环磷酰胺/紫杉醇诱导化疗脱发模型小鼠的性别间差异对比分析

doi:10.12300/j.issn.1674-5817.2026.094

实验动物与比较医学 ›› 2026, Vol. 46 ›› Issue (5): 677-688.DOI: 10.12300/j.issn.1674-5817.2026.094

环磷酰胺/紫杉醇诱导化疗脱发模型小鼠的性别间差异对比分析

钟秋明¹, 钱丽萍², 蔡媛媛², 刘德强¹, 林玮¹, 俞春英¹, 谢金东¹, 王训立¹, 吴胜², 陈碧茵³, 黄镇¹()()

^1. 福建中医药大学科技创新与转化中心, 福州 350122
^2. 福建省医学科学研究院, 福州 350001
^3. 福建中医药大学附属第三人民医院, 福州 350122

收稿日期:2026-05-19 修回日期:2026-09-07 出版日期:2026-10-25 发布日期:2026-10-16
通讯作者: 黄镇（1987—），男，博士，教授，研究方向：实验动物资源开发与利用。E-mail： zhuang@fjtcm.edu.cn。ORCID：0000-0002-8908-0355
作者简介:
钟秋明（1995—），女，硕士，助理实验师，研究方向：实验动物学。E-mail： zhongq@fjtcm.edu.cn
基金资助:
福建省第四批“雏鹰计划”青年拔尖人才项目; 福建中医药大学青年科技创新人才培育计划项目(XQC2024003)

Comparative Analysis of Sex Differences Between Cyclophosphamide- and Paclitaxel-Induced Mouse Models of Chemotherapy-Induced Alopecia

ZHONG Qiuming¹, QIAN Liping², CAI Yuanyuan², LIU Deqiang¹, LIN Wei¹, YU Chunying¹, XIE Jindong¹, WANG Xunli¹, WU Sheng², CHEN Biyin³, HUANG Zhen¹()()

^1. Innovation and Transformation Center, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China
^2. Fujian Academy of Medical Sciences, Fuzhou 350001, China
^3. The Third People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China

Received:2026-05-19 Revised:2026-09-07 Published:2026-10-25 Online:2026-10-16
Correspondence to:
HUANG Zhen (ORCID: 0000-0002-8908-0355), E-mail: zhuang@fjtcm.edu.cn

摘要/Abstract

摘要：

目的评价性别因素对环磷酰胺（cyclophosphamide，CYP）和紫杉醇（paclitaxel，PTX）诱导性脱发（chemotherapy-induced alopecia，CIA）小鼠模型表型稳定性的影响，并推测雌性小鼠表型异质性的可能机制。方法选取28只6～8周龄野生型SPF级C57BL/6J小鼠（雌雄各半），进行背部同步脱毛；脱毛后第9天，将小鼠随机分为野生型雌性CYP组、野生型雄性CYP组、雌性阴性对照组和雄性阴性对照组；其中，CYP组单次腹腔注射120 mg/kg CYP，阴性对照组腹腔注射等体积生理盐水，随后连续观察并拍照记录CYP所致CIA模型的表型变化。选取11只6周龄SPF级雌性C57BL/6J小鼠进行双侧卵巢切除（OVX组），与5只四核心基因型种鼠繁育所得XY（Sry⁻）小鼠按相同方法构建CIA模型。对OVX组、野生型雌性和雄性CYP组的小鼠于给药后第7天采集血清进行甾体激素质谱检测，以评价性腺源雌激素和性染色体背景改变对CYP诱导雌性脱发表型稳定性的影响。取20只6～8周龄SPF级C57BL/6J小鼠（雌雄各半），随机分为PTX低剂量组和PTX高剂量组，分别在同步脱毛后第9天，单次腹腔注射80 mg/kg或120 mg/kg的PTX，进行连续观察拍照，以比较雌性、雄性小鼠表型的组内一致性；再选取6只6～8周龄SPF级C57BL/6J小鼠（雌雄各半）重复PTX低剂量组的构建方法，分别于给药后当日和给药后第4、7、14、21天进行毛囊组织学检测，以观察PTX对毛囊再生的损伤。结果 CYP诱导的雄性CIA模型小鼠的表型稳定性和可重复性较高；而CYP诱导的雌性CIA模型小鼠在脱发表型上存在明显个体差异，处于动情周期的相关雌激素的组间差异可能是引发该现象的主要原因。OVX+CYP组和XY（Sry⁻）+CYP组小鼠的组间差异消失，提示卵巢来源激素及XY（Sry⁻）性染色体背景可能共同参与了雌性小鼠CYP诱导脱发表型异质性的形成。高剂量PTX组和低剂量PTX组小鼠表现出较为一致的脱发-再生过程，未见明显性别差异。结论 CYP诱导CIA模型受性别因素影响明显，卵巢切除和性染色体背景改变可提高CYP诱导雌性小鼠模型脱发表型的一致性。PTX诱导的CIA模型在雌性、雄性小鼠中均具有较好的表型一致性和性别适用性。综上，在CIA动物模型的构建、机制研究及防脱发药物评价中，应结合化疗药物类型、研究目的和表型稳定性要求合理选择实验动物性别，并将性别作为模型标准化和结果解释的重要变量。

