本研究得到了广东省临床重点重点专科（SZGSP014）和深圳市自然科学基金（JCYJ20210324125805012）的支持。

按照本协议进行的所有动物实验均在机构动物护理和使用委员会（IACUC）的批准下进行。
1.动物准备
<ol><li>准备 12 只健康的 6 周龄 SPF Wistar 雌性大鼠，体重为 160 克± 20 克。</li>
	<li>使用裂隙灯和检眼镜检查所有大鼠的眼部状况，确保没有眼前节或视网膜疾病。</li>
	<li>将所有老鼠饲养 1 周，并有足够的食物和水源。</li>
	<li>将所有大鼠随机分为正常组，东莨菪碱药物浓度2.5 mg/mL，东莨菪碱药物浓度5 mg/mL，东莨菪碱药物浓度7.5 mg/mL组，每组3只动物。</li>
</ol>2.溶液制备
<ol><li>将东莨菪碱氢溴酸盐溶解在0.9%氯化钠溶液中，制成浓度为7.5mg/mL、5mg/mL和2.5mg/mL的溶液。</li>
	<li>制备不含东莨菪碱氢溴酸盐的0.9%氯化钠溶液，用作对照组大鼠的注射剂。</li>
</ol>3.设备和材料准备
<ol><li>准备一个小动物显微镜。</li>
	<li>准备实验材料，包括 1 mL 带针头的一次性注射器 （26 G）;荧光素钠滴眼试纸;Schirmer 撕裂试纸;无水乙醇;4%多聚甲醛;二甲苯;中性香脂;苏木精、伊红;和高碘酸-希夫染色试剂盒。</li>
</ol>4.皮下注射
注意：此过程需要第二个人的帮助来帮助保护老鼠。
<ol><li>保持老鼠的身体稳定，抓住并伸展它的左（或右）后腿。 注意：助手可以帮助抱着动物。</li>
	<li>用酒精清洁注射部位。</li>
	<li>将 1 mL 带针头 （26 G） 的一次性注射器插入拇指和手指之间的皮肤褶皱底部。</li>
	<li>通过拉回注射器柱塞来吸出注射器。注射器中的任何血液都表明针头放置不当;取出并重新定位针头。</li>
	<li>以稳定的流体运动施用 0.9% 氯化钠溶液，含或不含东莨菪碱氢溴酸盐。</li>
	<li>根据不同浓度注射所有大鼠，每次注射0.5mL，每天4次（9：00,12：00,15：00和18：00），连续19天，左右肢体交替。 注意：这些组的名称如下： 不含东莨菪碱氢溴酸盐组：0组（对照组） 氢溴酸东莨菪碱组 2.5 mg/mL：2.5 组 氢溴酸东莨菪碱组 5 mg/mL：5 组 氢溴酸东莨菪碱组 7.5 mg/mL：7.5 组</li>
	<li>将动物放回笼子并监测呼吸和行为 5-10 分钟。</li>
</ol>5.泪液分泌试验（Schirmer泪液试验，STT）
