超活化富血小板裂解液的制备及其对雄激素性脱发小鼠模型毛囊再生作用

刘春香; 王博昊; 张怡

doi:10.3760/cma.j.issn.1673-4211.2020.01.004

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国际生物制品学杂志

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ENGLISH ABSTRACT

超活化富血小板裂解液的制备及其对雄激素性脱发小鼠模型毛囊再生作用

作者及单位信息

出版日期： 2020 -02 -10 ·

DOI： 10.3760/cma.j.issn.1673-4211.2020.01.004

Preparation of super-activated platelet lysate and its effect on hair follicle regeneration in a mouse model of androgenetic alopecia

Authors Info & Affiliations

Liu Chunxiang

National and Local Joint Stem Cell Research & Engineering Center for Aging Diseases, Heilongjiang Tianqing Stem Cell Co., Ltd., Harbin 150028, China

Wang Bohao

Zhang Yi

Published： 2020 -02 -10 ·

DOI： 10.3760/cma.j.issn.1673-4211.2020.01.004

709

154

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摘要

目的制备超活化富血小板裂解液(super-activated platelet lysate，sPL)，研究其对雄激素性脱发(androgenetic alopecia，AGA)小鼠模型的毛囊再生的影响，为sPL治疗AGA的临床研究提供基础。

方法使用超滤离心从全血中分离富血小板血浆(platelet-rich plasma，PRP)，用冻融方式将血小板裂解制备血小板裂解液(platelet lysate，PL)。用CaCl ₂结合肝素激活PRP中的血小板，冻融裂解血小板的同时用温度浮动控制方法去除纤维蛋白原，获得sPL。通过ELISA分析PRP、PL及sPL中毛囊再生相关因子含量。用丙酸睾酮注射建立C57bl/6小鼠AGA模型后，用sPL进行治疗，取小鼠背部皮肤，苏木素伊红染色判断sPL对毛囊再生的作用。

结果经超滤离心及激活获得的sPL中，数种毛囊再生相关因子较PRP及PL均显著增加。AGA小鼠经sPL治疗后毛发再生明显，同时毛囊密度恢复至对照组水平( t＝1.149， P＝0.269 9)。

结论相对于PRP与PL，sPL制备方法可获得更高的生长因子含量，且sPL对AGA小鼠具有显著的毛囊再生作用。

关键词：秃发;毛囊再生;超活化富血小板裂解液

ABSTRACT

ObjectiveTo prepare super-activated platelet lysate (sPL) and study its effect on hair follicle regeneration in a mouse model of androgenetic alopecia (AGA), providing basis for AGA treatment research.

MethodsPlatelet-rich plasma (PRP) was isolated from whole blood by ultrafiltration centrifugation. Concentrated platelets were frozen and thawed to obtain platelet lysate (PL). To prepare sPL, first platelet activation was performed by addition of CaCl ₂ in combination with heparin, then the sample was completely lysed by freeze-thaw method and fibrinogen was depleted by temperature controlled clotting procedure. Hair follicle regeneration-related growth factors in PRP, PL and sPL were measured by ELISA. C57bl/6 AGA mouse model was established by administration of testosterone propionate. Hair follicle regeneration after treatment of sPL was revealed by hemotoxilin and eosin staining.

ResultsThe amount of several hair regeneration-related growth factors in sPL obtained with ultrafiltration centrifugation and platelet activation was significantly higher than that in PRP and PL. AGA mice evidently regrowed hair after sPL treatment, and hair follicle density increased to the level of control mice ( t=1.149, P=0.269 9).

ConclusionsComparing with PRP and PL, sPL preparation can result in higher growth factor concentrations, and sPL has significant hair follicle regeneration effect on AGA mouse model.

KEYWORDS： Alopecia;Hair follicle regeneration;Super-activated platelet lysate

Corresponding author: Zhang Yi, Email: mocdef.3ab61gnahz_iy_oen

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引用本文

刘春香,王博昊,张怡. 超活化富血小板裂解液的制备及其对雄激素性脱发小鼠模型毛囊再生作用[J]. 国际生物制品学杂志,2020,43(01)：16-19.

DOI:10.3760/cma.j.issn.1673-4211.2020.01.004

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雄激素性脱发(androgenetic alopecia，AGA)是临床最常见的一种雄激素依赖的多基因遗传病 ^{[
1
]}，症状为进行性毛发稀疏与毛囊萎缩。AGA发病特征为毛发的生长期显著缩短，使毛囊转变成类毫毛毛囊，增加了休止期头发的脱落，毛发提前终止生长期而进入退化期 ^{[
2
]}。

目前，AGA治疗仍以中药调理及药物治疗(米诺地尔和非那雄胺等)为主，这些治疗方法可调节雄激素过量表达或阻断激素受体以改善脱发，但均未能对已产生脱发的微环境进行调理 ^{[
3
]}。富血小板血浆(platelet-rich plasma，PRP)含有大量生长因子，能有效刺激毛囊细胞生长、改善头皮微环境及头皮血供 ^{[
4
]}。本研究通过将PRP有效激活与裂解，释放生长因子制备超活化富血小板裂解液(super-activated platelet lysate，sPL)后，治疗AGA动物模型，为AGA临床研究提供基础。

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摘要
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备注信息

通信作者：张怡，Email: mocdef.3ab61gnahz_iy_oen

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