11β-羟基类固醇脱氢酶Ⅰ型在不同年龄段大鼠眼内的表达与分布

The Distribution and Expression of 11β-Hydroxysteroid Dehydrogenase Type 1 in Rat Eyes of Different Ages

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中文摘要：目的：观察糖皮质激素代谢酶11β-羟基类固醇脱氢酶Ⅰ型（11β-HSD1）在不同年龄Sprague-Dawlay （SD）大鼠眼内的表达与分布。方法：实验研究。将SD大鼠按年龄分为6组：1~2日龄新生组（PND1）、5~6日龄组（PND5）、10~11日龄组（PND10）、14~15日龄刚开眼组（PND15）、20~21日龄组（PND20）和8~9周龄成年组（M），每组6只。采用免疫组织化学染色法（IHC）检测从新生、开眼到成年等不同发育阶段SD大鼠眼组织内11β-HSD1蛋白的表达和分布情况，并通过Real time RT-PCR和Western blot进一步验证11β-HSD1在不同年龄段大鼠视网膜内的表达差异。组间数据比较采用单因素方差分析或t检验分析。结果： IHC显示11β-HSD1在各年龄组SD大鼠眼角膜、虹膜、视网膜、脉络膜、晶状体、睫状体等组织中均有表达，新生SD大鼠眼组织中的11β-HSD1阳性细胞数更多，染色更明显。Real time RT-PCR结果显示，角膜内11β-HSD1 mRNA相对表达量在SD大鼠出生后10 d达到高峰，SD大鼠开眼后开始逐渐下降，差异有统计学意义（F=8.40，P<0.001）。开眼前SD大鼠视网膜内的11β-HSD1 mRNA相对表达量高于开眼后和成年，差异有统计学意义（F=83.50，P<0.001）。Western bolt进一步证实，新生大鼠视网膜内的11β-HSD1蛋白表达量高于刚开眼大鼠和成年大鼠，差异有统计学意义（t=4.76，P=0.009；t=4.24，P=0.013）。结论： 11β-HSD1在SD大鼠眼组织内广泛分布，其表达量随年龄而变化，开眼前SD大鼠眼组织尚未分化发育成熟，其表达量较高，提示11β-HSD1可能与SD大鼠眼组织的分化、发育和成熟有关。 Objective: To observe the distribution and expressive changes of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in different age’s rat eyes. Methods: In this experimental research, according to age, SD rats were divided into 6 groups with 6 subjects in each: postnatal days 1-2 (PND1) group, postnatal days 5-6 (PND5) group, postnatal days 10-11 (PND10) group, postnatal days 14-15 (PND15) group, postnatal days 20-21 (PND20) group, and 8-9 weeks mature adults rats (M) group. Immunohistochemistry (IHC) assay was used to detect the distribution of 11β-HSD1 protein in the eyes of SD rats at different developmental stages from neonatal, postnatal eyelid opening to mature adult, and the expressions of 11β-HSD1 mRNA and protein levels in the retina or cornea were further verified by real time RT-PCR and Western blot. The data were analyzed by one-way ANOVA or independent samplet-test. Results: 11β-HSD1 protein was widely expressed in such as cornea, iris, retina, choroid, lens, ciliary body of the different age’s rat eyes by IHC staining, and there was more 11β-HSD1 positive cells in neonatal rat eyes. Real time RT-PCR showed that the relative expression of mRNA of11β-HSD1 peaked at postnatal days 10 in the cornea and decrease gradually after eyelid opening of SD rats (F=8.40, P<0.001) the relative expression of11β-HSD1 mRNA in the retina of SD rats before eyelid opening was significantly higher than that of eyelid opening rats and adult rats (F=83.50, P<0.001). It was confirmed the relative expression of 11β-HSD1 protein was significantly higher in the retina of neonatal rats than that of eyelid opening rats and mature adult rats by western blot (t=4.76, P=0.009 t=4.24, P=0.013). Conclusion: 11β-HSD1 is widely distributed in SD rat eyes of different ages. The level of 11β-HSD1 expression varies in different age groups. It tends to be higher in eyes of SD rat before eyelid opening. The study suggests that 11β-HSD1 may play an important role in the differentiation, development and maturation of the SD rat eyes.

外文摘要：Objective: To observe the distribution and expressive changes of 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) in different age’s rat eyes. Methods: In this experimental research, according to age, SD rats were divided into 6 groups with 6 subjects in each: postnatal days 1-2 (PND1) group, postnatal days 5-6 (PND5) group, postnatal days 10-11 (PND10) group, postnatal days 14-15 (PND15) group, postnatal days 20-21 (PND20) group, and 8-9 weeks mature adults rats (M) group. Immunohistochemistry (IHC) assay was used to detect the distribution of 11β-HSD1 protein in the eyes of SD rats at different developmental stages from neonatal, postnatal eyelid opening to mature adult, and the expressions of 11β-HSD1 mRNA and protein levels in the retina or cornea were further verified by real time RT-PCR and Western blot. The data were analyzed by one-way ANOVA or independent samplet-test. Results: 11β-HSD1 protein was widely expressed in such as cornea, iris, retina, choroid, lens, ciliary body of the different age’s rat eyes by IHC staining, and there was more 11β-HSD1 positive cells in neonatal rat eyes. Real time RT-PCR showed that the relative expression of mRNA of11β-HSD1 peaked at postnatal days 10 in the cornea and decrease gradually after eyelid opening of SD rats (F=8.40, P<0.001) the relative expression of11β-HSD1 mRNA in the retina of SD rats before eyelid opening was significantly higher than that of eyelid opening rats and adult rats (F=83.50, P<0.001). It was confirmed the relative expression of 11β-HSD1 protein was significantly higher in the retina of neonatal rats than that of eyelid opening rats and mature adult rats by western blot (t=4.76, P=0.009 t=4.24, P=0.013). Conclusion: 11β-HSD1 is widely distributed in SD rat eyes of different ages. The level of 11β-HSD1 expression varies in different age groups. It tends to be higher in eyes of SD rat before eyelid opening. The study suggests that 11β-HSD1 may play an important role in the differentiation, development and maturation of the SD rat eyes.

外文关键词：

11β-hydroxysteroid dehydrogenase type 1glucocorticoideye tissueexpressiondevelopment

作者：

陈薪安、林永、叶菊秀、吴昔昔

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作者单位：

1温州医科大学基础医学院，温州　325035

2温州医科大学附属眼视光医院　卫生部视觉科学研究重点实验室，温州　325027

关键词：

11β-羟基类固醇脱氢酶Ⅰ型糖皮质激素眼组织表达发育

基金：

国家自然科学基金浙江省自然科学基金温州市科技项目National Natural Science Foundation of ChinaZhejiang Provincial Natural Science Foundation of ChinaProject of Wenzhou Science & Technology Bureau

项目编号：

31301070Y2100924Y200800231301070Y2100924Y2008002

出版年：

2022

DOI：

10.3760/cma.j.cn115909-20211022-00425

中华眼视光学与视觉科学杂志

中华医学会

中华眼视光学与视觉科学杂志

CSTPCDCSCD

影响因子：0.783

ISSN：1674-845X

年,卷(期)：2022.24(8)

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