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小鼠眶外泪腺结构、神经支配及分泌反应的年龄依赖性变化。

Age-dependent alterations in mouse exorbital lacrimal gland structure, innervation and secretory response.

作者信息

Ríos José David, Horikawa Yoshitaka, Chen Li-Li, Kublin Claire L, Hodges Robin R, Dartt Darlene A, Zoukhri Driss

机构信息

Schepens Eye Research Institute and Department of Ophthalmology, Harvard Medical School, 20 Staniford St., 02114 Boston, MA, USA.

出版信息

Exp Eye Res. 2005 Apr;80(4):477-91. doi: 10.1016/j.exer.2004.10.012.

Abstract

Several studies investigated the effect of aging on rat and human lacrimal gland physiology. However, in most of these studies, only two age groups were investigated. Furthermore, those studies did not correlate the age-related histological changes that occur in the lacrimal gland to the functional changes (nerve activity and protein secretion) that might occur with aging. Thus, the purpose of the present study was to investigate the effect of aging on lacrimal gland structure, innervation and function using BALB/c mice at different ages. Exorbital lacrimal glands were removed from 3, 8, 12, 24, and 32-month-old, male BALB/c mice, fixed, embedded and processed for histology and immunohistochemistry. Sections were stained with hematoxylin and eosin to determine morphological changes and lymphocytic infiltration; giemsa to identify mast cells; and Kinyoun's carbol fucsin solution to indicate lipofuscin-like inclusions. Parasympathetic and sympathetic nerves were identified by immunofluorescence techniques. To measure acetylcholine release and protein secretion, lacrimal gland pieces were incubated in Krebs Ringer buffer containing 5 mM KCl (control), 75 mM KCl (depolarizing buffer which activates nerves), carbachol (a cholinergic agonist, 10(-4) M), or phenylephrine (an alpha1-adrenergic agonist, 10(-4) M) for 20 min. The media were collected and analysed for acetylcholine and peroxidase using a spectrofluorometric assay. KCl-, carbachol- and phenylephrine-stimulated peroxidase secretion decreased in lacrimal glands from 8, 12, and 24-month-old mice when compared to 3-month-old animals. Both the density and distribution of parasympathetic and sympathetic nerves surrounding the acini decreased with increasing age. Acetylcholine release from lacrimal gland nerves decreased in 24-month-old mice compared to 3- and 12-month-old animals. Similarly, progressive morphological changes, including increased numbers of lipofuscin-like inclusions, mast cells and lymphocytic infiltration occurred in an age-dependent manner. We conclude that structural alterations of mouse lacrimal gland, including increased accumulation of lipofuscin-like inclusions, chronic inflammation and functional alterations including decreased acetylcholine release and protein secretion occurred with aging.

摘要

多项研究调查了衰老对大鼠和人类泪腺生理的影响。然而,在这些研究中的大多数,仅调查了两个年龄组。此外,那些研究并未将泪腺中发生的与年龄相关的组织学变化与衰老可能伴随发生的功能变化(神经活动和蛋白质分泌)联系起来。因此,本研究的目的是使用不同年龄的BALB/c小鼠来调查衰老对泪腺结构、神经支配和功能的影响。从3、8、12、24和32月龄的雄性BALB/c小鼠中取出眶外泪腺,固定、包埋并进行组织学和免疫组织化学处理。切片用苏木精和伊红染色以确定形态变化和淋巴细胞浸润;用吉姆萨染色以识别肥大细胞;用金扬氏石炭酸品红溶液显示脂褐素样包涵体。通过免疫荧光技术识别副交感神经和交感神经。为了测量乙酰胆碱释放和蛋白质分泌,将泪腺组织块在含有5 mM氯化钾(对照)、75 mM氯化钾(激活神经的去极化缓冲液)、卡巴胆碱(一种胆碱能激动剂,10⁻⁴ M)或去氧肾上腺素(一种α1-肾上腺素能激动剂,10⁻⁴ M)的克雷布斯林格缓冲液中孵育20分钟。收集培养基并使用荧光分光光度法分析乙酰胆碱和过氧化物酶。与3月龄动物相比,8、12和24月龄小鼠泪腺中氯化钾、卡巴胆碱和去氧肾上腺素刺激的过氧化物酶分泌减少。腺泡周围副交感神经和交感神经的密度和分布均随年龄增长而降低。与3月龄和12月龄动物相比,24月龄小鼠泪腺神经的乙酰胆碱释放减少。同样,包括脂褐素样包涵体数量增加、肥大细胞和淋巴细胞浸润在内的渐进性形态变化也呈年龄依赖性发生。我们得出结论,小鼠泪腺的结构改变,包括脂褐素样包涵体积累增加、慢性炎症以及功能改变,如乙酰胆碱释放和蛋白质分泌减少,均随衰老而发生。

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