Collagen matrix perturbations in corneal stroma of Ossabaw mini pigs with type 2 diabetes.

IF 1.8 3区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Molecular Vision Pub Date : 2021-12-07 eCollection Date: 2021-01-01

Nishant R Sinha, Praveen K Balne, Filiz Bunyak, Alexandria C Hofmann, Rayne R Lim, Rajiv R Mohan, Shyam S Chaurasia

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引用次数: 0

Abstract

Purpose: Diabetes mellitus (DM) is a metabolic disorder that affects over 450 million people worldwide. DM is characterized by hyperglycemia, causing severe systemic damage to the heart, kidneys, skin, vasculature, nerves, and eye. Type 2 diabetes (T2DM) constitutes 90% of clinical cases and is the most common cause of blindness in working adults. Also, about 70% of T2DM patients show corneal complications including delayed wound healing, often described as diabetic keratopathy (DK). Despite the increasing severity of DM, the research on DK is bleak. This study investigated cellular morphology and collagen matrix alterations of the diabetic and non-diabetic corneas collected from Ossabaw mini pigs, a T2DM animal model with a "thrifty genotype."

Methods: Pig corneas were collected from six-month-old Ossabaw miniature pigs fed on a western diet (WD) for ten weeks. The tissues were processed for immunohistochemistry and analyzed using hematoxylin and eosin staining, Mason Trichrome staining, Picrosirus Red staining, Collage I staining, and TUNEL assay. mRNA was prepared to quantify fibrotic gene expression using quantitative reverse-transcriptase PCR (qRT-PCR). Transmission electron microscopy (TEM) was performed to evaluate stromal fibril arrangements to compare collagen dynamics in WD vs. standard diet (SD) fed Ossabaw pig corneas.

Results: Ossabaw mini pigs fed on a WD for 10 weeks exhibit classic symptoms of metabolic syndrome and hyperglycemia seen in T2DM patients. We observed significant disarray in cornea stromal collagen matrix in Ossabaw mini pigs fed on WD compared to the age-matched mini pigs fed on a standard chow diet using Masson Trichome and Picrosirius Red staining. Furthermore, ultrastructure evaluation using TEM showed alterations in stromal collagen fibril size and organization in diabetic corneas compared to healthy age-matched corneas. These changes were accompanied by significantly decreased levels of Collagen IV and increased expression of matrix metallopeptidase 9 in WD-fed pigs.

Conclusions: This pilot study indicates that Ossabaw mini pigs fed on WD showed collagen disarray and altered gene expression involved in wound healing, suggesting that corneal stromal collagens are vulnerable to diabetic conditions.

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患有 2 型糖尿病的 Ossabaw 迷你猪角膜基质中胶原基质的扰动。

目的：糖尿病（DM）是一种代谢性疾病，影响着全球 4.5 亿多人。糖尿病的特点是高血糖，对心脏、肾脏、皮肤、血管、神经和眼睛造成严重的全身性损害。2 型糖尿病（T2DM）占临床病例的 90%，是工作成年人失明的最常见原因。此外，约 70% 的 T2DM 患者会出现角膜并发症，包括伤口愈合延迟，通常被称为糖尿病角膜病（DK）。尽管糖尿病日益严重，但有关糖尿病角膜病的研究却很薄弱。本研究调查了从具有 "节俭基因型 "的 T2DM 动物模型奥萨博迷你猪身上采集的糖尿病和非糖尿病角膜的细胞形态和胶原基质变化：从西式饮食（WD）喂养十周的六个月大的奥萨博迷你猪身上采集猪角膜。对组织进行免疫组化处理，并使用苏木精和伊红染色法、梅森三色染色法、Picrosirus Red染色法、胶原蛋白I染色法和TUNEL法进行分析。使用透射电子显微镜（TEM）评估基质纤维排列，比较WD与标准饮食（SD）喂养的奥沙巴猪角膜中胶原蛋白的动态变化：结果：用 WD 饲喂 10 周的奥萨博迷你猪表现出 T2DM 患者典型的代谢综合征和高血糖症状。我们使用马森毛状体和毕克西里乌斯红染色法观察到，与使用标准饲料喂养的同龄迷你猪相比，使用 WD 喂养的奥萨博迷你猪角膜基质胶原基质明显混乱。此外，利用 TEM 进行的超微结构评估显示，与健康的年龄匹配角膜相比，糖尿病角膜基质胶原纤维的大小和组织发生了变化。这些变化伴随着WD喂养的猪体内胶原蛋白IV水平的显著下降和基质金属肽酶9表达的增加：这项试验性研究表明，饲喂 WD 的奥萨博迷你猪表现出胶原蛋白混乱和参与伤口愈合的基因表达改变，这表明角膜基质胶原易受糖尿病条件的影响。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Molecular Vision 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Molecular Vision is a peer-reviewed journal dedicated to the dissemination of research results in molecular biology, cell biology, and the genetics of the visual system (ocular and cortical). Molecular Vision publishes articles presenting original research that has not previously been published and comprehensive articles reviewing the current status of a particular field or topic. Submissions to Molecular Vision are subjected to rigorous peer review. Molecular Vision does NOT publish preprints. For authors, Molecular Vision provides a rapid means of communicating important results. Access to Molecular Vision is free and unrestricted, allowing the widest possible audience for your article. Digital publishing allows you to use color images freely (and without fees). Additionally, you may publish animations, sounds, or other supplementary information that clarifies or supports your article. Each of the authors of an article may also list an electronic mail address (which will be updated upon request) to give interested readers easy access to authors.