关键词: 化疗诱导性脱发, 环磷酰胺, 紫杉醇, 性别差异, C57BL/6J小鼠, XY（Sry?）小鼠

Abstract:

Objective To evaluate the effect of sex on phenotypic stability in mouse models of chemotherapy-induced alopecia (CIA) induced by cyclophosphamide (CYP) or paclitaxel (PTX), and to explore potential mechanisms underlying phenotypic heterogeneity in female mice. Methods Twenty-eight wild-type specific pathogen-free (SPF) C57BL/6J mice (half male and half female) aged 6-8 weeks were selected for synchronous back hair removal. On the 9th day after hair removal (T0), the mice were randomly divided into a wild-type female CYP group, a wild-type male CYP group, a female negative control group and a male negative control group. Mice in the CYP groups were intraperitoneally injected with a single dose of 120 mg/kg CYP, and mice in the negative control groups were intraperitoneally injected with the same volume of normal saline. Subsequently, the phenotypic changes of the CIA model induced by CYP were continuously observed and photographed. Eleven 6-week-old SPF female C57BL/6J mice underwent bilateral ovariectomy (OVX group). Following successful surgery, these mice and five XY (Sry⁻) mice bred from Four Core Genotypes breeding stock were used to establish the chemotherapy-induced alopecia (CIA) model using the same protocol. Serum was collected from mice in the OVX group, wild-type female CYP group, and wild-type male CYP group on T7 after CYP administration for steroid hormone analysis by mass spectrometry to evaluate the effects of gonadal estrogen and sex chromosome background changes on the stability of the CYP-induced female alopecic phenotype. Finally, twenty SPF C57BL/6J mice aged 6-8 weeks (half male and half female) were randomly divided into a PTX low-dose group and a PTX high-dose group. On the 9th day after simultaneous hair removal, PTX was injected intraperitoneally at a single dose of 80 mg/kg or 120 mg/kg, respectively. Mice from the PTX low-dose group and the PTX high-dose group were continuously observed and photographed to compare the intra-group consistency of the phenotypes of female and male mice. Six SPF C57BL/6J mice (half male and half female) aged 6-8 weeks were used to repeat the construction method of the PTX low-dose group, and the hair follicle histological examination was performed at T0, T4, T7, T14 and T21, respectively, to observe the damage caused by PTX to hair follicle regeneration. Results The phenotypes of CYP-induced male CIA mice are stable and reproducible. There are obvious individual differences in the alopecia phenotype in the CYP-induced female CIA model mice, which may be mainly caused by the differences in the relevant estrogens in the estrous cycle. The difference between OVX+CYP group and XY (Sry⁻) +CYP group disappeared, suggesting that ovarian-derived hormones and XY (Sry⁻) sex chromosome background may be involved in the formation of phenotypic heterogeneity of CYP-induced alopecia in female mice. The mice in the high-dose PTX group and the low-dose PTX group showed a relatively consistent alopecia and regrowth process, and there was no significant sex difference. Conclusion CYP-induced CIA is strongly influenced by sex, and ovariectomy or alteration of the sex chromosome background can improve the consistency of CYP-induced alopecic phenotypes in female mice. PTX-induced CIA shows good phenotypic consistency in both female and male mice, indicating broad applicability across sexes. Taken together, the choice of animal sex in CIA model construction, mechanistic studies, and evaluation of anti-alopecia interventions should take into account the type of chemotherapeutic agent, the study objective, and the required degree of phenotypic stability. Sex should therefore be considered an important variable in model standardization and interpretation of experimental findings.

Key words: Chemotherapy-induced alopecia, Cyclophosphamide, Paclitaxel, Sex differences, C57BL/6J mice, XY（Sry ?） mice

中图分类号:

钟秋明,钱丽萍,蔡媛媛,等. 环磷酰胺/紫杉醇诱导化疗脱发模型小鼠的性别间差异对比分析[J]. 实验动物与比较医学, 2026, 46(5): 677-688. DOI: 10.12300/j.issn.1674-5817.2026.094.

