Effects of Pine Massoniana Needle Extract on Androgenetic Alopecia Model Rats via Nrf2-ARE Mediated Signaling Pathway
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摘要:
目的 观察马尾松针提取物对双氢睾酮(Dihydrotestosterone, DHT)诱导的雄激素性脱发(Androgenetic alopecia, AGA)模型小鼠的干预效应, 并通过核因子E2相关因子2(Nrf2)-抗氧化反应元件(ARE)信号通路探讨马尾松针提取物促毛发生长的作用机制。 方法 采用DHT皮下注射建立AGA小鼠模型, 48只雄性C57BL/6小鼠随机分为空白组, DHT组, 原花青素组, 马尾松针提取物(松针)低、中、高剂量组, 每组8只。空白组不予任何处理, DHT组仅皮下注射DHT 30 mg·d-1, 原花青素组皮下注射DHT并灌服原花青素B2 5 mg·kg-1, 松针低、中、高剂量组皮下注射DHT, 同时分别灌服马尾松针提取物4、8、12 mg·kg-1。每组均给药4周, 每周拍照并记录毛发生长情况。d28后取其背部圆形皮片, 刮取毛发称质量, 行HE染色。计算生长期、休止期毛囊数量并计算二者比值。取部分皮肤组织, 检测ROS和MDA含量。qPCR和Western blot法检测Nrf2、Keap1、NQO1、HO-1、TGF-β1 mRNA与蛋白的表达水平。 结果 马尾松针提取物呈剂量依赖性地促进AGA小鼠的毛发生长, 松针高剂量组与原花青素组毛发质量以及生长期/休止期毛囊比值均无显著性差异。马尾松针a提取物能够降低组织内ROS和MDA含量, 呈剂量依赖性地促进Nrf2、NQO1、HO-1 mRNA与蛋白的表达, 抑制Keap1、TGF-β1 mRNA与蛋白的表达。 结论 马尾松针提取物可能通过激活Nrf2-ARE信号通路, 改善氧化应激水平, 从而促进AGA小鼠毛发的生长。 -
关键词:
- 马尾松针提取物 /
- 雄激素性脱发 /
- 氧化应激 /
- Nrf2-ARE通路
Abstract:OBJECTIVE To investigate the hair growth-promoting activity and the possible mechanism of pine massoniana needle extract (PMNE) on mice by using the dihydrotestosterone (DHT) induced androgenetic alopecia (AGA) model. METHODS 48 C57BL/6 mice were randomly divided into 6 groups: control group without any treatment, DHT group (30 mg ·d-1 DHT only), procyanidin group (DHT+procyanidin B2 5 mg ·kg-1), PMNE low dose group (DHT+PMNE 4 mg ·kg-1), PMNE medium dose group (DHT+PMNE 8 mg ·kg-1) and PMNE high dose group (DHT+PMNE 12 mg ·kg-1). PMNE and procyanidin B2 were given orally every day. The hair growth of mice was observed and evaluated at d7, d14, d21 and d28 after administration. The circle skins of all mice were harvested at the same depilated aera after 28 days. The hair from these skins was weighed. Some skin tissue section was obtained for HE staining, and the number of hair follicles were calculated. Some skin tissues were obtained to detect the contents of ROS and MDA and the mRNA and protein levels of Keap1, Nrf2, NQO1, HO-1 and TGF-β1 by qPCR and Western blot. RESULTS PMNE promoted the hair growth of AGA mice and showed a dose-dependent effect. PMNE of 12 mg ·kg-1 could alleviate the DHT-induced AGA in mice and achieve a similar effect to procyanidin B2. PMNE reduced the contents of ROS and MDA, promoted the mRNA and protein expressions of Nrf2, NQO1, HO-1 and reduced the mRNA and protein expressions of Keap1 and TGF-β1. CONCLUSIONS PMNE can promote the hair growth of AGA mice by regulating the Nrf2-ARE signaling pathway. -
表 1 引物序列
Table 1. The sequence of primer
基因 正向引物(5'→3') 反向引物(5'→3') 长度/bp Nrf2 CTGAACTCCTGGACGGGACTA CGGTGGGTCTCCGTAAATGG 20 Keap1 TCGAAGGCATCCACCCTAAG CTCGAACCACGCTGTCAATCT 21 NQO1 TACGACAACGGTCCTTTCCAG ACAGAAACGCAGGATGCCACT 21 HO-1 AAGCCGAGAATGCTGAGTTCA GCCGTGTAGATATGGTACAAGGA 23 TGF-β1 CTCCCGTGGCTTCTAGTGC GCCTTAGTTTGGACAGGATCTG 22 β-actin GGCTGTATTCCCCTCCATCG CCAGTTGGTAACAATGCCATGT 22 表 2 各组小鼠背部皮肤毛囊数量比较(x±s, n=8)
Table 2. Comparison of the number of hair follicles in each group (x±s, n=8)
组别 生长期 休止期 生长期/休止期 空白组 74.13±31.00 9.63±2.39 7.25±2.81 DHT组 12.38±3.38** 7.88±4.22 2.13±1.25* 原花青素组 64.25±19.91## 7.50±2.14 9.88±5.46# 松针低剂量组 27.63±15.21 7.00±4.99 6.88±9.05 松针中剂量组 27.63±13.10 5.38±3.16* 6.63±3.46 松针高剂量组 62.25±28.40## 5.88±3.44* 14.13±11.03## 注: 与空白组比较, *P < 0.05, **P < 0.01;与DHT组比较, #P < 0.05, ##P < 0.01。 -
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