鹅不食草醇提物通过上调 β-catenin和Gli-1促毛发生长

Ethanol extract of Centipeda minima promotes hair growth by upregulating β-catenin and Gli-1

摘要

目的 脱发严重影响患者的心理健康和生活质量。本研究旨在评价鹅不食草醇提物（ECM）对小鼠毛发生长的促进作用，并初步探讨其改善毛发生长的机制。方法 采用超声辅助方法制备ECM并利用高效液相色谱法（HPLC）检测其主要成分。将ECM 与乳膏基质混合，制得5% 和10% 浓度的ECM 乳膏。按如下分组给药：乳剂对照组、5% ECM 组、10% ECM 组、米诺地尔组，连续给药21 天，分别于第1、7、14 和21 天记录毛发生长情况。实验结束后取背部皮肤组织，进行苏木素-伊红（HE）染色、免疫组化（IHC）、免疫印记（WB）及实时荧光定量PCR （qRT-PCR）分析检测毛囊形态及β-连环蛋白（β-catenin）、神经胶质瘤相关癌基因同源物1 （Gli-1）的表达水平，探究ECM 促小鼠毛发生长效果与作用机制。结果 ECM 主要含有短叶老鹳草素A、芦丁、绿原酸、异绿原酸等成分。5% ECM 组与米诺地尔组均显著促进毛发生长，效果相当；10% ECM 组效果较弱。组织学显示，5% ECM 组毛囊数量增多、毛球膨大，接近米诺地尔组水平。分子检测表明，ECM 与米诺地尔组中β-catenin 与Gli -1的蛋白及mRNA表达均显著上调，且5% ECM组优于10% ECM组。结论 ECM可有效促进小鼠毛囊发育与毛发生长，可能与其上调β-catenin和Gli-1表达水平有关。因此，本研究为鹅不食草作为潜在的促进毛发生长药物的研发提供了理论依据。

Abstract

ObjectiveAlopecia significantly impacts patients' mental health and quality of life. This study aimed to evaluate the hair growth-promoting effect of the ethanol extract of Centipeda minima (ECM) and to preliminarily investigate its underlying mechanism. Methods ECM was prepared using ultrasonic-assisted extraction and its main constituents were examined by high performance liquid chromatography (HPLC). Subsequently, ECM was formulated into 5% and 10% creams with a cream matrix. Mice were randomly divided into four groups: vehicle control, 5% ECM, 10% ECM, and minoxidil. Treatments were applied topically for 21 consecutive days. Hair growth was monitored and recorded on days 1, 7, 14, and 21. At the end of the experiments, dorsal skin tissues were collected for hematoxylin-eosin (HE) staining, immunohistochemistry (IHC), Western blotting, and quantitative real-time PCR (qRT-PCR) analyses. These analyses were performed to examine hair follicle morphology and the expression levels of β-catenin and glioma-associated oncogene homolog 1 (Gli-1), aiming to elucidate ECM's hair growth-promoting effects and mechanisms. ResultsPhytochemical analysis revealed that the major constituents of ECM included brevilin A, rutin, chlorogenic acid, and isochlorogenic acid. Both the 5% ECM and minoxidil groups significantly promoted hair growth, exhibiting comparable efficacy, while the 10% ECM group showed a relatively weaker effect. Histological examination revealed an increased number of hair follicles and enlarged hair bulbs in the 5% ECM group, similar to the minoxidil group. Molecular analyses demonstrated significant upregulation of both protein and mRNA expression levels of β-catenin and Gli-1 in the ECM-treated and minoxidil groups, with the 5% ECM group exhibiting superior effects compared to the 10% ECM group. ConclusionECM effectively promotes hair follicle development and hair growth in mice, likely associated with its upregulation of β-catenin and Gli-1 expression. Therefore, this study provides a theoretical basis for the development of Centipeda minima as a potential hair growth-promoting drug.

关键词

鹅不食草 / 毛发生长 / 脱发 / β-连环蛋白 / 神经胶质瘤相关癌基因同源物1

Key words

Centipeda minima / Hair growth / Alopecia / β-catenin / Gli-1

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基金资助

广东省基础与应用基础研究基金项目(2021B1515140052)