 and MIB1-Ki67 expression was comparable to that in benign nevi of the skin.     

Expression of interleukin-1 receptor antagonist in human cornea

The purpose of this study was to confirm the expression of interleukin-1 receptor antagonist (IL-1 Ra) in the human cornea. Four samples of human ex vivo corneal epithelium were obtained from patients undergoing photorefractive keratectomy. RT-PCR was performed using mRNA isolated from the corneal epithelium and oligo-dT primers. PCR was performed on the cDNA products using primers specific for human IL-1 Ra. The PCR products were subcloned and sequenced. Human cornea sections were prepared from eyes enucleated for choroidal melanoma. Immunocytochemistry was performed using goat anti-mouse polyclonal IL-1 Ra IgG and NL-577 conjugated donkey anti-goat IgG. IL-1 Ra mRNA was expressed in all ex vivo corneal epithelium samples as confirmed by sequencing of the PCR products. Immunofluorescence studies revealed strongest expression of IL-1 Ra in the superficial apical layer of corneal epithelium. Expression of IL-1 Ra may represent an endogenous mechanism of down-regulating the effects of epithelial- and tear-derived IL-1α and IL-1β on the intact epithelium in the unwounded cornea and stromal cells after injury.     

Sex hormone receptors in the human eye

Dissimilarities in ocular physiopathology exist between human males and females. These differences can be observed in the lacrimal and other eye-associated glands, the ocular surface, the crystalline lens, and the retinochoroid complexes. Literature on the subject revealed that because of sex steroid hormone (estrogen, progesterone, and androgen) actions, various physiological conditions, such as age, menstrual cycles, pregnancy, and menopause or andropause, where the hormone milieu changes, affect vision. Well-designed scientific studies are lacking on the subject, although such studies hold much potential value. This review analyzes the relatively new area of hormones and vision.     

性激素与眼表及角膜相关疾病的研究进展

性别差异是当今流行病学研究中的常见现象之一。这种现象的发生,除了与男女基因表达差异有关以外,更与性激素的调控有着密不可分的联系。在已知的各类眼科疾病中也存在着性别差异的现象。随着生物学相关研究的进展,研究人员在包括眼表及角膜在内的眼部组织中发现了越来越多的性激素受体,暗示性激素对眼表及角膜正常功能的维持有着十分重要的作用。在此,我们将对目前已知的性激素和眼表及角膜间的关系的相关报道进行综述和讨论,并对未来的相关研究进行展望。     

正常角膜缘组织中与血管生成相关的生长因子及其受体的表达

目的：探查正常人角膜缘组织里与血管生成相关的生长因子及其受体的分布。方法：用冰冻切片和链亲合素过氧化物酶染色系统，选择一组兔抗人多克隆抗体，即：血管内皮生长因子（VEGF），碱性成纤维细胞生长因子（bFGF），血小板源性生长因子（PDGF－A）和转化生长因子－β1（TGF－β1）及其受体对11例角膜移植供体材料的角膜缘组织进行了检查。结果：VEGF，bFGF,PDGF-A和TGF－β14种生长因子仅在1或2例标本中不表达外，在大部分标本中表达阳性。4种生长因子的受体在角膜缘上皮，基底膜，上皮下组织和角膜组织内阳性表达分别是：VEGF9例，7例，10例和8例；bFGF8例，9例，9例和8例，PDGF5例，4例，10例和7例；TGF－β13例，0例，11例和6例。结论：在正常角膜缘组织中，存在VEGF，bFGF ,PDGF-A和TGF－β1及其受体，可能在维持正常的角膜功能和调节角膜缘组织的生长和增殖中起着重要作用。     

Biochemical analysis of the cornea stored in steroid medium

We wished to measure changes in the tissue mass and the deoxyribonucleic acid and protein content of guinea pig corneas stored for up to 21 days in tissue culture medium with or without 1 microM hydrocortisone. Full-thickness discs 4 mm in diameter were cut from the corneas for measurement of the three variables. Whether the disc came from a steroid-treated cornea or not, and whether it came from a fresh cornea, hydrocortisone did not appear to have any adverse effect on the features studied.     

Expression of the interleukin 1 receptor antagonist in the normal human cornea

The human cornea has been shown to express a number of inflammatory cytokines including IL-1, IL-6 and IL-8. In view of the potent proinflammatory activities of interleukin-1 (IL-1), regulatory mechanisms should be present in the human cornea to control IL-1 mediated inflammatory and immune responses. This is important for the maintenance of the integrity and transparency of the cornea. To test this hypothesis, the authors determined the presence of IL-1 receptor antagonist (IL-tra) in the normal human cornea using an enzyme-linked immunosorbent assay (ELISA). IL-tra is a natural antagonist of IL-1 and competes with IL-1 for the binding to its receptors thereby blocking the inflammatory response. Corneas were either tested immediately or after a 24-hour culture period. Furthermore, the authors separately analyzed the three layers of the cornea. Their results present evidence for the constitutive expression of the IL-tra protein in the normal human cornea and show that both epithelial and stromal cells produce IL-1ra. The epithelial cells are the major source of corneal IL-1ra immunoreactivity, and secrete IL-1ra during culture. Stromal cells contain detectable, albeit low amounts of cell associated IL-1ra. No IL-1ra was detected in the endothelial cell layer. A more accurate understanding of the balance between IL-1 and IL-1ra in ocular tissues and the role of the IL-1ra under physiologie and pathophysiologic conditions will be necessary for an eventual use of IL-1 receptor antagonist as a therapeutical tool.     

