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苦参方提取物的治疗作用及其对接触性皮炎模型小鼠嗜酸性粒细胞趋化因子的抑制作用。

Therapeutic action of Kushen recipe extractive and its inhibitory effect on eotaxin in mouse models with contact dermatitis.

机构信息

Department of Health Management, Beidaihe Rehabilitation and Rest Center of PLA, Qinhuangdao, China.

Department of Pharmacy, Shanghai Skin Disease Hospital, Tongji University, Shanghai, China.

出版信息

Allergol Immunopathol (Madr). 2023 Jul 1;51(4):110-123. doi: 10.15586/aei.v51i4.793. eCollection 2023.

Abstract

BACKGROUND

Treatment of skin allergic diseases remains a challenging research topic.

OBJECTIVE

To investigate the effect of Kushen recipe extractive (KS) gel on contact dermatitis (CD) of mouse.

METHODS

Allergic contact dermatitis (ACD) model of mouse was established. Immunohistochemical method (ICH) and flow cytometry method (FCM) were used to detect CD4 and CD8 T lymphocytes and explore the regulation effect of KS on the immune status of the organism. The expression status of eotaxin tissue was evaluated by real-time polymerase chain reaction (RT-PCR), ICH, and western blotting method. The survival rates of HaCaT cell and Fibroblasts affected by KS were detected by methyl thiazolyl tetrazolium (MTT) method. The inhibitory effect of KS on eotaxin produced by HaCaT cell and FBs induced by TNF-α and interleukin (IL)-4 were evaluated using RT-PCR and enzyme-linked immunosorbent assay methods. The inhibitory effect of KS on nuclear factor-κB (NF-κB) and Signal transducers and activators of transcription 6 (STAT6) activation induced by TNF-α and IL-4 was detected by electrophoretic mobility shift assay and western blotting methods.

RESULTS

We confirmed that KS shows favorable therapeutic effect on CD, which can obviously inhibit eotaxin expression and Eosinophils recruitment in allergic skin of mouse, as well as regulate the immune status of the organism. Furthermore, KS and its main effective components can inhibit TNF-α and IL-4 induced upregulation of eotaxin via the two signal transduction pathways, NF-κB and STAT6.

CONCLUSIONS

The great importance of traditional Chinese recipe KS is evidenced by its therapeutic effect and mechanism in ACD of mouse.

摘要

背景

皮肤过敏性疾病的治疗仍然是一个具有挑战性的研究课题。

目的

研究苦参配方提取物(KS)凝胶对小鼠接触性皮炎(CD)的作用。

方法

建立小鼠过敏性接触性皮炎(ACD)模型。采用免疫组织化学(ICH)和流式细胞术(FCM)检测 CD4 和 CD8 T 淋巴细胞,探讨 KS 对机体免疫状态的调节作用。采用实时聚合酶链反应(RT-PCR)、ICH 和蛋白质印迹法检测组织嗜酸性粒细胞趋化因子(eotaxin)的表达状态。采用噻唑蓝(MTT)法检测 KS 对 HaCaT 细胞和受 KS 影响的成纤维细胞存活率的影响。采用 RT-PCR 和酶联免疫吸附试验(ELISA)方法评价 KS 对 TNF-α和白细胞介素(IL)-4诱导的 HaCaT 细胞和 FBs 产生的 eotaxin 的抑制作用。采用电泳迁移率变动分析和蛋白质印迹法检测 KS 对 TNF-α和 IL-4 诱导的核因子-κB(NF-κB)和信号转导和转录激活因子 6(STAT6)激活的抑制作用。

结果

我们证实 KS 对 CD 具有良好的治疗作用,能明显抑制小鼠过敏性皮肤中 eotaxin 的表达和嗜酸性粒细胞的募集,并能调节机体的免疫状态。此外,KS 及其主要有效成分可通过两条信号转导通路 NF-κB 和 STAT6 抑制 TNF-α和 IL-4 诱导的 eotaxin 上调。

结论

传统中药 KS 在治疗小鼠 ACD 中的疗效和作用机制证明了其重要性。

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