In vitro effects of sex hormones in human meibomian gland epithelial cells
In vitro effects of sex hormones in human meibomian gland epithelial cells
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DOI：
10.1016/j.exer.2016.08.009
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发表时间：
2016-10-01
影响因子：
3.4
通讯作者：
Garreis, Fabian
中科院分区：
文献类型：
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作者：
Schroeder, Antje;Abrar, Daniel B.;Garreis, Fabian
睑板腺功能障碍（MGD）被认为是干眼病（DED）的最常见原因。性激素似乎在MGD的发病机制中起作用，尽管它们的参与还不完全清楚。因此，在本研究中，我们评估了二氢睾酮（DHT）和雌二醇（β-Est）对永生化人睑板腺上皮细胞系（HMGEC）的影响。Western blot检测HMGEC中性激素受体蛋白表达。进行超微结构形态学、苏丹III脂质染色、细胞增殖以及活力测定。此外，通过真实的时间RT-PCR分析用于角化（角蛋白、外皮蛋白和CK 6）、增殖（CK 5和CK 14）和脂质合成（脂肪酸合成酶和硬脂酰辅酶A去饱和酶）的MGD相关标志物的表达。Western blot显示HMGEC中存在雄激素受体（AR）、雌激素受体α和β（ER α、ER β）以及孕激素受体（PR）。在血清培养下，PR、ER α和ER β的表达显著诱导，而性激素刺激对PR、ER α和ER β的蛋白表达没有进一步的影响。我们的研究结果表明，MGD相关的性激素对HMGEC的细胞形态和脂质积累没有影响。通过应用性激素和补充钙，轻微诱导细胞增殖。然而，性激素和钙都改变了MGD相关标志物的基因表达。特别是角化基因hornerin（HRNR）和cornulin（COR）的诱导后，应用性激素和钙在无血清培养的HMGEC。这可能促进与MGD相关的角质化过程。进一步的调查是必要的，以分析（超）角化过程中发生的MGD和使用HMGEC作为体外模型。(C)2016爱思唯尔有限公司版权所有
Meibomian gland dysfunction (MGD) is considered the most common cause of dry eye disease (DED). Sex hormones seem to play a role in the pathogenesis of MGD although their involvement is not completely understood. Therefore, in the present study we evaluated the effect of dihydrotestosteron (DHT) and estradiol (beta-Est) on an immortalized human meibomian gland epithelial cell line (HMGEC). Protein expression of sex hormone receptors in HMGEC was investigated by western blot. Ultrastructural morphology, Sudan III lipid staining, cell proliferation as well as vitality assays were performed. Furthermore, expression of MGD-associated markers for keratinization (hornerin, involucrin and CK6), proliferation (CK5 and CK14) and lipid synthesis (fatty acid synthase and stearoyl-CoA desaturase) were analyzed by real time RT-PCR. Western blot revealed presence of androgen receptor (AR), estrogen receptors alpha and -beta (ER alpha, ER beta) and progesterone receptor (PR) in HMGEC. PR, ER alpha and ER beta expression was significantly induced under cultivation with serum, whereas sex hormones stimulation showed no further effect on protein expression of PR, ER alpha and ER beta. Our results showed no impact of MGD-associated sex hormones to cellular morphology and lipid accumulation in HMGEC. Cell proliferation was slightly induced through application of sex hormones and supplementation of calcium. However, both sex hormones and calcium altered gene expression of MGD-associated markers. Especially keratinization genes hornerin (HRNR) and cornulin (COR) were induced after application of sex hormones and calcium in serum-free cultivated HMGEC. This may promote keratinization processes that are associated with MGD. Further investigations are necessary to analyze the (hyper)keratinization processes that occur during MGD and using HMGEC as an in vitro model. (C) 2016 Elsevier Ltd. All rights reserved.