摘要
目的探讨维生素D对特应性皮炎小鼠的保护作用及其作用机制。方法将清洁级的50只BALB/c雄性小鼠随机分为空白组、模型组、对照组和低、高剂量实验组,每组10只。除空白组外,其余组小鼠用2,4-二硝基氟苯(DNCB)进行建模,低、高剂量实验组小鼠建模后分别每天背部均匀涂抹5、10μg·kg^(-1)的1,25(OH)_(2)D_(3),对照组每天均匀涂抹氢化可的松溶液(1 mg·cm^(-2)),每天2次,连续给药10 d,空白组和模型组小鼠每天涂抹等量的0.9%NaCl。给药结束后对皮肤病变状况进行评估,眼眶采血并收集小鼠背部皮肤组织备用。以苏木精-伊红染色观察皮肤表皮厚度,以甲苯胺蓝染色法检测肥大细胞数量,以酶联免疫吸附测定(ELISA)法检测血清免疫球蛋白E(IgE)、白细胞介素(IL)-13、IL-4、IL-17表达水平,以流式细胞术检测血清Th1、Th2细胞比例,以免疫组化法检测背部皮肤组织组胺1型受体(HIR)、蛋白酶激活受体2(PAR2)、瞬时受体电位香草素1(TRPV1)表达情况。结果空白组、模型组、低剂量实验组、高剂量实验组和对照组小鼠的皮炎症状评分分别为0、(8.50±1.12)、(6.34±0.54)、(3.91±0.29)和(3.79±0.53)分,背部皮肤表皮厚度分别为(42.05±4.14)、(77.65±7.02)、(61.12±5.02)、(55.34±4.13)和(52.19±3.08)μm,背部组织肥大细胞数量分别为(4.67±0.27)、(32.16±2.49)、(25.23±2.07)、(18.13±1.46)和(15.37±1.29)number·mm^(-2),Th1/Th2细胞比值分别为2.76±0.28、0.36±0.06、1.02±0.18、2.05±0.14和2.07±0.29,HIR表达水平分别为0.05±0.00、0.21±0.02、0.16±0.02、0.10±0.01和0.08±0.01,PAR2表达水平分别为0.05±0.01、0.19±0.01、0.12±0.01、0.09±0.01和0.07±0.01,TRPV1表达水平分别为0.05±0.01、0.25±0.03、0.15±0.01、0.10±0.01和0.09±0.00;以上指标,低剂量实验组、高剂量实验组、对照组分别与模型组比较,高剂量实验组和低剂量实验组比较,在统计学上差异均有统计学意义(均P<0.05)。结论维生素D可通过调节Th1/Th2细胞平衡,减轻DNCB诱导的特应性皮炎,可能与HIR、PAR2介导的TRPV1痒信号通路有关。
Objective To investigate the protective effect of vitamin D on atopic dermatitis mice,and its mechanism.Methods Fifty male BALB/c mice in clean grade were randomly divided into blank group,model group,control group and experimental-L group,experimentalH group,10 rats in each group.Except for the blank group,mice in the other groups were modeled with 2,4-dinitrofluorobenzene(DNCB).After modeling,mice in experimental-L and experimental-H groups were evenly smeared with 5 and 10μg·kg~(-1)1,25(OH)_(2)D_(3)on their backs every day,respectively;control group were evenly coated with hydrocortisone solution(1 mg·cm^(-2))twice a day for 10 days,and the mice in blank group and model group were coated with the same amount of 0.9%NaCl every day.After the administration,the skin lesions were evaluated,and orbital blood was collected and the back skin tissues were collected for use.The epidermal thickness was observed by hematoxylin-eosin staining,and the number of mast cells was detected by toluidine blue staining.The expression levels of serum immunoglobulin E(IgE),interleukin(IL)-13,IL-4,IL-17 were detected by enzyme-linked immunosorbent assay(ELISA),and the proportion of serum Th1 and Th2 cells were detected by flow cytometry.The expression of histamine type 1 receptor(HIR),protease activating receptor 2(PAR2),transient receptor potential vanillin 1(TRPV1)in back skin was detected by immunohistochemistry.Results The symptom scores of mice in blank group,model group,experimental-L group,experimental-H group and control group were 0,(8.50±1.12),(6.34±0.54),(3.91±0.29)and(3.79±0.53)points;the epidermal thickness of the back skin were(42.05±4.14),(77.65±7.02),(61.12±5.02),(55.34±4.13)and(52.19±3.08)μm;the number of mast cells in back tissue were(4.67±0.27),(32.16±2.49),(25.23±2.07),(18.13±1.46)and(15.37±1.29)number·mm^(-2);Th1/Th2 cell ratios were 2.76±0.28,0.36±0.06,1.02±0.18,2.05±0.14 and 2.07±0.29,respectively;HIR expression levels were 0.05±0.00,0.21±0.02,0.16±0.02,0.10±0.01 and 0.08±0.01;the expression levels of PAR2 were 0.05±0.01,0.19±0.01,0.12±0.01,0.09±0.01 and 0.07±0.01;the expression levels of TRPV1 were 0.05±0.01,0.25±0.03,0.15±0.01,0.10±0.01 and 0.09±0.00,respectively.The above indicators,experimental-L group,experimental-H group and control group were compared with model group;experimental-H group compared with experimental-L group,the differences were all statistically significant(all P<0.05).Conclusion Vitamin D can reduce DNCB induced atopic dermatitis by regulating Th1/Th2 cell balance,which may be related to the TRPV1 oxygen signaling pathway mediated by HIR and PAR2.
作者
丁张军
吴志鹏
陈晓栋
DING Zhang-jun;WU Zhi-peng;CHEN Xiao-dong(Department of Dermatology,Affiliated Hospital of Nantong University,Nantong 226001,Jiangsu Province,China;Department of Dermatology,Dongtai People's Hospital,Yancheng 224200,Jiangsu Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2024年第6期884-888,共5页
The Chinese Journal of Clinical Pharmacology
