大麻素受体2抑制对口腔黏膜天疱疮棘层松解影响的研究

doi:10.7518/hxkq.2025.2025182

华西口腔医学杂志 ›› 2025, Vol. 43 ›› Issue (6): 829-836.doi: 10.7518/hxkq.2025.2025182

• 基础研究 • 上一篇

大麻素受体2抑制对口腔黏膜天疱疮棘层松解影响的研究

刘慧娟¹, 宋鹏², 侯亚丽², 霍晓³, 秘利进⁴, 刘春艳⁵()

^1.河北医科大学口腔医（学）院重点实验室，河北省口腔医学重点实验室，河北省口腔健康技术创新中心，石家庄 050017
^2.河北医科大学口腔医（学）院病理科，河北省口腔医学重点实验室，河北省口腔健康技术创新中心，石家庄 050017
^3.河北医科大学口腔医（学）院黏膜科，河北省口腔医学重点实验室，河北省口腔健康技术创新中心，石家庄 050017
^4.河北医科大学口腔医（学）院检验科，河北省口腔医学重点实验室，河北省口腔健康技术创新中心，石家庄 050017
^5.河北医科大学口腔医（学）院正畸科，河北省口腔医学重点实验室，河北省口腔健康技术创新中心，石家庄 050017

收稿日期:2025-04-24 修回日期:2025-07-14 出版日期:2025-12-01 发布日期:2025-11-27
通讯作者: 刘春艳 E-mail:chunyanliu@hebmu.edu.cn
作者简介:刘慧娟，副主任医师，博士，E-mail：huijuan_liu_001@hebmu.edu.cn
基金资助:
河北省医学科学研究课题(20241093)

Cannabinoid receptor 2 inhibition on acantholysis in oral mucosal pemphigus

Liu Huijuan¹, Song Peng², Hou Yali², Huo Xiao³, Mi Lijin⁴, Liu Chunyan⁵()

^1.Key Laboratory of Stomatology, School and Hospital of Stomatology, Hebei Medical University & Hebei Key Laboratory of Stomatology, Hebei Technology Innovation Center of Oral Health, Shijiazhuang 050017, China
^2.Dept. of Pathology, School and Hospital of Stomatology, Hebei Medical University & Hebei Key Laboratory of Stomato-logy, Hebei Technology Innovation Center of Oral Health, Shijiazhuang 050017, China
^3.Dept. of Oral Medicine, School and Hospital of Stomatology, Hebei Medical University & Hebei Key Laboratory of Stomatology, Hebei Technology Innovation Center of Oral Health, Shijiazhuang 050017, China
^4.Dept. of Clinical Laboratory, School and Hospital of Stomatology, Hebei Medical University & Hebei Key Laboratory of Stomatology, Hebei Technology Innovation Center of Oral Health, Shijiazhuang 050017, China
^5.Dept. of Orthodontics, School and Hospital of Stomatology, Hebei Medical University & Hebei Key Laboratory of Stomatology, Hebei Technology Innovation Center of Oral Health, Shijiazhuang 050017, China

Received:2025-04-24 Revised:2025-07-14 Online:2025-12-01 Published:2025-11-27
Contact: Liu Chunyan E-mail:chunyanliu@hebmu.edu.cn
Supported by:
Medical Science Research Project of Hebei(20241093)

摘要/Abstract

摘要：

目的利用大麻素受体2（CB2）抑制剂和天疱疮血清共培养HaCaT细胞，探讨CB2抑制对HaCaT细胞桥粒黏蛋白3（DSG3）的影响。方法采用免疫组织化学染色观察天疱疮患者和正常受试者黏膜中CB受体表达，酶联免疫吸附实验（ELISA）检测天疱疮患者和正常受试者血清中CB2含量，对天疱疮患者血清CB2浓度与DSG表达进行相关性分析。CCK-8检测不同浓度AM630对HaCaT细胞的抑制作用，计算AM630对HaCaT细胞的半数抑制浓度（IC₅₀）并作为抑制剂实验浓度。将正常受试者、天疱疮患者血清分别与完全培养液1∶1比例共培养HaCaT细胞作为阴性对照组和天疱疮组，完全培养液培养的HaCaT细胞作为对照组，天疱疮组培养液稀释AM630至IC₅₀浓度后培养的HaCaT细胞作为AM630组。实时荧光定量聚合酶链反应（PCR）、蛋白印迹（Western blot）检测4组CB2、DSG3、β-catenin基因和蛋白的表达。细胞解离实验观察AM630对HaCaT细胞黏附功能的影响。结果免疫组织化学染色显示天疱疮黏膜与正常黏膜组织中CB2表达差异有统计学意义（P<0.000 1），CB1表达差异无统计学意义。ELISA检测表明，正常受试者与天疱疮患者血清CB2的表达差异有统计学意义（P<0.001）。天疱疮患者血清CB2表达与DSG3表达呈正相关（r=0.831，P=0.003）。CCK-8检测表明，AM630对HaCaT细胞的半数抑制浓度IC₅₀为0.55 μmol/L。实时荧光定量PCR、Western blot结果显示，天疱疮组CB2、DSG3表达升高，β-catenin表达降低，与AM630组差异有统计学意义（P<0.05）。结论口腔黏膜天疱疮中CB2高表达，AM630抑制天疱疮引起的CB2、DSG3高表达及β-catenin低表达，可为天疱疮治疗提供新的治疗靶点和思路。

