Operating Room, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China.
Wound Treatment Department, TCM-Integrated Hospital of Southern Medical University, Guangzhou, Guangdong Province, China.
Acta Histochem. 2021 Oct;123(7):151789. doi: 10.1016/j.acthis.2021.151789. Epub 2021 Sep 22.
The mechanisms underlying the role of mast cells in wound healing have not been thoroughly studied, and even fewer data are available on studies related to mast cells in the skin of patients with type 2 diabetes mellitus (T2DM). Therefore, this study aims to explore the transcriptional characteristics of mast cells at the single-cell level in patients with T2DM and provide experimental data for studying mast cell behaviors under abnormal glucose metabolism.
Two patients with T2DM and one trauma patient without diabetes were enrolled. Samples were derived from skin tissue resected at the time of surgery and were isolated by single cell capture technology on BD platform to prepare single cell cDNA library. Seurat was used to process raw reads and analyze data downstream of single-cell RNA sequencing, including removal of low-quality cells, identification of cell clusters at the single-cell level, and screening for differential genes with fold change > 1.5 and p < 0.05 by two-sided t-test. We performed single-cell RNA sequencing on skin tissues of T2DM patients and non-diabetics and identified the cell cluster of skin, single-cell subsets, and transcriptional characteristics of mast cells at a single-cell level. Meanwhile, gene set enrichment(GSEA) analysis was performed on the differentially expressed genes.
A total of 8888 cells were obtained from skin tissue. Clustering analysis revealed eight-cell clusters, identified as smooth muscle cells, dendritic cells, mast cells, and T cells, respectively. Cluster 6 was identified as mast cells with the marker genes TPSAB1, CPA3, TPSB2, MS4A2,KIT, etc., which accounting for 2.7% of the total cell number.Compared with the control group, the genes highly expressed in MCs from T2DM patients, include ADH1C, PAXIP1, HAS1, ARG1, etc., and the low expression genes include PHACTR2, GGA1, RASSF2, etc. GSEA analysis suggested that the signal pathways of MCS in T2DM patients included VEGF signaling pathway, Fc gamma R-mediated phagocytosis, the B cell receptor signaling pathway, natural killer cell-mediated cytotoxicity.
The characteristic genes of MCs in the skin tissues of T2DM patients were described at the single-cell level. These genes and enriched signaling pathways provide a theoretical basis and data support for further researches on dermatopathy in patients with diabetes mellitus.
尽管mast 细胞（肥大细胞）在伤口愈合中的作用机制尚未得到深入研究，但有关 2 型糖尿病（T2DM）患者皮肤中 mast 细胞的研究数据则更少。因此，本研究旨在探索 T2DM 患者的mast 细胞在单细胞水平上的转录特征，并为研究异常葡萄糖代谢下 mast 细胞行为提供实验数据。
纳入 2 例 T2DM 患者和 1 例无糖尿病创伤患者。标本取自手术切除的皮肤组织，采用 BD 平台单细胞捕获技术分离，制备单细胞 cDNA 文库。使用 Seurat 处理原始读数，并对单细胞 RNA 测序的下游数据进行分析，包括去除低质量细胞、单细胞水平上鉴定细胞簇，以及通过双侧 t 检验筛选 fold change > 1.5 和 p < 0.05 的差异基因。我们对 T2DM 患者和非糖尿病患者的皮肤组织进行了单细胞 RNA 测序，鉴定了皮肤、单细胞亚群和mast 细胞在单细胞水平上的转录特征。同时，对差异表达基因进行了基因集富集（GSEA）分析。
从皮肤组织中获得了 8888 个细胞。聚类分析显示了 8 个细胞簇，分别鉴定为平滑肌细胞、树突状细胞、mast 细胞和 T 细胞。簇 6 被鉴定为 mast 细胞，其标记基因包括 TPSAB1、CPA3、TPSB2、MS4A2、KIT 等，占总细胞数的 2.7%。与对照组相比，T2DM 患者 mast 细胞中高表达的基因包括 ADH1C、PAXIP1、HAS1、ARG1 等，低表达的基因包括 PHACTR2、GGA1、RASSF2 等。GSEA 分析表明，T2DM 患者 MCS 的信号通路包括 VEGF 信号通路、FcγR 介导的吞噬作用、B 细胞受体信号通路、自然杀伤细胞介导的细胞毒性等。
描述了 T2DM 患者皮肤组织中 mast 细胞的特征基因。这些基因和富集的信号通路为进一步研究糖尿病患者的皮肤病变提供了理论依据和数据支持。
