基于ORAI1/NFAT信号轴探讨镇心安神方对1-氯-2,4-二硝基苯诱导特应性皮炎小鼠的作用及机制  

Effects and Mechanisms of Zhenxin Anshen Prescription on 1-Chloro-2,4-Dinitrobenzene-Induced Atopic Dermatitis Mice Based on ORAI1/NFAT Signaling Asix

作　　者：陈若曦 姜姗姗[2,3] 杨洁 杨晓宇[1,2] 徐蕊瑶 方琳 赵一丁 CHEN Ruoxi;JIANG Shanshan;YANG Jie;YANG Xiaoyu;XU Ruiyao;FANG Lin;ZHAO Yiding(Shaanxi University of Chinese Medicine,Xianyang 712046 Shaanxi,China;Shaanxi Academy of Traditional Chinese Medicine,Xi'an 710003 Shaanxi,China;Shaanxi Provincial Hospital of Traditional Chinese Medicine,Xi'an 710003 Shaanxi,China)

机构地区：[1]陕西中医药大学,陕西咸阳712046 [2]陕西省中医药研究院,陕西西安710003 [3]陕西省中医医院,陕西西安710003

出　　处：《中药新药与临床药理》2024年第2期168-173,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81704087,82274537);陕西省中医药管理局科研项目(2021-GJ-JC010)。

摘　　要：目的基于钙通道调节分子1(ORAI1)/T细胞核因子(NFAT)信号轴探讨镇心安神方(龙骨、牡蛎、淡竹叶、骨碎补、茯苓)对特应性皮炎(AD)小鼠的作用及机制。方法将36只BALB/c小鼠随机分为空白对照组、模型组、西替利嗪组(1.3 mg·kg^(-1))及镇心安神方组(36.36 g·kg^(-1)),每组9只。采用1-氯-2,4-二硝基苯(DNCB)诱导建立AD小鼠模型。灌胃给药,每日1次,连续给药2周。给药结束后,测定皮损面积,并对皮损严重程度进行评分;测定脾脏质量,计算脾脏指数;采用HE染色法观察皮损组织病理变化;ELISA法检测血清中白细胞介素4(IL-4)、IL-13及胸腺基质淋巴细胞生成素(TSLP)的含量;Western Blot法检测皮损组织中ORAI1、钙调磷酸酶A(CaN)、T细胞核因子2(NFAT2)蛋白表达水平。结果与空白对照组比较,模型组小鼠的皮损评分显著升高(P<0.01),皮损面积显著扩大(P<0.01);表皮层及真皮层厚度显著增加(P<0.01),表皮过度角化,棘层肥厚,真皮层可见嗜酸性粒细胞、淋巴细胞等炎性细胞浸润;脾脏指数及血清IL-4、IL-13、TSLP水平显著升高(P<0.01);皮损组织中CaN、NFAT2、ORAI1蛋白表达水平显著升高(P<0.01)。与模型组比较,镇心安神方组小鼠的皮损评分显著降低(P<0.01),皮损面积显著缩小(P<0.01);表皮层及真皮层厚度显著降低(P<0.01),表皮过度角化减轻,棘层轻度肥厚,真皮层有少量炎性细胞浸润;脾脏指数及血清IL-4、IL-13、TSLP水平显著降低(P<0.01);皮损组织中CaN、NFAT2、ORAI1蛋白表达水平显著降低(P<0.01)。结论镇心安神方能够改善DNCB诱导AD小鼠的皮肤病理损伤,可能与其抑制ORAI1、CaN及NFAT2蛋白表达,降低血清TSLP、IL-4、IL-13等2型炎症因子水平,通过免疫调节改善皮肤炎症及瘙痒有关。Objective To investigate the effects and mechanisms of Zhenxin Anshen Prescription(composed of Os Draconis,Ostreae Concha,Lophatheri Herba,Drynariae Rhizoma,Poria)on mice with atopic dermatitis(AD)based on the calcium channel regulator 1(ORAI1)/nuclear factor of T-cells(NFAT)signalling axis.Methods Thirty-six BALB/c mice were randomly divided into a blank control group,a model group,a Cetirizine group(1.3 mg·kg^(-1))and a Zhenxin Anshen Prescription group(36.36 g·kg^(-1)),with nine mice in each group.AD mouse model was established using 1-chloro-2,4-dinitrobenzene(DNCB)induction.The drug was administered by gavage once a day for 2 weeks.At the end of drug administration,the area of skin lesions was measured and the severity of skin lesions was scored;spleen mass was measured and spleen index was calculated;pathological changes of skin lesion tissues were observed by HE staining;interleukin 4(IL-4),IL-13 and thymic stromal lymphopoietin(TSLP)in serum were detected by ELISA;and the protein expression levels of ORAI1,calmodulin phosphatase A(CaN)and nuclear factor of T cells 2(NFAT2)were detected by Western Blot.Results Compared with the blank control group,the skin lesion score of mice in the model group was significantly increased(P<0.01),the skin lesion area was significantly enlarged(P<0.01);the thickness of the epidermis and dermis were significantly increased(P<0.01),hyperkeratosis of the epidermis,hypertrophy of the stratum spinosum,and infiltration of inflammatory cells such as eosinophils and lymphocytes can be seen in the dermis;the splenic index and serum IL-4,IL-13,TSLP levels were significantly increased(P<0.01);protein expression levels of CaN,NFAT2,ORAI1 were significantly increased in the skin lesion tissues(P<0.01).Compared with the model group,the dermatitis score of mice in the Zhenxin Anshen Prescription group was significantly decreased(P<0.01),the lesion area was significantly reduced(P<0.01),the epidermal and dermal thicknesses(P<0.01),the hyperkeratosis of epidermis was alleviated,the spino

关 键 词：镇心安神方 特应性皮炎 ORAI1/NFAT信号轴 免疫调节 小鼠 

分 类 号：R285.5[医药卫生—中药学]