CN114848625B - Ferrostatin-1在制备促进毛发生长的试剂中的应用 - Google Patents
Ferrostatin-1在制备促进毛发生长的试剂中的应用 Download PDFInfo
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Abstract
本发明提供了Ferrostatin‑1在制备促进毛发生长的试剂中的应用，涉及生发技术领域。本发明对脱毛的小鼠给予Fer‑1，相比于对照组，小鼠毛发生长较快，毛发更为浓密，且毛囊数也增多，证实所述Ferrostatin‑1具有促进毛发生长的作用。
Description
技术领域
本发明属于生发技术领域，具体涉及Ferrostatin-1在制备促进毛发生长的试剂中的应用。
背景技术
Ferrostain-1(Fer-1，MCE)是一种人工合成的抗氧化剂，具有抑制脂质过氧化的作用，是一种强大的铁死亡抑制剂。文献报道Fer-1是一种脂质活性氧清除剂，能抑制铁死亡诱导剂(Erastin)诱导的胞浆和脂质活性氧的积累。目前尚无文献报道其促毛发生长的作用。
发明内容
有鉴于此，本发明的目的在于提供Ferrostatin-1在制备促进毛发生长的试剂中的应用，所述Ferrostatin-1具有良好的刺激毛囊生长的作用，可用于刺激毛发生长。
为了实现上述发明目的，本发明提供以下技术方案：
本发明提供了Ferrostatin-1在制备促进毛发生长的试剂中的应用。
优选的，所述Ferrostatin-1的工作浓度低于5μM。
优选的，所述试剂的类型包括外用试剂。
本发明还提供了一种生发剂，有效成分包括Ferrostatin-1，还包括辅料。
有益效果：本发明提供了Ferrostatin-1在制备促进毛发生长的试剂中的应用，实施例中用Fer-1涂抹小鼠，探讨Fer-1是否能促进毛发生长，结果显示，给予Fer-1的小鼠毛发生长较快且毛发更为浓密，其中0.5μM Fer-1组较5μM Fer-1组更快进入生长期；0.5μMFer-1组和5μM Fer-1组较对照组总毛囊数增加，且0.5μM Fer-1组毛囊数多于5μM Fer-1组，说明低浓度的Fer-1有较好的促进毛发生长的作用。
附图说明
图1为小鼠毛发生长情况；
图2为小鼠毛发评分标准图；
图3为小鼠毛发评分图；
图4为小鼠皮肤H&E染色结果。
具体实施方式
本发明提供了Ferrostatin-1在制备促进毛发生长的试剂中的应用。
本发明所述Fer-1优选在低浓度下使用，更优选在低于5μM的浓度时使用；且所述试剂优选为外用试剂。
本发明实施例中，以脱毛的小鼠为模型进行研究，对模型小鼠涂抹所述Fer-1，与对照组相比，给予Fer-1的小鼠毛发生长较快且毛发更为浓密，其中0.5μM Fer-1组第7天开始进入生长期，5μM Fer-1组则是第8天进入生长期；实验后测定小鼠皮肤毛囊数量，2％米诺地尔组、0.5μM Fer-1组和5μM Fer-1组较对照组总毛囊数增加，且0.5μM Fer-1组毛囊数多于5μM Fer-1组，说明低浓度的Fer-1有较好的促进毛发生长的作用。
本发明还提供了一种生发剂，有效成分包括Ferrostatin-1，还包括辅料。
本发明所述生发剂优选为外用试剂，所用辅料为本领域的常规辅料，在此不做特殊限定。
下面结合实施例对本发明提供的Ferrostatin-1在制备促进毛发生长的试剂中的应用进行详细的说明，但是不能把它们理解为对本发明保护范围的限定。
实施例1
一、实验过程
1.实验材料和仪器
1.1实验动物
C57BL/6雌性小鼠，体重18～22g，购自于北京华阜康生物科技股份有限公司(许可证号：SCXK(京)2019-0008)，动物饲养于遵义医科大学基础药理教育部重点实验室SPF动物房，各组实验动物自由进食、饮水。
1.2主要仪器设备
组织包埋机(Leica)；切片机(Leica)；自动染片机(Leica)；Olympus光学显微镜(日本Olympus公司)；Milli QA纯水处理器(Millipore公司)等。
1.3主要实验试剂
Ferrostatin-1(Fer-1，MCE)；2％米诺地尔(蔓迪)
2.实验方法
2.1实验分组