<ol><li>为大鼠创建改良的滤纸条11.沿中心线（1 毫米× 15 毫米）切割一半的用于人体的滤纸条，并修剪条带的头部使其光滑。 注意：在进行泪液分泌测试之前，请手动约束大鼠的身体，以防止移动并确保暴露大鼠的眼睛。</li>
	<li>将滤纸条放在大鼠下眼睑结膜囊的外1/3处。</li>
	<li>测试时间为 5 分钟。在整个过程中控制大鼠眼睛的闭合。</li>
	<li>测量后，用镊子将滤纸条夹入微量离心管中，并在管壁上做标记，记录撕裂量。</li>
	<li>测量第 0 天、第 1 天、第 3 天、第 5 天、第 7 天、第 11 天、第 15 天和第 19 天的泪液分泌。</li>
</ol>6. 角膜荧光素染色
<ol><li>将0.5μL0.5%荧光素钠溶液滴入每只大鼠的下结膜囊中。</li>
	<li>荧光素滴注后在蓝光下观察角膜3分钟。</li>
	<li>记录每只大鼠角膜的荧光染色，观察是否存在角膜缺损。</li>
	<li>在第 0 天、第 1 天、第 3 天、第 5 天、第 7 天、第 11 天、第 15 天和第 19 天进行角膜荧光素染色。</li>
</ol>7.结膜组织的组织学观察
<ol><li>完成模型开发后，腹膜内注射0.4 mL/100 g 10%水合氯醛水溶液，对大鼠进行深度麻醉，以缓解动物的紧张。然后，通过颈椎脱位对大鼠实施安乐死。</li>
	<li>从每只大鼠的相同区域取球结膜，大小约为2mm x 2mm。</li>
	<li>立即将组织固定在4%多聚甲醛中24小时，并嵌入石蜡13中。</li>
	<li>切割 5 μm 厚的切片，并用苏木精和伊红 （HE）14 和高碘酸 （PAS） 染色（按照制造商的说明进行操作）。</li>
</ol>8.角膜和泪腺组织的组织学观察
<ol><li>完成模型开发后，按照步骤7.1中的说明对大鼠实施安乐死。</li>
	<li>取每只大鼠右侧的角膜，立即将其固定在4%多聚甲醛溶液中。</li>
	<li>沿连接耳朵和外眼角的线切开头皮和皮下组织，将切口扩大到两侧，进一步隔离淡黄色的眼眶外腺。</li>
	<li>彻底去除大鼠的皮毛，用0.9%氯化钠溶液分离眶外腺体。</li>
	<li>将分离的眶外腺体置于4%多聚甲醛溶液中24小时并包埋在石蜡中。</li>
	<li>切割~5μm厚度的连续切片，并用HE染色，用于角膜和眶外腺组织标本。</li>
</ol>9. 统计分析
<ol><li>使用适当的软件对数据进行统计分析。<ol><li>进行单因素方差分析（ANOVA）分析数据，并进行最小显著性差异（LSD）检验以进行组间比较。将统计显著性水平设置为 α = 0.05，P < 0.05 表示统计显著性。 注：SPSS 20软件用于实验数据的统计分析。</li>
	</ol></li>
</ol>

