目的：探究女阴硬化性苔藓（vulva lichen sclerosus，VLS）给患者生活质量造成的影响及带来的疾病负担；探究成纤维细胞中阿斯波林（Asporin，ASPN）在VLS发病中潜在的作用机制。
方法：
1. 回顾性研究纳入607例VLS 患者，根据发病年龄将其分为青春期前组（96例）、生育期组（400例）和绝经后组（111例）。采集患者的一般情况、生活质量和疾病负担的信息，资料采用皮肤病生活质量指数（the Dermatology Life Quality Index, DLQI）问卷和临床检查相结合的方法收集。
2. 通过单细胞转录组测序技术对比VLS患者皮损部位皮肤组织及正常外阴皮肤组织转录组信息，分析VLS中成纤维细胞的核心作用以及和相关细胞的通讯功能。
3. 通过免疫荧光、RT-qPCR、Western Blot、Transwell等实验探究ASPN在成纤维细胞中细胞外基质合成及CD8+ T细胞趋化方面的作用。
结果：
1. VLS严重影响患者生活质量，整体患者人群的DLQI评分中位数为6。瘙痒是影响VLS患者生活质量的主要症状，约93%，瘙痒评分中位数为5。患者人群夜间瘙痒的发生率为49.7%。82.8%的VLS患者性生活受到影响，12.8%的VLS患者因患病而过度清洁外阴。同时VLS也带给患者很大的疾病负担。患者既往治疗费用中位数为2000元人民币。与生育期组和绝经后组患者相比，青春期前组患者受瘙痒的影响少，仅20.7%的患者夜间瘙痒影响睡眠。青春期前组患者生活质量受累较轻，DLQI评分中位数为3。
2. 通过单细胞转录组测序技术将皮肤组织共分为12个细胞亚群，与健康对照相比，VLS皮损组织中淋巴内皮细胞、施万细胞和T细胞数量比例升高，成纤维细胞、巨噬树突细胞、肥大细胞和黑素细胞比例减少。VLS中成纤维细胞差异基因富集在胶原合成、细胞外基质等通路。将成纤维细胞分为分泌乳头型、分泌网状型、促炎型和间充质型4种亚型，发现仅间充质型成纤维细胞亚群比例在VLS中显著升高。部分成纤维细胞亚群的差异基因富集在细胞外基质合成相关通路，以及MHC蛋白复合物识别相关通路。疾病组各成纤维细胞亚群之间的细胞间通讯增强，并且CD8+ T细胞与成纤维细胞亚群在COLLAGE等通路上的通讯增强。

3. ASPN
在VLS组织的成纤维细胞中表达增多，促进成纤维细胞中Ⅰ型、Ⅲ型、Ⅳ型胶原和FGF2的合成，抑制弹性蛋白的表达，并通过分泌CXCL9、CXCL11等趋化因子促进CD8+ T细胞的迁移。
结论：
1. VLS 给患者带来了巨大的生理、心理和经济负担。
2. VLS中成纤维细胞可以分为4种亚型，部分亚群促进细胞外基质的形成；与CD8+ T细胞的通讯增强。
3. VLS中ASPN可以促进成纤维细胞合成细胞外基质并通过分泌趋化因子促进对CD8+ T细胞的趋化。

Objective: To investigate the influence of vulva lichen sclerosus (VLS) on the quality of life and the burden of disease, and to explore the potential role of Asporin (ASPN) in the pathogenesis of VLS.
Method:
1. The retrospective study included 607 patients with VLS. They were divided into prepubertal group (96 cases) , reproductive group (400 cases) and postmenopausal group (111 cases) according to the age of onset. The Dermatology Life Quality Index (DLQI) and clinical examination were used to collect the information of patients, such as general conditions, quality of life and disease burden.
2. scRNA-seq was used to compare the skin lesions of patients with VLS and normal vulvar skin tissues to analyze the core role of fibroblasts in VLS and the function of communication with related cells.
3. Immunofluorescence, RT-qPCR, Western Blot and Transwell techniques were used to explore the role of ASPN in extracellular matrix synthesis and CD8+ T cell chemotaxis in fibroblasts.
Result:
1. VLS seriously affected the quality of life of patients, and the median score of the Dermatology Life Quality Index (DLQI) of the whole population was 6. Itching is the main symptom affecting the quality of life of patients with VLS, about 93%, and the median pruritus score is 5. The incidence of night itching in the overall patient population was 49.6%. 82.8% of VLS patients' sex life was affected, and 12.8% of VLS patients excessively cleaned the vulva because of the disease. At the same time, VLS also brings a great burden of disease to patients. The median cost of previous treatment was 2000 RMB. Compared with the childbearing group and the postmenopausal group, patients in the prepubertal group were less affected by pruritus, and only 20.7% of the patients with night itching affected sleep. The quality of life in the prepubertal group was less affected, and the median DLQI score was 3.
2. The skin tissue was divided into 12 cell subsets by scRNA-aeq. Compared with healthy controls, the proportion of lymphatic endothelial cells, Schwann cells and T cells in VLS lesions increased, while the proportion of fibroblasts, macrophages-dendritic cells, mast cells and melanocytes decreased. The differential genes of fibroblasts composed of VLS are enriched in the pathways of collagen synthesis and extracellular matrix. Fibroblasts were divided into four subtypes: Secretory-papillary, Secretory-reticular, Pro-inflammatory and Mesenchymal. It was found that only the proportion of Mesenchymal fibroblasts subsets increased significantly in VLS. The differential genes of the four fibroblast subsets were enriched in the collagen synthesis pathway, and the differential genes of Pro-inflammatory, Mesenchymal and Secretory-reticular fibroblasts were enriched in the MHC protein complex recognition pathway. In the disease group, the intercellular communication between fibroblast subsets was enhanced, and the communication between CD8+ T cells and fibroblast subsets in COLLAGE and other pathways was enhanced.
3. The expression of ASPN was increased in fibroblasts of VLS tissue. ASPN promoted the synthesis of collagen I, Ⅲ, IV and FGF2 in fibroblasts, inhibited the expression of elastin. It promoted the migration of CD8+ T cells by secreting chemokines such as CXCL9 and CXCL11.
Conclusion:
1. VLS brings huge physiological, psychological and economic burden to patients.
2. Fibroblasts in VLS can be divided into four subtypes, all of which promote the formation of extracellular matrix and enhance communication with CD8+ T cells.
3. ASPN in VLS can promote fibroblasts to synthesize extracellular matrix and promote the chemotaxis of CD8+T cells by secreting chemokines.