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广州中医药大学第七临床医学院,广东 深圳 518133
广州中医药大学附属宝安中医院,广东 深圳 518133
广州中医药大学第一附属医院
范思敏,广州中医药大学硕士研究生。E-mail:1055698716@qq.com
✉李晶晶,副主任医师。E-mail:lijingjing85@gzucm.edu.cn
收稿:2025-01-16,
网络首发:2026-04-27,
纸质出版:2026-06-12
移动端阅览
范思敏, 李晶晶, 曾婧纯, 等. 基于皮肤组织转录组测序探讨火针对氢醌诱导白癜风模型的作用机制[J]. 中国针灸, 2026,46(6):987-1000.
FAN Simin, LI Jingjing, ZENG Jingchun, et al. Effect mechanism of fire needle therapy on vitiligo induced by hydroquinone in modeled mice based on skin tissue transcriptome sequencing[J]. Chinese Acupuncture & Moxibustion, 2026, 46(6): 987-1000.
范思敏, 李晶晶, 曾婧纯, 等. 基于皮肤组织转录组测序探讨火针对氢醌诱导白癜风模型的作用机制[J]. 中国针灸, 2026,46(6):987-1000. DOI: 10.13703/j.0255-2930.20250116-0002.
FAN Simin, LI Jingjing, ZENG Jingchun, et al. Effect mechanism of fire needle therapy on vitiligo induced by hydroquinone in modeled mice based on skin tissue transcriptome sequencing[J]. Chinese Acupuncture & Moxibustion, 2026, 46(6): 987-1000. DOI: 10.13703/j.0255-2930.20250116-0002.
目的:
2
基于转录组测序探讨火针干预氢醌诱导的白癜风的可能作用机制。
方法:
2
将18只C57BL/6小鼠随机分为空白组、模型组、火针组,每组6只。模型组和火针组采用氢醌脱色法制备白癜风小鼠模型。造模成功后,火针组于白色皮损处进行火针干预,每周1次,共干预4周。观察干预前后小鼠皮肤毛发脱色情况及皮肤毛发脱色评分,HE染色观察小鼠皮肤形态及黑素颗粒情况。各组随机取3只小鼠的局部白斑处皮肤组织采用RNA-seq技术筛选小鼠皮肤组织中差异表达的基因,进行生物信息学分析。并采用实时荧光定量PCR法检测部分差异基因的mRNA表达。
结果:
2
与空白组比较,模型组小鼠皮损区表皮层和真皮层增厚,皮肤毛发脱色评分升高(
P
<
0.05),表皮皮肤含黑素细胞毛囊数、基底层黑素细胞数、含黑素颗粒表皮细胞数减少(
P
<
0.05);与模型组比较,火针组小鼠皮肤毛发脱色评分降低(
P
<
0.05),表皮皮
肤含黑素细胞毛囊数、基底层黑素细胞数、含黑素颗粒表皮细胞数增加(
P
<
0.05)。与空白组比较,模型组共筛选出1 291个差异表达基因(包括985个上调、306个下调),基因本体(GO)功能富集分析主要在角质化、角化包膜,京都基因与基因组百科全书(KEGG)代谢通路分析主要富集在病毒蛋白与细胞因子和细胞因子受体的相互作用;与模型组比较,火针组共筛选出306个差异表达基因,GO功能富集分析主要在角化包膜、皮肤屏障的建立,KEGG代谢通路分析主要富集在细胞因子和细胞因子受体的相互作用。维基通路(WikiPathways)富集分析发现,火针组氧化应激和氧化还原途径显著。通过基因集富集分析(GSEA)显示,河马(Hippo)信号通路、过氧化物酶体增殖物激活受体(PPAR)信号通路被显著激活。与空白组比较,模型组小鼠皮肤组织超氧化物歧化酶3(SOD3)、过氧化物酶体增殖物激活受体γ(PPARγ)、醌氧化还原酶1(NQO1)、单胺氧化酶(MAO)、白细胞介素18(IL-18)、白细胞介素1家族成员8(IL-1F8)、白细胞介素1家族成员9(IL-1F9)、C-C基序趋化因子配体8(CCL8)、丝氨酸/苏氨酸蛋白激酶4(MST1)、MOB激酶激活因子1A(MOB1A)mRNA表达升高(
P
<
0.05),谷胱甘肽S-转移酶α 3(GSTA3)、谷胱甘肽S-转移酶α 4(GSTA4)、γ-谷氨酰环转移酶(GGCT)、Wnt家族成员3A(WNT3A)、酪氨酸酶相关蛋白1(TYRP1)、过氧化氢酶(CAT)、叉头框蛋白P3(FOXP3)mRNA表达降低(
P
<
0.05);与模型组比较,火针组小鼠皮肤组织SOD3、PPARγ、NQO1、MAO、IL-18、IL-1F8、IL-1F9、CCL8、MST1、MOB1A mRNA表达降低(
P
<
0.05),GSTA3、GSTA4、GGCT、WNT3A、TYRP1、CAT、FOXP3 mRNA表达升高(
P
<
0.05)。
结论:
2
火针干预能够改善氢醌诱导的白癜风小鼠皮肤脱色症状,其作用机制可能涉及调节Hippo信号通路和PPAR信号通路,从而减轻氧化应激、减少炎症反应,并调控免疫系统,最终促进黑素细胞恢复,减少白斑面积。
Objective
2
To explore the underlying effect mechanism of fire needle therapy on hydroquinone-induced vitiligo based on transcriptome sequencing.