东莨菪碱诱导的大鼠泪腺功能障碍

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作者与该程序中使用的药物和材料没有潜在的利益冲突。

缺水性干眼症（ADDE）是干眼症的重要类型，约占干眼症总人数的1/317，ADDE的主要原因是泪腺病理性损伤和炎症13。对于这种类型的干眼症，最常见的临床治疗方法是人工泪液以缓解症状或局部应用类固醇或环孢菌素18，而泪腺损伤的治疗选择很少。因此，探讨泪腺功能重建对干眼症的影响，建立泪腺功能障碍动物模型非常重要。我们使用重复?...

到 2021 年，大约 12% 的人受到干眼症的严重影响1，使其成为最常见的慢性眼病之一。干眼症可分为两种类型：缺水性干眼症（ADDE）和蒸发性干眼症（EDE）2，具体取决于影响疾病的不同因素。ADDE 进一步分为干燥综合征 （SS） 和非 SS，但大多数干眼症患者在临床3 中为非 SS 患者。慢性干眼症严重影响患者的视力质量。目前，DED的常规治疗包括使用人工泪液润滑眼表和眼睑的物理治疗。然而，在许多情况下，干眼症可能无法完全治愈。因此，研究干眼症的发病机制对于开发新疗法和新药至关重要。干眼症的动物模型为进一步研究奠定了基础。
有许多方法可以构建干眼症4 的动物模型，包括通过改变激素水平来改变泪液分泌水平。例如，切除大鼠的睾丸可以减少雄激素分泌，增加泪液分泌，并降低泪液中游离分泌成分（SC）和IgA的浓度5,6。另一种方法是通过去除控制泪腺的眼表神经来指示泪腺中的自身免疫反应。此外，通过手术切除泪腺可以直接减少泪液分泌7.不断变化的环境条件也会加速泪液蒸发。例如，在低湿度和干燥通风条件下培养动物可以建立过度蒸发干眼症模型8，该模型可以与其他方法相结合，以增加干眼症的严重程度。用于诱导干眼症实验模型的主要药物是阿托品和东莨菪碱9。作为副交感神经抑制剂，两者都能诱导泪腺中胆碱能（毒蕈碱）受体的药理学阻断，并抑制泪液分泌。与注射阿托品肌肉引起的干眼症10相比，东莨菪碱对分泌腺的抑制作用更强，药物作用持续时间更长，对心脏、小肠和支气管平滑肌的作用较弱。它是干眼症动物模型最成熟的药物之一。
不同的方法可用于用东莨菪碱诱导干眼症，例如皮下注射、药物泵或贴剂应用4,11,12。为了减少对实验动物的给药频率，许多研究人员将透皮贴剂涂抹在小鼠的尾巴上或使用药物泵。但是，这两种方法都有局限性。例如，透皮贴剂的吸收需要考虑小鼠的个体吸收，这可能导致药物剂量不一致。尽管药物泵可以准确地控制每次给药的剂量，但它们并不总是与输送的药物或使用的浓度相容。它们还需要通过手术放置——这对动物的侵入性更大，需要麻醉事件，并且有可能出现手术后并发症，例如裂开。皮下注射虽然比较繁琐，但可以确保每次给药的准确剂量，并保持不同大鼠给药的一致性。同时，它成本较低，适合进行大量的动物实验。
本研究应用反复皮下注射东莨菪碱建立干眼症大鼠模型。我们分析干眼症指标，如角膜缺损、泪液分泌水平以及角膜、结膜和泪腺的病理形态。结合药物浓度、病理表现和干眼症状，进一步详细阐述了干眼症大鼠模型，为干眼症治疗及病理机制研究提供了更准确的实验数据。我们还为未来的研究人员详细描述了建模过程。

<table><tbody><tr><td>0.9% sodium chloride solution</td><td>SJZ No.4 Pharmaceutical</td><td>H13023201</td><td></td></tr><tr><td>4% paraformaldehyde</td><td>Wuhan Servicebio Technology Co., Ltd</td><td>G1113</td><td></td></tr><tr><td>Absolute ethanol</td><td>Sinopharm Chemical Reagent Co., Ltd.</td><td>10009218</td><td></td></tr><tr><td>Fluorescein sodium ophthalmic strips</td><td>Tianjin Yinuoxinkang Medical Device Tech Co., Ltd</td><td>YN-YG-I</td><td></td></tr><tr><td>Hematoxylin and eosin</td><td>Nanjing Jiancheng Bioengineering Institute</td><td>D006</td><td></td></tr><tr><td>Neutral balsam</td><td>Beijing Solarbio Science &amp;amp; Technology Co., Ltd.&amp;nbsp;</td><td>G8590</td><td></td></tr><tr><td>Paraffin</td><td>Beijing Solarbio Science &amp;amp; Technology Co., Ltd.</td><td>YA0012</td><td></td></tr><tr><td>Periodic Acid-Schiff Staining Kit</td><td>Beyotime Biotechnology</td><td>C0142S</td><td></td></tr><tr><td>Schirmer tear test strips</td><td>Tianjin Yinuoxinkang Medical Device Tech Co., Ltd</td><td>YN-LZ-I</td><td></td></tr><tr><td>Scopolamine hydrobromide</td><td>Shanghai Macklin Biochemical Co., Ltd</td><td>S860151</td><td></td></tr><tr><td>Small animal microscope</td><td>Head Biotechnology Co,. Ltd</td><td>ZM191</td><td></td></tr><tr><td>Xylene</td><td>Sinopharm Chemical Reagent Co., Ltd.</td><td>10023418</td><td></td></tr></tbody></table>

