Schirra Frank, Suzuki Tomo, Dickinson Douglas P, Townsend Daniel J, Gipson Ilene K, Sullivan David A
Schepens Eye Research Institute and Harvard Medical School, Boston, MA 02114, USA.
Cornea. 2006 May;25(4):438-42. doi: 10.1097/01.ico.0000183664.80004.44.
Sex steroids exert a significant influence on the health and well-being of the ocular surface and adnexa. These hormones affect multiple aspects of the lacrimal and meibomian glands, conjunctiva, and cornea, and have been linked to the development of many ocular surface pathologies. We hypothesize that these hormone actions, as in other tissues, occur predominantly after the local synthesis of androgens and estrogens from adrenal precursors. To begin to test this hypothesis, we analyzed whether human ocular surface and adnexal tissues and cells contain the steroidogenic enzyme mRNAs necessary for the intracrine synthesis and metabolism of sex steroids.
Total RNA was isolated from human lacrimal and meibomian glands and immortalized corneal and conjunctival epithelial cells. Samples were reverse transcribed to cDNA and analyzed for the presence of enzyme mRNAs by real-time PCR. Positive and negative controls included human placental cDNA and the absence of template, respectively.
Our results show that human lacrimal and meibomian glands and corneal and conjunctival epithelial cells contain the mRNAs for steroid sulfatase, 3beta-hydroxysteroid dehydrogenase (HSD)-Delta-Delta-isomerase type 1, 17beta-HSD types 1 and 3, aromatase, and glucuronosyltransferase. In contrast, only lacrimal and meibomian tissues appeared to contain detectable mRNA for sulfotransferase.
If the corresponding mRNAs are translated, our results indicate that human ocular surface and adnexal tissues contain the enzymatic machinery necessary for the intracrine synthesis and metabolism of sex steroids.
性类固醇对眼表及附属器的健康与功能有显著影响。这些激素作用于泪腺、睑板腺、结膜和角膜的多个方面,并与多种眼表疾病的发生有关。我们推测,与其他组织一样,这些激素作用主要发生在肾上腺前体合成雄激素和雌激素之后。为了验证这一假设,我们分析了人眼表及附属器组织和细胞是否含有性类固醇内分泌合成与代谢所需的类固醇生成酶mRNA。
从人泪腺、睑板腺以及永生化角膜和结膜上皮细胞中提取总RNA。样本逆转录为cDNA,通过实时PCR分析酶mRNA的存在情况。阳性和阴性对照分别为人胎盘cDNA和无模板。
我们的结果表明,人泪腺、睑板腺以及角膜和结膜上皮细胞含有类固醇硫酸酯酶、3β-羟基类固醇脱氢酶(HSD)-Δ-Δ-异构酶1型、17β-HSD 1型和3型、芳香化酶以及葡萄糖醛酸转移酶的mRNA。相比之下,只有泪腺和睑板组织似乎含有可检测到的硫酸转移酶mRNA。
如果相应的mRNA被翻译,我们的结果表明人眼表及附属器组织含有性类固醇内分泌合成与代谢所需的酶机制。