ZHONG Qiuming,QIAN Liping,CAI Yuanyuan,et al. Comparative Analysis of Sex Differences Between Cyclophosphamide- and Paclitaxel-Induced Mouse Models of Chemotherapy-Induced Alopecia[J]. Laboratory Animal and Comparative Medicine, 2026, 46(5): 677-688. DOI: 10.12300/j.issn.1674-5817.2026.094.

图/表 7

图1 化疗诱导性脱发小鼠模型的构建流程及观察时间点

Figure 1 Construction workflow and observation time points of the chemotherapy-induced alopecia mouse model

图2 环磷酰胺诱导的化疗性脱发小鼠模型中雌性、雄性表型差异的观察结果

注：于第0天（Day 0）进行背部同步脱毛；背部同步脱毛后第9天，野生型雄性环磷酰胺（CYP）组（n=5）和野生型雌性CYP组（n=5）小鼠均单次腹腔注射120 mg/kg CYP，对照组（n=6）小鼠腹腔注射等体积生理盐水；分别于给药后第0天（T0）、第4天（T4）、第7天（T7）、第14天（T14）和第21天（T21），观察各组小鼠的背部被毛情况。红色方框表示代表性的不明显脱发表型。

Figure 2 Observations of phenotypic differences between females and males in a cyclophosphamide-induced mouse model of chemotherapy-induced alopecia

Note： On Day 0， perform simultaneous back hair removal. On day 9 after simultaneous back hair removal， the wild-type male cyclophosphamide （CYP） group （n=5） and the wild-type female CYP group （n=5） mice were each given a single intraperitoneal injection of 120 mg/kg CYP， while the control group （n=6） mice were given the same volume of normal saline intraperitoneally. At time points of Day 0 （T0）， Day 4 （T4）， Day 7 （T7）， Day 14 （T14）， and Day 21 （T21） after administration， observe the back hair condition of each group of mice. Red boxes indicate representative examples of inconspicuous alopecia.

图3 野生型雌性环磷酰胺组和野生型雄性环磷酰胺组小鼠的脱发评分结果

注：背部同步脱毛后第9天（即T0），野生型雄性CYP组和野生型雌性环磷酰胺（CYP）组小鼠单次腹腔注射120 mg/kg CYP。T7、T14分别为CYP腹腔注射后第7天和第14天。雄性和雌性小鼠各5只，T7和T14为同一批动物重复观察；散点代表单只小鼠评分值。

Figure 3 Alopecia score results of wild-type female cyclophosphamide-treated and wild-type male cyclophosphamide-treated group mice

Note： On day 9 after synchronized dorsal depilation， mice in the wild-type male and female cyclophosphamide （CYP） groups received a single intraperitoneal injection of CYP （120 mg/kg）. T7 and T14 were days 7 and 14 after CYP intraperitoneal injection， respectively. Five mice of each sex were included. The same animals were evaluated at both T7 and T14， and each dot represents the score of an individual mouse.

图4 野生型雌性环磷酰胺组、野生型雄性环磷酰胺组和卵巢切除鼠环磷酰胺干预组小鼠血清中的甾体激素水平

注：腹腔注射120 mg/kg环磷酰胺（CYP）后第7天（T7），野生型雌性CYP组、野生型雄性CYP组和卵巢切除鼠CYP干预组（OVX+CYP）小鼠血清中孕酮（A）、17β-雌二醇（B）和睾酮（C）水平。先通过双侧卵巢切除术构建OVX模型，术后恢复1周；再对OVX小鼠、野生型小鼠背部同步脱毛；脱毛后第9天（T0），野生型雄性CYP组（n=6）、野生型雌性CYP组（n=6）小鼠和OVX+CYP组（n=6）小鼠单次腹腔注射120 mg/kg CYP。散点代表单只小鼠的检测结果，^ns P>0.05，^* P<0.05；^*** P<0.001。

Figure 4 Steroid hormone levels in serum of wild-type female cyclophosphamide group， wild-type male cyclophosphamide group， and ovariectomized + cyclophosphamide group mice

Note： Serum levels of progesterone （A）， 17β-estradiol （B）， and testosterone （C） in the wild-type female cyclophosphamide （CYP）， wild-type male CYP， and ovariectomized CYP-treated （OVX+CYP） groups on day 7 （T7） after intraperitoneal administration of cyclophosphamide （120 mg/kg）. The OVX model was established by bilateral ovariectomy， followed by a 1-week recovery period. OVX and wild-type mice then underwent synchronized dorsal depilation. On day 9 after depilation （T0）， mice in the wild-type male CYP （n=6）， wild-type female CYP （n=6）， and OVX+CYP （n=6） groups received a single intraperitoneal injection of CYP （120 mg/kg）. Each dot represents an individual mouse； ^ns P>0.05， ^* P<0.05， ^*** P<0.001.