a rat dry eye model with lacrimal gland dysfunction induced by scopolamine

缺水性干眼症（ADDE）是一种干眼症，可导致泪液分泌量和质量减少。长时间的异常泪液分泌会导致眼表环境紊乱，包括角膜损伤和炎症。在严重的情况下，ADDE会导致视力丧失甚至失明。目前，干眼症的治疗仅限于滴眼液或物理疗法，只能缓解眼部不适症状，不能从根本上治愈干眼症。为了恢复干眼症中泪腺的功能，我们创建了东莨菪碱诱导的大鼠泪腺功能障碍的动物模型。通过对泪腺、角膜、结膜等因素的综合评估，我们旨在全面了解ADDE的病理变化。与目前的干眼小鼠模型相比，该ADDE动物模型包括泪腺的功能评估，为研究ADDE中的泪腺功能障碍提供了更好的平台。

Schirmer I 测试，SIT I 在实验开始后第0、3、5、7、11、15和19天测量大鼠的泪液量。实验结果表明，与对照组（0组）相比，东莨菪碱组（2.5组、5组、7.5组）泪液分泌量显著降低，差异有统计学意义（P < 0.01）。2.5组、5组和7.5组之间无统计学意义（P > 0.05）。不同组在天数方面没有显著差异（P > 0.05）（图1， 表1）。
角膜荧?...

Watch this Scientific Journal Video about A Rat Dry Eye Model with Lacrimal Gland Dysfunction Induced by Scopolamine at JoVE.com

作者聚焦：开发干眼动物模型的挑战和未来的研究方向

在这里，我们建立了泪腺功能障碍的大鼠模型，为缺水干眼症的研究提供依据。

东莨菪碱诱导泪腺功能障碍的大鼠干眼模型

Aqueous-deficient dry eye (ADDE) is a type of dry eye disease that can result in the reduction of tear secretion quantity and quality. Prolonged abnormal ...

开发干眼症动物模型的过程是一个漫长的过程。它需要在水稻的后腿中连续皮下注射东莨菪碱，以抑制水稻中泪腺的分泌。因此，实验者需要有足够的耐心在很长一段时间内定期服用药物。我们的方案是一种非常有用的方法，可以根据不同的药物浓度创建不同程度的干眼症模型。它可以使未来的研究人员最容易研究干眼症的治疗。我们的团队将继续探索泪腺再生领域，非常高兴能够从事3D重建和移植实验。我们的最终目标是为干眼症患者开发更先进、更有效的治疗方案。

a-rat-dry-eye-model-with-lacrimal-gland-dysfunction-induced

Research

JoVE Journal

Medicine

A Rat Dry Eye Model with Lacrimal Gland Dysfunction Induced by Scopolamine

1.7K 次观看.  Jinan University. 开发干眼症动物模型的过程是一个漫长的过程。它需要在水稻的后腿中连续皮下注射东莨菪碱，以抑制水稻中泪腺的分泌。因此，实验者需要有足够的耐心在很长一段时间内定期服用药物。我们的方案是一种非常有用的方法，可以根据不同的药物浓度创建不同程度的干眼症模型。它可以使未来的研究人员最容易研究干眼症的治疗。我们的团队将继续探索泪腺再生领域，?...

