中华皮肤科杂志 ›› 2021, Vol. 54 ›› Issue (7): 620-624.doi: 10.35541/cjd.20201197

• 研究报道 • 上一篇    下一篇

局部外用一氧化氮供体对屏障功能破坏的小鼠表皮增生的影响

孙梦珂1    文思2    张书常1    郭盼1    王晓华2    胡立志1，3   

  1. 1天津医科大学基础医学院病原生物学系  300052；2南方医科大学皮肤病医院制剂中心，广州  510091；3天津医科大学免疫微环境与疾病省部共建教育部重点实验室  300052
  • 收稿日期:2020-12-15 修回日期:2021-05-07 发布日期:2021-07-02
  • 通讯作者: 胡立志；王晓华 E-mail:lizhihu@tmu.edu.cn; wxh_21773@163.com
  • 基金资助:
    国家自然科学基金（81573075、81972962）

Effect of topical nitric oxide donors on epidermal hyperplasia in mice with impaired barrier function

Sun Mengke1, Wen Si2, Zhang Shuchang1, Guo Pan1, Wang Xiaohua2, Hu Lizhi1,3#br#   

  1. 1Department of Pathogen Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300052, China; 2Pharmaceutical Preparation Center, Dermatology Hospital of Southern Medical University, Guangzhou 510091, China; 3Department of Immunology, Key Laboratory of Immune Microenvironment and Disease （Ministry of Education）, Tianjin Medical University, Tianjin 300052, China
  • Received:2020-12-15 Revised:2021-05-07 Published:2021-07-02
  • Contact: Hu Lizhi; Wang Xiaohua E-mail:lizhihu@tmu.edu.cn; wxh_21773@163.com
  • Supported by:
    National Natural Science Foundation of China（81573075、81972962）

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摘要： 【摘要】 目的 探讨一氧化氮（NO）对屏障功能破坏的小鼠表皮增生的影响。方法 将15只SKH1无毛小鼠按随机数字表法分为正常对照组（3只）、S-亚硝基-N-乙酰青霉胺（SNAP）处理组（4只）、屏障破坏组（4只）、屏障破坏 + SNAP处理组（4只）；将15只C57BL/6J小鼠随机分为正常对照组、屏障破坏组、屏障破坏 + 硝普钠（SNP）处理组，每组5只。正常对照组小鼠背部涂抹丙二醇-乙醇混合溶剂；SNAP处理组仅涂抹SNAP溶液；屏障破坏组采用胶带反复粘贴背部皮肤并涂抹丙二醇-乙醇混合溶剂，每天2次；屏障破坏 + SNAP或SNP处理组小鼠经胶带处理后涂抹10 mmol/L SNAP或SNP溶液。各组均连续处理4 d，第5天处死小鼠并取皮肤组织，制作石蜡切片，苏木精-伊红（HE）染色，检测表皮厚度，增殖细胞核抗原（PCNA）染色检测表皮增殖细胞。多组间比较采用双因素方差分析和单因素方差分析，两组间多重比较采用LSD-t检验。结果 SNAP处理组SKH1小鼠表皮厚度及PCNA阳性细胞数与正常对照组相比差异均无统计学意义（t值分别为0.33、1.25，P值分别为0.748、0.246）。与正常对照组相比，屏障破坏组SKH1和C57BL/6J小鼠表皮厚度均明显增加，PCNA阳性细胞数均明显增多（均P < 0.01）。屏障破坏组SKH1小鼠表皮厚度为（50.4 ± 5.4） μm，PCNA阳性细胞数为（87.3 ± 3.8）个/mm，而C57BL/6J小鼠分别为（45.9 ± 3.7） μm和（232.0 ± 19.3）个/mm，与之相比，屏障破坏 + SNAP处理组SKH1小鼠及C57BL/6J小鼠表皮均显著增厚［（127.5 ± 12.0） μm，（78.1 ± 7.6） μm，均P < 0.001］，且PCNA阳性细胞数均明显增多［（120.0 ± 5.0）个/mm，（285.0 ± 15.0）个/mm，均P < 0.01］。结论 局部NO供体处理不影响正常表皮增生，但在表皮屏障功能破坏状态下，NO供体促进表皮增生，提示皮肤状态影响局部NO供体对表皮增生的作用。

关键词: 一氧化氮, 一氧化氮供体, 表皮, 增生, 细胞增殖, 屏障功能

Abstract: 【Abstract】 Objective To evaluate the effect of nitric oxide on epidermal hyperplasia in mice with impaired barrier function. Methods Fifteen SKH1 hairless mice were divided into 4 groups by using a random number table: normal control group （3 mice）, S-nitroso-N-acetyl-DL-penicillamine （SNAP） group （4 mice）, barrier-impaired group （4 mice）, SNAP-treated barrier-impaired group （4 mice）. Fifteen C57BL/6J mice were randomly and equally divided into 3 groups: normal control group, barrier-impaired group and sodium nitroprusside （SNP）-treated barrier-impaired group. Mice in the two normal control groups were both topically treated with propylene glycol-ethanol mixtures on the back; those in the SNAP group were topically treated with SNAP solution alone; those in the two barrier-impaired groups were both treated with repeated tape peeling followed by topical application of propylene glycol-ethanol mixtures on the back twice a day; those in the SNAP- or SNP-treated barrier-impaired group were treated with repeated tape peeling followed by topical application of 10-mmol/L SNAP or SNP solution on the back twice a day. After 4 consecutive days of treatment, all the mice were sacrificed on day 5, and skin tissues were resected from the back of mice followed by preparation of paraffin sections. Hematoxylin-eosin （HE） staining was performed to measure the epidermal thickness, and proliferating cell nuclear antigen （PCNA） staining was conducted to detect proliferating cells in the epidermis. Two-way analysis of variance and one-way analysis of variance were used for comparisons among groups, and least significant difference-t test was used for multiple comparisons. Results No significant difference in the epidermal thickness or number of PCNA-positive cells was observed between the SNAP group and normal control group （t = 0.33, 1.25, P = 0.748, 0.246, respectively）. Compared with the corresponding normal control groups, the barrier-impaired groups showed significantly increased epidermal thickness and number of PCNA-positive cells （all P < 0.01）. Compared with the corresponding barrier-impaired groups, SNAP -treated barrier-impaired group and SNP-treated barrier-impaired group both showed significantly increased epidermal thickness （SKH1: 127.5 ± 12.0 μm vs. 50.4 ± 5.4 μm; C57BL/6J: 78.1 ± 7.6 μm vs. 45.9 ± 3.7 μm; both P < 0.001） and number of PCNA-positive cells （SKH1: 120.0 ± 5.0 cells/mm vs. 87.3 ± 3.8 cells/mm; C57BL/6J: 285.0 ± 15.0 cells/mm vs. 232.0 ± 19.3 cells/mm; both P < 0.01）. Conclusion Topical nitric oxide donors did not affect normal epidermis, but could aggravate epidermal hyperplasia in barrier-impaired skin, suggesting that skin condition affects the effect of topical nitric oxide donors on epidermal hyperplasia.

Key words: Nitric oxide, Nitric oxide donors, Epidermis, Hyperplasia, Cell proliferation, Barrier function