树突状细胞在咪喹莫特诱导的银屑病样小鼠模型中的表达及凉血解毒方的干预作用

doi:10.3969/j.issn.1006-7795.2017.02.019

首都医科大学学报 ›› 2017, Vol. 38 ›› Issue (2): 249-259.doi: 10.3969/j.issn.1006-7795.2017.02.019

树突状细胞在咪喹莫特诱导的银屑病样小鼠模型中的表达及凉血解毒方的干预作用

王明星^1,2, 王燕^2,3, 赵京霞^1,2, 底婷婷^2,3, 阮智通^1,2, 蒙玉娇^1,2, 谢湘江^2,3, 张璐², 林燕², 王宁², 李萍²

1. 首都医科大学附属北京中医医院, 北京 100010;
2. 银屑病中医临床基础研究北京市重点实验室, 北京市中医研究所, 北京 100010;
3. 北京中医药大学, 北京 100027

收稿日期:2016-12-07 出版日期:2017-03-21 发布日期:2017-04-17
通讯作者: 李萍,E-mail:liping411@126.com E-mail:liping411@126.com
基金资助:
国家自然科学基金青年科学基金（81403410），北京市科技计划课题“十病十药”研发（Z141100002214015）

Dendritic cells on mice psoriasis-like lesions induced by imiquimod and the effects of Liangxue Jiedu decoction

Wang Mingxing^1,2, Wang Yan^2,3, Zhao Jingxia^1,2, Di Tingting^2,3, Ruan Zhitong^1,2, Meng Yujiao^1,2, Xie Xiangjiang^2,3, Zhang Lu², Lin Yan², Wang Ning², Li Ping²

1. Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, China;
2. Beijing Key Laboratory of Clinic and Basic Research with Traditional Chinese Medicine on Psoriasis, Beijing Institute of Traditional Chinese Medicine, Beijing 100010, China;
3. Beijing University of Chinese Medicine, Beijing 100027, China

Received:2016-12-07 Online:2017-03-21 Published:2017-04-17
Supported by:
This study was supported by National Natural Science Foundation of China Youth Fund (81403410), Beijing Project of Science and Technology Plan (Z141100002214015)

摘要/Abstract

摘要： 目的观察凉血解毒方对咪喹莫特（imiquimod，IMQ）诱导的银屑病样小鼠模型中活化的树突状细胞（dendritic cells，DCs）的干预作用。方法将小鼠采用数字表法随机分为正常对照组，模型组，凉血解毒方高（Liangxue Jiedu groups with high，LXJD-H）、中（medium，LXJD-M）、低（low，LXJD-L）剂量组和甲氨蝶呤（methotrexate，MTX）组，观察小鼠背部皮损变化。采用银屑病皮损面积和疾病严重程度评分（psoriasis area and severity index，PASI）标准进行评分，光镜下观察皮肤组织形态学变化，测量表皮厚度;免疫荧光及组织化学法检测皮损中Ki67、CD3、CD11c的表达情况;流式细胞术检测脾脏中DCs数量;酶联免疫法（enzyme linked immunosorbent assay，ELISA）检测细胞上清液中细胞因子的量;采用RT-PCR法检测皮损组织中DCs相关受体基因水平;Western blotting法检测小鼠皮损中TLR7/TLR8通路相关蛋白表达。结果凉血解毒方明显降低皮损PASI评分及表皮厚度、减少炎性反应细胞的浸润;抑制皮损中TLR7通路相关蛋白的表达;体内外降低白细胞介素-23（interleukin-23，IL-23）、Toll样受体（Toll-like receptors，TLR7）、TLR8 及IL-12p40 mRNA表达及下调IL-23、IL-1β细胞因子分泌水平。结论凉血解毒方可以改善IMQ诱导的银屑病样小鼠皮损，且可以改善皮损中T淋巴细胞及DCs的浸润，同时抑制DCs的活化。

关键词: 银屑病, 咪喹莫特, 树突状细胞, 凉血解毒方

Abstract: Objective To observe the effects of Liangxue Jiedu decoction(LXJD) on the abnormal activation of dendritic cells on mice psoriasis-like lesions induced by imiquimod. Methods Thirty-six BALB/c mice were randomly divided into six groups:control group, model group, Liangxue Jiedu groups with high(LXJD-H),medium(LXJD-M) or low(LXJD-L) doses, and Methotrexate (MTX) group. The lesions were evaluated according to the psoriasis area and severity index(PASI). The histology and epidermal thicknesses were observed under light microscope.The expression of Ki-67 protein was detected by influorescent antibody technique. Meanwhile, the positive expression of CD3, CD11c was counted by immunohistochemical staining.The expression of dendritic cells in mouse spleen cells and the content of IL-23, IL-12p40, IL-1β in DC2.4 by flow cytometry. The IL-23 mRNA, TLR7, TLR8 were detected by PCR. The expression of TLR7 protein was detected by Western blotting. Results Compared with model group, the cutaneous symptoms in LXJD capsules groups were alleviated, with PASI scores decreased, epidermal parakeratosis and epidermal over-proliferation, the numbers of dermal T lymphocytes, dendritic cells reduced significantly.Compared with model group, the number of Ki67, CD3 and CD11c⁺ in LXJD group was reduced obviously. Meanwhile the expression of IL-23, TLR7, TLR8 and IL-12p40 were all decreased compared with model group. Then in the cell experiment the secretion of cytokines IL-23 and IL-1β were all decreased. Conclusion Liangxue Jiedu Decoction could improve IMQ-induced mouse psoriasis-like lesions and ameliorate the infiltration of T cells and DCs in psoriasis-like lesions, While inhibiting the activation of DCs.

Key words: psoriasis, imiquimod, dendritic cell, Liangxue Jiedu

中图分类号:

R289.5

王明星, 王燕, 赵京霞, 底婷婷, 阮智通, 蒙玉娇, 谢湘江, 张璐, 林燕, 王宁, 李萍. 树突状细胞在咪喹莫特诱导的银屑病样小鼠模型中的表达及凉血解毒方的干预作用[J]. 首都医科大学学报, 2017, 38(2): 249-259.

Wang Mingxing, Wang Yan, Zhao Jingxia, Di Tingting, Ruan Zhitong, Meng Yujiao, Xie Xiangjiang, Zhang Lu, Lin Yan, Wang Ning, Li Ping. Dendritic cells on mice psoriasis-like lesions induced by imiquimod and the effects of Liangxue Jiedu decoction[J]. Journal of Capital Medical University, 2017, 38(2): 249-259.

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树突状细胞在咪喹莫特诱导的银屑病样小鼠模型中的表达及凉血解毒方的干预作用

Dendritic cells on mice psoriasis-like lesions induced by imiquimod and the effects of Liangxue Jiedu decoction

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