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
发布时间：2019-05-07 02:11
【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~(-1)2p40 mRNA表达及下调IL-23、IL~(-1)β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。
[Abstract]:Aim to observe the intervention effect of Liangxue jiedu recipe on activated dendritic cells (dendritic cells,DCs) induced by imiquimod (imiquimod,IMQ) in silver chip-like mice. Methods the mice were randomly divided into normal control group, model group, high (Liangxue Jiedu groups with high,LXJD-H group, middle dose group (medium,LXJD-M), low dose group (low,LXJD-L) and methotrexate (methotrexate,MTX) group. The changes of the skin lesions in the back of mice were observed. The psoriasis lesion area and disease severity score (psoriasis area and severity index,PASI) was used to evaluate the histological changes of the skin and the thickness of the skin under light microscope. Immunofluorescence and histochemistry were used to detect the expression of Ki67,CD3,CD11c, flow cytometry was used to detect the number of DCs in spleen, enzyme-linked immunosorbent assay (enzyme linked immunosorbent assay,ELISA) was used to detect the amount of cytokines in the supernatant. RT-PCR method was used to detect the level of DCs-associated receptor gene in the lesions.; Western blotting method was used to detect the expression of TLR7/TLR8 pathway-related proteins in the lesions of mice. Results Liangxuejiedu recipe significantly decreased the PASI score and epidermal thickness, decreased the infiltration of inflammatory response cells, and inhibited the expression of TLR7 pathway-related proteins in the lesions. It decreased the expression of interleukin-23,IL-23), Toll-like receptor (Toll-like receptors,TLR7), TLR8 and IL~ (- 1) 2p40 mRNA and down-regulated the secretion of IL-23,IL~ (- 1) 尾 cytokines in vitro and in vivo. Conclusion Liangxuejiedu recipe can improve the IMQ-induced lesions of psoriasis-like mice, improve the infiltration of T lymphocytes and DCs in the lesions, and inhibit the activation of DCs at the same time.
【作者单位】： 首都医科大学附属北京中医医院;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;北京中医药大学;
【基金】：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)~~
【分类号】：R285.5;R-332
本文编号：2470684
[Abstract]:Aim to observe the intervention effect of Liangxue jiedu recipe on activated dendritic cells (dendritic cells,DCs) induced by imiquimod (imiquimod,IMQ) in silver chip-like mice. Methods the mice were randomly divided into normal control group, model group, high (Liangxue Jiedu groups with high,LXJD-H group, middle dose group (medium,LXJD-M), low dose group (low,LXJD-L) and methotrexate (methotrexate,MTX) group. The changes of the skin lesions in the back of mice were observed. The psoriasis lesion area and disease severity score (psoriasis area and severity index,PASI) was used to evaluate the histological changes of the skin and the thickness of the skin under light microscope. Immunofluorescence and histochemistry were used to detect the expression of Ki67,CD3,CD11c, flow cytometry was used to detect the number of DCs in spleen, enzyme-linked immunosorbent assay (enzyme linked immunosorbent assay,ELISA) was used to detect the amount of cytokines in the supernatant. RT-PCR method was used to detect the level of DCs-associated receptor gene in the lesions.; Western blotting method was used to detect the expression of TLR7/TLR8 pathway-related proteins in the lesions of mice. Results Liangxuejiedu recipe significantly decreased the PASI score and epidermal thickness, decreased the infiltration of inflammatory response cells, and inhibited the expression of TLR7 pathway-related proteins in the lesions. It decreased the expression of interleukin-23,IL-23), Toll-like receptor (Toll-like receptors,TLR7), TLR8 and IL~ (- 1) 2p40 mRNA and down-regulated the secretion of IL-23,IL~ (- 1) 尾 cytokines in vitro and in vivo. Conclusion Liangxuejiedu recipe can improve the IMQ-induced lesions of psoriasis-like mice, improve the infiltration of T lymphocytes and DCs in the lesions, and inhibit the activation of DCs at the same time.
【作者单位】： 首都医科大学附属北京中医医院;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;北京中医药大学;
【基金】：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)~~
【分类号】：R285.5;R-332
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