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Let-7b 通过下调外胚层发育蛋白 A 来调节羊驼的毛发生长。

Let-7b regulates alpaca hair growth by downregulating ectodysplasin A.

机构信息

Alpaca Bioengineering Laboratory, College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi 030801, P.R. China.

出版信息

Mol Med Rep. 2018 Mar;17(3):4688-4694. doi: 10.3892/mmr.2018.8442. Epub 2018 Jan 17.

Abstract

Hypohidrotic ectodermal dysplasia (HED), also known as anhidrotic ectodermal dysplasia, is characterized by the clinical manifestations of less sweat or no sweat, sparse or no hair, tooth agenesis and/or abnormal tooth morphology. The characteristics of alpaca ear hair differ from the back hair. The ectodysplasin A (EDA) signaling pathway has a regulatory effect on skin development and hair growth. The aim of the present study was to study the effects of EDA on alpaca hair growth by examining the mRNA and protein expression levels of EDA in alpaca ear and back skin by reverse transcription‑quantitative polymerase chain reaction and western blot analysis, respectively. Results indicated that EDA expression was higher in the ear skin compared with the back skin. The expression levels of let‑7b in the skin of healthy alpacas varies; the difference between let‑7b expression levels of the ear and back have been reported to be >2‑fold, suggesting a role for let‑7b in the development of adult alpaca skin and hair follicles. A dual‑luciferase reporter vector was constructed to verify the targeting relationship between microRNA let‑7b and EDA, and the results revealed that EDA was a target gene of let‑7b. Alpaca skin fibroblasts were transfected with a let‑7b eukaryotic expression vector to investigate the regulatory relationship between let‑7b and EDA. The expression of EDA was decreased in the transfected group; immunocytochemical results demonstrated that the EDA protein was abundantly expressed in the fibroblast cytoplasm. EDA protein expression was weaker in the transfected cells than in the untransfected cells. These results suggested that EDA may serve a role in alpaca hair growth and is probably a target gene of let‑7b; let‑7b downregulated EDA mRNA and protein expressions, which suggested that let‑7b may regulate alpaca hair growth. These conclusions suggested that let‑7b may be associated with HED.

摘要

少汗型外胚层发育不全(HED),又称无汗性外胚层发育不全,其临床表现为少汗或无汗、毛发稀疏或无毛发、牙齿缺失和/或牙齿形态异常。羊驼耳毛的特征不同于背部毛发。外胚层蛋白 A(EDA)信号通路对皮肤发育和毛发生长具有调节作用。本研究旨在通过反转录-定量聚合酶链反应和 Western blot 分析分别检测羊驼耳和背部皮肤中 EDA 的 mRNA 和蛋白表达水平,研究 EDA 对羊驼毛发生长的影响。结果表明,EDA 在耳皮肤中的表达高于背部皮肤。健康羊驼皮肤中 let-7b 的表达水平存在差异;耳和背部的 let-7b 表达水平差异>2 倍,提示 let-7b 在成年羊驼皮肤和毛囊发育中起作用。构建了双荧光素酶报告载体,以验证 microRNA let-7b 与 EDA 的靶向关系,结果显示 EDA 是 let-7b 的靶基因。转染羊驼皮肤成纤维细胞 let-7b 真核表达载体,研究 let-7b 与 EDA 之间的调控关系。转染组 EDA 表达下降;免疫细胞化学结果表明 EDA 蛋白在成纤维细胞细胞质中大量表达。转染细胞中的 EDA 蛋白表达弱于未转染细胞。这些结果表明 EDA 可能在羊驼毛发生长中起作用,可能是 let-7b 的靶基因;let-7b 下调 EDA mRNA 和蛋白表达,提示 let-7b 可能调节羊驼毛发生长。这些结论提示 let-7b 可能与 HED 有关。

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