Methods
2
Eighteen C57BL/6 mice were randomly divided into a blank group
a model group
and a fire needle group
with 6 mice in each group. The vitiligo mouse model was established in the model and fire needle groups using the hydroquinone depigmentation method. After successful modeling
in the fire needle group
fire needle therapy was operated on the white skin lesions
once a week and for 4 consecutive weeks. The skin and hair depigmentation conditions and depigmentation scores were observed before and after intervention. HE staining was used to observe skin morphology and melanin granules. Skin tissue from the local white lesions of 3 randomly selected mice in each group was subjected to RNA-seq to screen differentially expressed genes in skin tissue
followed by bioinformatics analysis.The mRNA expression of the partial differentially expressed genes was detected by real-time fluorescent quantitative PCR.
Results
2
Compared with the blank group
in the model group
the epidermis and dermis were thickened
and the depigmentation score of the skin and hair was higher (
P
<
0.05)
and the number of melanocytes in the epidermis and hair follicles
the number of melanocytes in the basal layer
and the number of epidermal cells containing melanin granules were decreased (
P
<
0.05). Compared with the model group
the depigmentation score of the skin and hair in the fire needle group was decreased (
P
<
0.05)
and the number of melanocytes in the epidermis and hair follicles
the number of melanocytes in the basal layer
and the number of epidermal cells containing melanin granules were increased (
P
<
0.05). In comparison with the blank group
1 291 differentially expressed genes were screened in the model group (including 985 up-regulated genes and 306 down-regulated genes)
the functional enrichment analysis of gene ontology (GO) primarily focused on keratinization and keratinized envelope
and the metabolic pathway analysis from the Kyoto encyclopedia of genes and genomes (KEGG)was mainly enriched in the interactions between viral proteins and cytokines/cytokine receptors. When compared with the model group
306 differentially expressed genes were screened in the fire needle group
and the GO enrichment was in keratinized envelope and the establishment of skin barrier
and the KEGG metabolic pathway analysis was enriched in the interaction between cytokines and cytokine receptors. Through WikiPathways enrichment analysis
fire needle therapy significantly exerted its effect through the oxidative stress and redox pathways. The gene set enrichment analysis (GSEA)showed that the Hippo signaling pathway and peroxiso
me proliferator-activated receptor (PPAR) signaling pathway were significantly activated. Compared with the blank group
in the model group
the mRNA expression was elevated for superoxide dismutase 3 (SOD3)
peroxisome proliferator-activated receptor γ (PPARγ)
quinone oxidoreductase 1(NQO1)
monoamine oxidase (MAO)
interleukin 18 (IL-18)
interleukin 1 family member 8 (IL-1F8)
interleukin 1 family member 9 (IL-1F9)
C-C motif chemokine ligand 8 (CCL8)
serine/threonine protein kinase 4 (MST1)
and MOB kinase activator 1A (MOB1A) in the skin tissue (
P
<
0.05)
and the mRNA expression was reduced for glutathione S-transferase alpha 3 (GSTA3)
glutathione S-transferase alpha 4 (GSTA4)
gamma-glutamyl cyclotransferase (GGCT)
Wnt family member 3A (WNT3A)
tyrosinase-related protein 1 (TYRP1)
catalase (CAT)
and forkhead box P3 (FOXP3) (
P
<
0.05). Compared with the model group
the fire needle group showed the decrease in the mRNA expression of SOD3
PPARγ
NQO1
MAO
IL-18
IL-1F8
IL-1F9
CCL8
MST1 and MOB1A in the skin tissue (
P
<
0.05)
and the increase in the mRNA expression of GSTA3
GSTA4
GGCT
WNT3A
TYRP1
CAT and FOXP3 (
P
<
0.05).
Conclusion
2
Fire needling therapy can attenuate the depigmentation of skin in vitiligo induced by hydroquinone in mice. Its effect mechanism may involve the regulation of Hippo signaling pathways and PPAR signaling pathways
thereby mitigating oxidative stress and inflammatory response
and regulating the immune system
ultimately promoting the recovery of melanocytes and reducing the size of white patches.
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