Expression of the interleukin 1 receptor antagonist in the normal human cornea

The human cornea has been shown to express a number of inflammatory cytokines including IL-1, IL-6 and IL-8. In view of the potent proinflammatory activities of interleukin-1 (IL-1), regulatory mechanisms should be present in the human cornea to control IL-1 mediated inflammatory and immune responses. This is important for the maintenance of the integrity and transparency of the cornea. To test this hypothesis, the authors determined the presence of IL-1 receptor antagonist (IL-tra) in the normal human cornea using an enzyme-linked immunosorbent assay (ELISA). IL-tra is a natural antagonist of IL-1 and competes with IL-1 for the binding to its receptors thereby blocking the inflammatory response. Corneas were either tested immediately or after a 24-hour culture period. Furthermore, the authors separately analyzed the three layers of the cornea. Their results present evidence for the constitutive expression of the IL-tra protein in the normal human cornea and show that both epithelial and stromal cells produce IL-1ra. The epithelial cells are the major source of corneal IL-1ra immunoreactivity, and secrete IL-1ra during culture. Stromal cells contain detectable, albeit low amounts of cell associated IL-1ra. No IL-1ra was detected in the endothelial cell layer. A more accurate understanding of the balance between IL-1 and IL-1ra in ocular tissues and the role of the IL-1ra under physiologie and pathophysiologic conditions will be necessary for an eventual use of IL-1 receptor antagonist as a therapeutical tool     

Localization of dopamine receptors in the rabbit cornea

PURPOSE: To analyze the pharmacologic profile and the anatomic localization of the dopamine D1 and D2 receptors in sections of the rabbit cornea in normal conditions. METHODS: Samples of rabbit cornea were drawn. Biochemical and autoradiographic techniques were used on frozen sections. [3H]SCH-23390 was used as a ligand of dopamine D1 receptors and [3H]spiroperidol as a ligand of dopamine D2 receptors. RESULTS: [3H]SCH-23390 and [3H]spiroperidol were bound by sections of the rabbit cornea. The pharmacologic profile of the binding was consistent with the labeling of D1 and D2 receptors, respectively. Light microscopic analysis of the localization of D1 and D2 receptors revealed an accumulation of the two radioligands in the epithelial and in the endothelial layers of the cornea. CONCLUSION: A possible role of the dopaminergic system in the control of the corneal functions is suggested.     

The rabbit cornea lacks cholinergic receptors

Cholinergic receptors were studied in membranes prepared from rabbit cornea, iris-ciliary body, and retina, using 3H-quinuclidinyl benzilate (3H-QNB) to identify muscarinic receptors and 125I-alpha-bungarotoxin (125I-BGT) to identify nicotinic receptors. Muscarinic cholinergic receptors were not found in the cornea. As a positive control, muscarinic cholinergic receptors were characterized in preparations of the iris-ciliary body. Specific binding of 3H-QNB to iris-ciliary body membrane preparations was saturable, with a Kd of 1.3 nM QNB. Specificity of the assay for muscarinic receptors was confirmed by the relative abilities of the following compounds to displace 3H-QNB: atropine greater than pilocarpine greater than hexamethonium. Nicotinic cholinergic receptors were not found in the cornea. As a positive control, nicotinic cholinergic receptors were characterized in preparations of the retina. Specific binding of 1252-BGT to retinal membrane preparations was saturable with both high and low affinity receptors (Kd values of 1.0 nM and 93 nM BGT, respectively). Specificity of the assay for nicotinic receptors was confirmed by the relative abilities of the following compounds to prevent 125I-BGT binding: curare greater than or equal to nicotine greater than hexamethonium greater than atropine. The lack of cholinergic receptors in the cornea, which has high levels of acetylcholine and related enzymes, suggests either an extraordinary use or a lack of function for acetylcholine in this tissue.     

鼠角膜TRAIL mRNA表达与角膜移植排斥反应

目的观察正常鼠眼角膜组织中“肿瘤坏死因子相关凋亡诱导配体（TRAIL）的表达分布情况。方法采用RT-PCR技术和免疫组化技术检测TRAIL，定性、定位分析TRAIL在正常BALB／c鼠角膜组织中的表达。结果TRAIL mRNA在正常BALB／c鼠角膜组织中表达，免疫组织化学检测显示：角膜上皮层、内皮层强表达，闻质层弱表达。结论TRAIL广泛存在于角膜组织中，有抑制或减轻角膜移植免疫排斥反应的作用，但其机制有待进一步深入研究。