脂肪来源间充质干细胞对免疫性脱发的治疗作用及机制研究

Effects of adipose derived mesenchymal stem cells in the treatment of immune alopecia and related mechanism

目的探讨脂肪来源间充质干细胞(ADSCs)对免疫性脱发的影响。方法采用Transwell小室(0.4μm)将人源ADSCs和毛囊真皮乳头细胞(DPCs)共培养,通过CCK-8法观察共培养后DPCs增殖能力的变化,并通过实时荧光PCR检测共培养DPCs诱导能力的变化。将40只雄性C57BL/6小鼠随机分成4组:空白对照组、免疫性脱发组、脂肪干细胞治疗组、糖皮质激素治疗组。通过建立小鼠免疫性脱发模型,观察各组小鼠脱毛区皮肤颜色变化和新生毛发生长状况,计算各组小鼠新生毛发长度、质量,通过苏木精-伊红染色(H-E)观察各组小鼠毛囊数量和形态差异,通过免疫组化观察毛囊周围炎症因子细胞间黏附分子-1(ICAM-1)和内皮细胞黏附分子-1(ELAM-1)水平和免疫因子CD4^(+)、CD8^(+)水平。结果研究发现ADSCs可以有效促进DPCs的增殖能力,增强DPCs的分泌功能(P<0.01)及DPCs诱导毛囊形成的能力,促进毛囊进入生长期。在免疫性脱发小鼠模型中,经ADSCs混合培养基注射治疗,小鼠毛发质量增加,毛发生长情况改善(P<0.05),H-E及免疫组化结果显示ADSCs可以下调毛囊周围的免疫功能,降低局部炎症因子水平。结论局部ADSCs混合培养基注射可以治疗免疫性脱发,可能与调节局部免疫微环境及抑制炎症因子对毛囊的破坏有关。提示ADSCs局部应用可以作为难治性免疫性脱发治疗的新思路。

Objective To investigate the effect of adipose derived mesenchymal stem cells(ADSCs)on immune alopecia.Methods Human ADSCs and hair follicle dermal papilla cells(DPCs)were co-cultured in Transwell chambers(0.4μm).The proliferation of DPCs after co-culture was observed by CCK-8 assay,and the induction ability of DPCs was detected by real-time fluorescence PCR.Furthermore,40 male C57BL/6 mice were randomly divided into four groups:a blank control group,an immune alopecia group,an adipose stem cell treatment group,and a glucocorticoid treatment group.An immune alopecia model of mice was established to observe the changes of skin color and the growth of new hair in the depilation area of mice in each group.The length and quality of new hair in each group were calculated.The number and morphological differences of hair follicles in each group were observed by H-E staining.The levels of inflammatory cytokines intercellular adhesion molecule-1(ICAM-1)and endothelial leukocyte adhesion molecule-1(ELAM-1)and immune cytokines CD4^(+)and CD8^(+)around hair follicles were observed by immunohistochemistry.Results The results showed that ADSCs effectively promoted the proliferation ability of DPCs(P<0.01),enhance the secretion function of DPCs(P<0.01),strengthen the ability of DPCs to induce hair follicle formation,and promote hair follicle into the growth phase.In the mouse model of immune alopecia,injection of ADSCs increased hair mass(P<0.05)and improved hair growth(P<0.05).According to H-E staining and immunohistochemistry,ADSCs down-regulated the immune function around the hair follicle and reduced the level of local inflammatory factors.Conclusions Local injection of ADSCs can treat immune alopecia,which may be related to regulating local immune microenvironment and inhibiting the damage of inflammatory factors to hair follicles.It is suggested that local application of ADSCs may be a new thought for the treatment of refractory immune alopecia.