视频: 作者聚焦：开发干眼动物模型的挑战和未来的研究方向

Retinal Detachment Model in Rodents by Subretinal Injection of Sodium Hyaluronate

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the occurrence of subretinal hemorrhage can affect photoreceptor cell death in the detached retina. Hence, it is advantageous to create reproducible RDs without subretinal hemorrhage for evaluating photoreceptor cell death. We modified a previously reported method to create bullous and persistent RDs in a reproducible location with rare occurrence of subretinal hemorrhage. The critical step of this modified method is the creation of a self-sealing scleral incision, which can prevent leakage of sodium hyaluronate after injection into the subretinal space. To make the self-sealing scleral incision, a scleral tunnel is created, followed by scleral penetration into the choroid with a 30 G needle. Although choroidal hemorrhage may occur during this step, astriction with a surgical spear reduces the rate of choroidal hemorrhage. This method allows a more reproducible and reliable model of photoreceptor death in diseases that involve RD such as rhegmatogenous RD, retinopathy of prematurity, diabetic retinopathy, central serous chorioretinopathy, and age-related macular degeneration (AMD).

Creating experimental retinal detachments with a reproducible and sustained height of detachment, and without subretinal hemorrhage, is important for studying the pathophysiology of photoreceptor cell loss in retinal disease and evaluating potential therapeutic interventions. Here, we report such a method in detail.

视网膜脱离模型鼠害由透明质酸钠的视网膜下注射

Subretinal injection of sodium hyaluronate is a widely accepted method of inducing retinal detachment (RD). However, the height and duration of RD or the ...

科研

医学

Experimental Glaucoma Induced by Ocular Injection of Magnetic Microspheres

Progress in understanding the pathophysiology, and providing novel treatments for glaucoma is dependent on good animal models of the disease. We present here a protocol for elevating intraocular pressure (IOP) in the rat, by injecting magnetic microspheres into the anterior chamber of the eye. The use of magnetic particles allows the user to manipulate the beads into the iridocorneal angle, thus providing a very effective blockade of fluid outflow from the trabecular meshwork. This leads to long-lasting IOP rises, and eventually neuronal death in the ganglion cell layer (GCL) as well as optic nerve pathology, as seen in patients with the disease. This method is simple to perform, as it does not require machinery, specialist surgical skills, or many hours of practice to perfect. Furthermore, the pressure elevations are very robust, and reinjection of the magnetic microspheres is not usually required unlike in some other models using plastic beads. Additionally, we believe this method is suitable for adaptation for the mouse eye.

We present a method for inducing elevated intraocular pressure (IOP), by injecting magnetic microspheres into the rat eye, to model glaucoma. This leads to strong pressure rises, and extensive neuronal death. This protocol is easy to perform, does not require repeat injections, and produces stable long-lasting IOP rises.

实验诱发青光眼通过磁性微球注射眼

Progress in understanding the pathophysiology, and providing novel treatments for glaucoma is dependent on good animal models of the disease. We present ...

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

光化学诱导后部缺血性视神经病变的大鼠模型

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue

Dry eye disease is a very common condition that causes morbidity and healthcare burden and decreases the quality of life. There is a need for a suitable dry eye animal model to test novel therapeutics to treat autoimmune dry eye conditions. This protocol describes a chronic autoimmune dry eye rat model. Lewis rats were immunized with an emulsion containing lacrimal gland extract, ovalbumin, and complete Freund&#39;s adjuvant. A second immunization with the same antigens in incomplete Freund&#39;s adjuvant was administered two weeks later. These immunizations were administered subcutaneously at the base of the tail. To boost the immune response at the ocular surface and lacrimal glands, lacrimal gland extract and ovalbumin were injected into the forniceal subconjunctiva and lacrimal glands 6 weeks after the first immunization. The rats developed dry eye features, including reduced tear production, decreased tear stability, and increased corneal damage. Immune profiling by flow cytometry showed a preponderance of CD3+ effector memory T cells in the eyeball.

This report describes a method to induce chronic experimental autoimmune dry eye in Lewis rats through immunization with an emulsion of rat lacrimal gland extract, ovalbumin, and complete Freund's adjuvant, followed by the injection of lacrimal gland extract and ovalbumin into the forniceal subconjunctiva and lacrimal glands six weeks later.