图5 卵巢切除和睾丸决定基因Sry -缺失可提高环磷酰胺诱导脱发小鼠模型的表型一致性

注：卵巢切除鼠环磷酰胺干预（OVX+CYP）组和XY（Sry⁻）+CYP组小鼠在不同观察时间点的背部被毛变化。OVX+CYP组小鼠先行双侧卵巢切除术，术后恢复1周；随后OVX+CYP组与XY（Sry⁻）+ CYP组小鼠进行背部同步脱毛；同步脱毛后第9天，OVX+CYP组（n=5）和XY（Sry⁻）+CYP组（n=5）小鼠均单次腹腔注射120 mg/kg CYP。Day 0和Day 7分别为同步脱毛后第0、7天，T0、T4、T7、T14和T21分别为CYP给药后第0、4、7、14和21天。

Figure 5 Ovariectomy and deletion of the testis determining gene Sry - can improve the phenotypic consistency of the cyclophosphamide-induced mouse model of chemotherapy-induced alopecia

Note： Changes in dorsal hair coverage in the OVX+cyclophosphamide （CYP） and XY （Sry⁻）+CYP groups at different observation time points. Mice in the OVX+CYP group underwent bilateral ovariectomy followed by a one-week recovery period. Mice in both the OVX+CYP and XY （Sry⁻）+CYP groups then underwent synchronized dorsal depilation. On day 9 after depilation （T0）， mice in the OVX+CYP group （n=5） and XY （Sry⁻）+CYP group （n=5） received a single intraperitoneal injection of CYP （120 mg/kg）. Day 0 and Day 7 were the 0 and 7 days after simultaneous hair removal. T0， T4， T7， T14， and T21 correspond to days 0， 4， 7， 14， and 21 after CYP administration， respectively.

图6 紫杉醇诱导的化疗性脱发模型中雌性、雄性小鼠的表型差异观察结果

注：PTX低剂量组和PTX高剂量组小鼠在腹腔注射紫杉醇（PTX）后不同时间点的背部被毛变化。小鼠背部同步脱毛后第9天（即T0），PTX低剂量组（80 mg/kg，n=10）和PTX高剂量组（120 mg/kg，n=10）分别单次腹腔注射相应剂量PTX。Day 0和Day 7分别为同步脱毛后第0、7天；T0、T4、T7、T14和T21分别为PTX给药后第0、4、7、14和21天。本图所示低、高剂量表型观察亚组均为雌雄各5只。

Figure 6 Phenotypic differences between female and male mice in a paclitaxel-induced chemotherapy alopecia mouse model

Note： Changes in dorsal hair coverage in the low-dose and high-dose paclitaxel （PTX） groups at different time points after intraperitoneal PTX administration. On day 9 （T0） after synchronized dorsal depilation， mice in the low-dose PTX group （80 mg/kg， n=10） and high-dose PTX group （120 mg/kg， n=10） received a single intraperitoneal injection of PTX at the respective doses. Day 0 and Day 7 were the 0 and 7 days after simultaneous hair removal， respectively. T0， T4， T7， T14 and T21 were days 0， 4， 7， 14 and 21 after PTX administration， respectively. For the phenotypic assessments shown in this figure， both the low- and high-dose PTX groups included five male and five female mice.

图7 低剂量紫杉醇诱导的化疗性脱发小鼠模型毛囊组织的HE染色结果

注：6只SPF级C57BL/6J小鼠（雌雄各半），背部同步脱毛后第9天，分别单次腹腔注射80 mg/kg PTX，并在给药后第0天（T0）、第4天（T4）、第7天（T7）、第14天（T14）和第21天（T21）进行连续观察。标尺=100 μm。

Figure 7 HE staining of hair follicle tissues in the low-dose paclitaxel-induced chemotherapy alopecia mouse model

Note： Six SPF C57BL/6J mice （half male and half female） were given a single intraperitoneal injection of 80 mg/kg PTX on the 9th day after synchronous back hair removal， and continuously observed on days 0 （T0）， 4 （T4）， 7 （T7）， 14 （T14） and 21 （T21） after administration. Scale bar=100 μm.

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环磷酰胺/紫杉醇诱导化疗脱发模型小鼠的性别间差异对比分析

Comparative Analysis of Sex Differences Between Cyclophosphamide- and Paclitaxel-Induced Mouse Models of Chemotherapy-Induced Alopecia

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