关键词: 大麻素受体, 自身免疫性疾病, 天疱疮, 桥粒黏蛋白, HaCaT细胞

Abstract:

Objective The aim of this study is to determine the effect of cannabinoid receptor (CB) 2 inhibitor on desmoglein 3 (DSG3) expression in HaCaT cells co-cultured with pemphigus serum. Methods Immunohistochemical staining was used to compare CB expression in pemphigus patients and normal individuals. Enzyme-linked immunosorbent assay (ELISA) was employed to quantify the concentration of CB2 in the serum of pemphigus patients and normal individuals. A correlation analysis was performed to examine the relationship between the serum CB2 and DSG of pemphigus patients. The CCK-8 assay was used to evaluate the inhibitory effect of AM630 on HaCaT cells, and the half-maximal inhibitory concentration (IC₅₀) value was utilized to determine the experimental concentration. Serum from normal individuals (negative control group) and pemphigus patients (pemphigus group) was co-cultured with HaCaT cells at a 1∶1 ratio. HaCaT cells cultured in complete medium were used as the control group. HaCaT cells in the pemphigus group treated with AM630 were employed as the AM630 group. Real-time polymerase chain reaction (PCR) and Western blot were conducted to assess the expression levels of CB2, DSG3, and β-catenin. Cell dissociation experiments were conducted to evaluate the effect of AM630 on the adhesion of HaCaT cells. Results Immunohistochemistry revealed significant differences in CB2 expression between pemphigus and normal mucosa (P<0.000 1), but no difference was found in CB1 expression. ELISA analysis revealed a statistically significant difference in the expression levels of CB2 in the serum between normal individuals and pemphigus patients (P<0.001). The expression of CB2 in the serum of pemphigus patients exhibited a significant positive correlation with that of DSG3 (r=0.831, P=0.003). The CCK-8 assay indicated that the IC₅₀ of AM630 on HaCaT cells was 0.55 μmol/L. Real-time PCR and Western blot showed that the expression levels of CB2 and DSG3 increased in the pemphigus group, while the expression level of β-catenin decreased compared with that in the AM630 groups (P<0.05). Conclusion CB2 is highly expressed in oral mucosal pemphigus. AM630 inhibits overexpression of CB2 and DSG3 and underexpression of β-catenin levels, which can provide new therapeutic targets for pemphigus.

Key words: cannabinoid receptor, autoimmune diseases, pemphigus, desmoglein, HaCaT cell

中图分类号:

R781.5

刘慧娟, 宋鹏, 侯亚丽, 霍晓, 秘利进, 刘春艳. 大麻素受体2抑制对口腔黏膜天疱疮棘层松解影响的研究[J]. 华西口腔医学杂志, 2025, 43(6): 829-836.

Liu Huijuan, Song Peng, Hou Yali, Huo Xiao, Mi Lijin, Liu Chunyan. Cannabinoid receptor 2 inhibition on acantholysis in oral mucosal pemphigus[J]. West China Journal of Stomatology, 2025, 43(6): 829-836.

图/表 7

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引物	序列
CB2	F：5’-TTGCCACTGAGGACCGAGGAC-3’
	R：5’-TCGCCCAAACAACAGGATGCC-3’
DSG3	F：5’-CTGTGCAGCAGCCTGGTAAGTC-3’
	R：5’-GCGAGCCACCGTTGAGAAGC-3’
β-catenin	F：5’-GCTCTTGTGCGTACTGTCCTTC-3’
	R：5’-TGGTGTCGGCTGGTCAGATG-3’
GAPDH	F：5’-TGGGTGTGAACCACGAGAA-3’
	R：5’-GGCATGGACTGTGGTCATGA-3’

大麻素受体2抑制对口腔黏膜天疱疮棘层松解影响的研究

Cannabinoid receptor 2 inhibition on acantholysis in oral mucosal pemphigus

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