慢性自身免疫性干眼鼠模型增加效应记忆T细胞在眼球组织

Dry eye disease is a very common condition that causes morbidity and healthcare burden and decreases the quality of life. There is a need for a suitable ...

免疫与感染

A Rabbit Model of Aqueous-Deficient Dry Eye Disease Induced by Concanavalin A Injection into the Lacrimal Glands: Application to Drug Efficacy Studies

Dry eye disease (DED), a multifactorial inflammatory disease of the ocular surface, affects 1 in 6 humans worldwide with staggering implications for quality of life and health care costs. The lack of informative animal models that recapitulate its key features impedes the search for new therapeutic agents for DED. Available DED animal models have limited reproducibility and efficacy. A model is presented here in which DED is induced by injecting the mitogen concanavalin A (Con A) into the orbital lacrimal glands of rabbits. Innovative aspects of this model are the use of ultrasound (US) guidance to ensure optimal and reproducible injection of Con A into the inferior lacrimal gland; injection of Con A into all orbital lacrimal glands that limits compensatory production of tears; and use of periodic repeat injections of Con A that prolong the state of DED at will. DED and its response to test agents are monitored with a panel of parameters that assess tear production, the stability of the tear film, and the status of the corneal and conjunctival mucosa. They include tear osmolarity, tear break-up time, Schirmer&#39;s tear test, rose bengal staining, and tear lactoferrin levels. The induction of DED and the monitoring of its parameters are described in detail. This model is simple, robust, reproducible, and informative. This animal model is suitable for the study of tear physiology and of the pathophysiology of DED as well as for the assessment of the efficacy and safety of candidate agents for the treatment of DED.

This article describes the development of a method to induce acute or chronic dry eye disease in rabbits by injecting concanavalin A to all portions of the orbital lacrimal gland system. This method, superior to those already reported, generates a reproducible, stable model of dry eye suitable for the study of pharmacological agents.

由康卡纳林注射到乳原区引起的水性干眼病的兔子模型:药物功效研究的应用

Dry eye disease (DED), a multifactorial inflammatory disease of the ocular surface, affects 1 in 6 humans worldwide with staggering implications for ...

Establishment of a Severe Dry Eye Model Using Complete Dacryoadenectomy in Rabbits

Dry eye disease (DED) is a complex disease with multiple etiologies and variable symptoms, having ocular surface inflammation as its key pathophysiologic step. Despite advances in our understanding of DED, significant knowledge gaps remain. Advances are limited in part due to the lack of informative animal models. The authors recently reported on a method of DED induced by injecting all orbital lacrimal gland (LG) tissues with the lectin concanavalin A. Here, we report a novel model of aqueous-deficient DED based on the surgical resection of all orbital LG (dacryoadenectomy) tissues. Both methods use rabbits because of their similarity to human eyes in terms of the size and structure of the ocular surface. One week after removal of the nictitating membrane, the orbital superior LG was surgically removed under anesthesia, followed by removal of the palpebral superior LG, and finally removal of the inferior LG. Dacryoadenectomy induced severe DED, evidenced by a marked reduction in the tear break up time test and the Schirmer&#39;s tear test, and significantly increased tear osmolarity and rose bengal staining. Dacryoadenectomy-induced DED lasted at least eight weeks. There were no complications and animals tolerated the procedure well. The technique can be mastered relatively easily by those with adequate surgical experience and appreciation of the relevant rabbit anatomy. Since this model recapitulates the features of human aqueous-deficient DED, it is suitable for studies of ocular surface homeostasis, DED, and candidate therapeutics.

A novel approach is presented to induce chronic dry eye disease in rabbits by surgically removing all orbital lacrimal glands. This method, distinct from those previously reported, produces a stable, reproducible model of aqueous deficient dry eye well suited to study tear physiology and pathophysiology and ocular therapeutics.

在兔子中使用完全的白眼切除术建立严重干眼模型

Dry eye disease (DED) is a complex disease with multiple etiologies and variable symptoms, having ocular surface inflammation as its key pathophysiologic ...

