如意黑白散对白癜风模型小鼠皮损组织PD-1/PD-L1信号通路的影响  

Effect of Ruyi Heibai Power(如意黑白散)on PD-1/PD-L1 Signaling Pathway in Skin Lesion Tissue of Vitiligo Model Mice

作　　者：吴云莎 张星星[2] 王海丹[2] 吴玲[1] 张瑞娟 金玉[2] WU Yunsha;ZHANG Xingxing;WANG Haidan;WU Ling;ZHANG Ruijuan;JIN Yu(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing,210029;Jiangsu Provincial Hospital of Traditional Chinese Medicine)

机构地区：[1]南京中医药大学附属医院,江苏省南京市210029 [2]江苏省中医院

出　　处：《中医杂志》2024年第5期512-519,共8页Journal of Traditional Chinese Medicine

基　　金：江苏省中医院科技项目(Y2020CX19)。

摘　　要：目的探讨如意黑白散治疗白癜风的可能作用机制。方法将24只C57BL/6小鼠随机分为空白组,模型组及如意黑白散大、小剂量组,每组6只。除空白组外其余各组采用氢醌外涂构建白癜风动物模型。造模后如意黑白散大、小剂量组灌胃如意黑白散7.02、2.34 g/kg,空白组与模型组予10 ml/kg生理盐水灌胃,每日1次,各组均给药36天。给药结束后肉眼观察皮损情况,HE染色观察皮损组织黑色素含量,免疫组化测定皮损组织中CD3^(+)、CD8^(+)T细胞表达;免疫荧光染色测皮损组织中程序性死亡受体1(PD-1)表达;RT-PCR法检测皮损组织中细胞程序性死亡配体1(PD-L1)mRNA表达;ELISA法检测血清超氧化物歧酶(SOD)、肿瘤坏死因子(TNF-α)、酪氨酸酶(TYR)含量。结果与空白组比较,模型组小鼠皮肤苍白,皮损组织HE染色后镜下可见黑色素含量明显减少,皮损优良率降低,表皮和毛囊周围细胞黑色素颗粒明显减少,皮肤组织中CD3^(+)、CD8^(+)T细胞表达显著增加,PD-L1 mRNA、PD-1表达下降,TYR含量下降,SOD、TNF-α含量增加(P<0.05)。与模型组相比,如意黑白散大、小剂量组皮肤颜色加深,黑色素含量增加,皮损优良率升高,棘细胞层、基底细胞及毛囊黑色素颗粒增多,皮损处CD3^(+)、CD8^(+)T细胞表达减少,PD-L1 mRNA、PD-1表达增加,TYR含量上升,SOD、TNF-α含量下降(P<0.05)。与如意黑白散小剂量组比较,如意黑白散大剂量组造模区域皮损色素恢复面积更大,皮损优良率升高,棘细胞层及基底细胞以及毛囊黑色素颗粒增多,皮损处CD3^(+)、CD8^(+)T细胞表达减少,PD-L1 mRNA、PD-1表达增加,TYR含量上升,SOD、TNF-α含量下降(P<0.05)。结论如意黑白散可以使白癜风模型小鼠皮肤黑色素含量增多,其作用机制可能为激活皮损组织PD-1/PD-L1信号通路,从而抑制T细胞介导的自身免疫对黑色素细胞的破坏。Objective To investigate the possible mechanism of Ruyi Heibai Power(如意黑白散,RHP)in the treatment of vitiligo.Methods Twenty-four C57BL/6 mice were randomly divided into blank group,model group,high-dose and low-dose RHP groups,with 6 mice in each group.Model group,high-and low-dose RHP groups were all applied hydroquinone to establish vitiligo animal model.After modeling,High-and low-dose RHP groups were given 7.02 g/kg and 2.34 g/kg of RHP by gavage,respectively,while the blank group and model group were intragastrically given 10ml/kg of normal saline,once a day for 36 days.After administration,the skin lesions were observed with naked eye,and HE staining was used to observe the melanin content of the skin lesions.Immunohistochemistry was used to determine the expression of CD3^(+)and CD8^(+)T cells in skin tissue.Immunofluorescence staining was used to measure the expression of programmed death receptor 1(PD-1)in the skin lesion tissue.RT-PCR was used to detect programmed cell death ligand 1(PD-L1)mRNA expression.ELISA was used to detect serum superoxide dismutase(SOD),tumor necrosis factor(TNF-α),and tyrosinase(TYR)levels.Results Compared to those in the blank group,the skin of the mice in the model group was pale,and the melanin content was significantly reduced under the microscope after HE staining;the rate of excellent and good skin lesions decreased,and the melanin granules in the cells around the epidermis and hair follicles decreased significantly;the expression of CD3^(+) and CD8^(+)T cells in skin tissue increased significantly,and the expressions of PD-L1 mRNA and PD-1 decreased;the content of TYR decreased,while the content of SOD and TNF-αincreased(P<0.05).Compared to those in the model group,the skin color of high-and low-dose RHP groups were deepened,and the melanin content increased;the rate of excellent and good skin lesions increased,as well as the melanin granules in the spinous cell layer,basal cells and hair follicles;the expression of CD3^(+)and CD8^(+)T cells in the skin lesion

关 键 词：白癜风 黑色素 如意黑白散 程序性死亡受体1 程序性死亡配体1 T细胞 

分 类 号：R285.5[医药卫生—中药学]