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 effects of compound fluid of Cortex Phellodendri (CFCP)on immunomodulatory ability of human adipose derived mesenchymal stem cell(A-MSC). Methods Mesenchymal stem cells(MSCs)were isolated from human subcutaneous adipose tissue and observe its morphological characterization and immune phenotype. Apoptosis analysis was performed after gradient dilution using A-MSC complete medium using flow cytometry. After treatment by 1:100 diluted CFCP, CCK-8 method was used to determine the influence of A-MSC on lymphocyte proliferation. The expression of immunomodulation related genes in CFCP treated A-MSC were investigated by real time PCR. Results A-MSC have a typical fibroblastic morphology and strong proliferation capability, positive expression of CD29, CD44, CD73, CD105, do not express CD31, CD45, CD133, which is consistent with the international standard of MSC. After low concentrationc CFCP treatment, the inhibition of A-MSC on lymphocyte proliferation enhanced. And CFCP treated A-MSC express more immune cells inhibit related gene and less proinflammatory gene. Conclusion CFCP can strengthen the immune suppression ability of A-MSC to influence the wound inflammatory microenvironment.