Establishment of A Mouse Model of Aqueous Deficiency Dry Eye

Dry eye disease is a prevalent condition affecting 5%-50% of the global population. Animal model investigations play a crucial role in understanding its underlying mechanisms. Therefore, we developed a mouse model of dry eye disease by surgically removing both the extraorbital lacrimal glands (ELG) and intraorbital lacrimal glands (ILG) to investigate the ocular surface pathology in the context of aqueous deficiency dry eye. Two weeks post operation, the mice exhibited severe dry eye manifestations, including reduced tear secretion, corneal epithelial irregularities, positive fluorescein sodium staining, and neovascularization. Histological examination via hematoxylin and eosin staining revealed inflammatory cell infiltration and corneal epithelium dysplasia. Immunofluorescence staining and quantitative reverse-transcription polymerase chain reaction revealed decreased expression of the normal corneal epithelial biomarkers K12 and Pax6 and increased expression of Sprr1b in the corneal epithelium. These ocular manifestations indicated abnormal corneal epithelial differentiation. Furthermore, immunofluorescence staining of Ki67 revealed the increasing cell proliferation. In conclusion, the ELG plus ILG excision model proved suitable for studying changes in the ocular surface and elucidating the mechanisms underlying aqueous deficiency dry eye.

Here, we present a method to establish a mouse model of aqueous-deficient dry eye by excising the extraorbital and infraorbital lacrimal glands and evaluate the changes in the ocular surface in aqueous deficiency dry eye.

Dry eye disease is a prevalent condition affecting 5%-50% of the global population. Animal model investigations play a crucial role in understanding its ...

Assessment of Cornea and Tear Smoothness in an Autoimmune Dry Eye Rat Model

Source: Hou, A. et al., <a href="https://app.jove.com/v/55592">A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue.</a>&#160;J. Vis. Exp. (2017)
This video demonstrates a method for assessing corneal smoothness and tear film stability in an autoimmune rat model. It highlights the impact of gland dysfunction on tear film stability, crucial for understanding dry eye disease.

自身免疫性干眼症大鼠模型中角膜和泪液光滑度的评估

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

JoVE Encyclopedia of Experiments

Immunology

Immunopathology

Assay and Analysis

Performing Injection into Orbital Lobe of Superior Lacrimal Gland: A Technique for Inducing Dry Eye Disease in Rabbit Model

Source: Honkanen, R. A. et al, <a href="https://www.jove.com/v/59631">A Rabbit Model of Aqueous-Deficient Dry Eye Disease Induced by Concanavalin A Injection into the Lacrimal Glands: Application to Drug Efficacy Studies.</a> J. Vis. Exp. (2020).
This video describes the procedure to induce dry eye disease in a rabbit model through an injection of Concanavalin A into the orbital lobe of the superior lacrimal gland. The developed model is useful to study the effectiveness of various drugs for treating dry eye disease.

泪腺上腺眶叶注射:兔模型诱导干眼症的技术

Source: Honkanen, R. A. et al, A Rabbit Model of Aqueous-Deficient Dry Eye Disease Induced by Concanavalin A Injection into the Lacrimal Glands: ...

生物学

Large Animal Models

Leporine

A Technique to Induce Chronic Experimental Autoimmune Dry Eye in Rats

Source: Hou, A., et al. <a href="https://app.jove.com/v/55592">A Chronic Autoimmune Dry Eye Rat Model with Increase in Effector Memory T Cells in Eyeball Tissue.</a> J. Vis. Exp. (2017).
This video demonstrates a technique to induce chronic experimental autoimmune dry eye disease in rats. Upon injecting an antigen-adjuvant emulsion containing autoantigens and foreign antigens into a rat at predefined intervals, chronic eye inflammation is induced in the immunized rat by injecting the same antigens into the ocular surface tissues.