利用微小毛发模型验证α-倒捻子素在毛囊中的抗氧化特性的研究

Using a micro hair model to verify the antioxidant properties of α-mangostin in hair follicles

目的探究药物α-倒捻子素(αMG)在毛囊组织中的抗氧化能力,推测其保护毛发免受自由基伤害、减缓毛发色素脱失的潜力。方法基于固-液培养方法建立人头皮毛囊微小毛发模型,经2%过氧化氢(H_(2)O_(2))预刺激后,加入浓度分别为10μmol/L、40μmol/L的αMG处理孵育。检测经H_(2)O_(2)诱导及αMG的处理后,毛发中活性氧(ROS)的含量与黑色素颗粒产生的位置与数量,以及毛发中的细胞凋亡水平。组间比较采用单因素方差分析,多组间两两比较采用LSD-t检验。结果40μmol/LαMG降低了毛发中的ROS水平,H_(2)O_(2)组、H_(2)O_(2)+40μmol/LαMG组的ROS水平分别为空白对照组的(1.44±0.05)、(1.07±0.09)倍。因强氧化剂导致的毛干部黑色素颗粒增加的程度受到抑制,通过抗氧化的特性抑制毛囊黑色素颗粒的分化与迁移,培养7天异位黑色素颗粒数量从H_(2)O_(2)组的(11.3±2.8)个下降至H_(2)O_(2)+40μmol/LαMG组的(4.1±3.2)个。αMG还表现出抑制毛发的细胞凋亡的作用,细胞凋亡荧光染色强度相较对照组从H_(2)O_(2)组的(1.79±0.13)倍下降至H_(2)O_(2)+40μmol/LαMG组(0.83±0.34)倍,其抗氧化水平与强抗氧化剂N-乙酰半胱氨酸相当。结论微小毛发模型可作为毛发黑色素细胞分化迁移及色素改变的研究模型,验证药物αMG的抗氧化作用可以减少H_(2)O_(2)诱导的毛囊中ROS积累和细胞凋亡,抑制毛囊在应激状态(如强氧化剂作用)下的黑色素颗粒异位分泌。

Objective To explore the antioxidant capacity of the drugα-mangostin(αMG)in hair follicle tissue,and speculate on its potential to protect hair from free radical damage and slow down hair pigmentation loss.Methods A micro-hair model of human scalp hair follicles was established based on the solid-liquid culture method.After pre-stimulation with 2%hydrogen peroxide(H_(2)O_(2)),αMG with concentrations of 10μmol/L and 40μmol/L was added for incubation.After H_(2)O_(2) induction andαMG treatment,the content of reactive oxygen species(ROS)in the hair,the location and number of melanin granules produced,and the level of cell apoptosis in the hair were detected.One-way analysis of variance was used for comparison between groups,the LSD-t test was used for pairwise comparison among multiple groups.Results 40μmol/L+αMG significantly decreased the ROS levels in hair.ROS levels in the H_(2)O_(2) group and H_(2)O_(2)+40μmol/LαMG group were(1.44±0.05)and(1.07±0.09)times that of the blank control group,respectively.The increased in the production of melanin particles in the hair shaft caused by strong oxidants is inhibited,which in turn hinders the differentiation and migration of hair follicle melanin particles due to its antioxidant properties.The number of ectopic melanin particles in the H_(2)O_(2) group increased from(11.3±2.8)to(4.1±3.2)in the H_(2)O_(2)+40μmol/LαMG group after 7 days of culture.αMG also demonstrates the inhibition of hair cell apoptosis.The fluorescence staining intensity of apoptosis decreased from(1.79±0.13)in the H_(2)O_(2) group to(0.83±0.34)in the H_(2)O_(2)+40μmol/LαMG group when compared to the control group.Its antioxidant efficacy is comparable to that of the potent antioxidant N-acetylcysteine.Conclusion The micro-hair model can be used as a research model for hair melanocyte differentiation,migration and pigment changes.It verifies that the antioxidant effect of the drugαMG can significantly reduce the accumulation of ROS and cell apoptosis in hair follicles induced by H_(2)O_(2),inhibit the ectopic secretion of melanin granules in hair follicles under stress conditions,such as the action of strong oxidants.