40只C57BL/6小鼠，雌性，体重18～22g，随机分为4组，每组10只，分别设为对照组(Control)、2％米诺地尔组、0.5μM Fer-1组和5μM Fer-1组。SPF级动物房适应性饲养一周。
2.2给药方法
使用动物剃毛器将小鼠背部皮肤的毛发剃除，用脱毛膏涂抹于剃毛区域，等待约3min后将脱毛膏洗净擦干，诱导小鼠毛发进入休止期(此时小鼠皮肤为粉红色)，拍照记录初始状态。采用背部涂抹给药方法，每组每只给药200μL，每日给药一次，直至各组小鼠毛发生长呈现显著性差异时停止给药。
2.3毛发生长情况观察
给药后逐日观察各组动物脱毛区域皮肤及毛发生长情况，并记录每只小鼠脱毛区皮肤颜色由粉红变成黑色的时间及开始长毛的时间，评价药物对小鼠毛发生长的影响。从皮肤颜色变化的时间起对每组拍照并记录毛发生长情况评分结果，评分标准如下：
0分：未生长，脱毛区皮肤呈粉色；1分：脱毛区局部皮肤呈灰色(小于30％的区域)；2分：脱毛区皮肤全呈灰色；3分：脱毛区皮肤局部呈黑色(60％～80％的区域)；4分：脱毛区皮肤全呈黑色并有少许毛发生长(40％～60％的生长)；5分：脱毛区皮肤呈黑色并有部分毛发生长(60％～80％的生长)。
2.4取材和标本制备
各组小鼠于给药结束后第二天腹腔注射2％戊巴比妥钠(0.05mL/10g)麻醉，刮去表面细毛后，在给药部位平行于脊椎处取材，将皮肤修剪成1cm宽的长条状，置于4％多聚甲醛固定24～36h。
2.5H&E染色
(1)样本用4％多聚甲醛固定24h以上后，置于流水冲洗30min。
(2)脱水：将组织浸泡在体积浓度为75％乙醇、85％乙醇、95％乙醇I、95％乙醇II、无水乙醇I和无水乙醇II中，各10min。
(3)透明：将脱水后的组织分别浸泡在二甲苯I和二甲苯II中各10min。
(4)透蜡：将透明处理过的组织浸泡于融化的石蜡中3h。
(5)包埋。
(6)切片：使用切片机将组织切成5μm厚度的组织片，使其紧贴载玻片上。
(7)烤片：将切片放置于65℃烘箱中烤片1h。
(8)脱蜡复水：二甲苯I、二甲苯II各10min，无水乙醇、95％乙醇、85％乙醇、75％乙醇、蒸馏水各2min。
(9)苏木素染色：苏木精染料10min，流水冲洗3s，镜检，若苏木素着色过深可用l％盐酸酒精进行分脱色3s，脱去细胞质中多余的苏木素染色液，然后自来水3min。
(10)伊红染液染色：染色2min、95％乙醇脱水5min、二甲苯10min。
(11)封片：中性树脂封片。
(12)镜检：细胞核蓝色、细胞质红色。
2.6数据的统计分析
采用SPSS 26软件对数据进行统计分析处理，所有数据以均数±标准误(Mean±SEM)表示，采用单因素方差分析(ANOVA)比较均数的组别差异，方差齐采用LSD法进行组间两两比较，不齐则采用Dunnett’s T3检验，P<0.05有统计学意义。
实验结果显示：图1是小鼠毛发生长情况，图2是小鼠毛发评分标准图，图3是小鼠毛发评分图，如图1和图2所示，与对照组相比，给予Fer-1的小鼠毛发生长较快且毛发更为浓密，其中0.5μM Fer-1组第7天开始进入生长期，5μM Fer-1组则是第8天进入生长期。图4为小鼠皮肤纵切面H&E染色结果，皮肤横切面用于测定毛囊数量，2％米诺地尔组、0.5μMFer-1组和5μM Fer-1组较对照组总毛囊数增加，且0.5μM Fer-1组毛囊数多于5μM Fer-1组，说明低浓度的Fer-1有较好的促进毛发生长的作用。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (3)
1.Ferrostatin-1在制备促进毛发生长的试剂中的应用。
2.根据权利要求1所述应用，其特征在于，所述Ferrostatin-1的工作浓度低于5μM。
3.根据权利要求1或2所述应用，其特征在于，所述试剂的类型包括外用试剂。
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| CN113181145A (zh) * | 2021-05-25 | 2021-07-30 | 浙江大学医学院附属邵逸夫医院 | 姜酮在制备治疗脱发的药物或制剂中的应用 |
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