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目的探讨复方黄柏液涂剂对人脂肪间充质干细胞(A-MSC)免疫调控能力的影响情况。方法从人皮下脂肪样本分离间充质干细胞,观察形态学改变和细胞表型,用A-MSC完全培养基按梯度稀释后进行流式细胞术凋亡检测,按1∶100稀释后,用CCK-8法检测复方黄柏液涂剂处理24h后的A-MSC对淋巴细胞增殖能力的影响,实时定量PCR法检测复方黄柏液涂剂处理后免疫调控相关基因在A-MSC中的表达情况。结果所得A-MSC具有典型成纤维细胞形态和显著增殖能力,阳性表达CD29,CD44,CD73,CD90和CD105,不表达CD31,CD45和CD133等血细胞标记,复合MSC国际标准。予低浓度复方黄柏液涂剂处理后,A-MSC对淋巴细胞增殖的抑制作用明显增强,且A-MSC中细胞免疫抑制相关基因表达增加,促炎相关基因表达减弱。结论复方黄柏液涂剂可通过加强脂肪间充质干细胞免疫抑制能力影响创伤局部炎性微环境。
Objective To investigate the effect of compound HuangPu liquid coating agent on immune regulation of human adipose-derived mesenchymal stem cells (A-MSCs). Methods Mesenchymal stem cells (MSCs) were isolated from human subcutaneous adipose tissue. Morphological changes and cell phenotype were observed. Apoptosis was detected by flow cytometry with gradient dilution of A-MSC complete medium. After diluted 1: 100, -8 method was used to detect the effect of A-MSCs on the proliferation of lymphocytes treated with Cortex Phellodendri liquid agent for 24 hours. The expression of immune-related genes in A-MSC was detected by real-time quantitative PCR method. Results The obtained A-MSCs showed typical fibroblast morphology and significant proliferative capacity. The positive expression of CD29, CD44, CD73, CD90 and CD105, but not the expression of CD31, CD45 and CD133 and other international standards of MSC. After being treated with low concentration of Cortex Phellodendri, the inhibitory effect of A-MSC on lymphocyte proliferation was significantly enhanced, and the expression of cellular immunosuppression-related genes increased and the expression of proinflammatory gene decreased in A-MSC. Conclusion The compound cortex phellodendri liquidum can affect the local inflammatory microenvironment of wounds by strengthening the immunosuppressive capacity of adipose-derived mesenchymal stem cells.