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新型胸腺免疫抑制五肽TIPP对RBL-2H3细胞IgE介导的激活作用的抑制

Inhibition of the IgE-mediated activation of RBL-2H3 cells by TIPP, a novel thymic immunosuppressive pentapeptide.

作者信息

Lian Qianqian, Cheng Yanna, Zhong Chuanqing, Wang Fengshan

机构信息

Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.

Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China.

出版信息

Int J Mol Sci. 2015 Jan 20;16(1):2252-68. doi: 10.3390/ijms16012252.

DOI:10.3390/ijms16012252
PMID:25608657
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4307361/
Abstract

TIPP is a novel thymic immunosuppressive pentapeptide originally obtained from calf thymic immunosuppressive extract. The present study aimed to investigate the inhibitory activity of TIPP on IgE-mediated activation of RBL-2H3 cells. Release of β-hexosaminidase and histamine, intracellular calcium, membrane ruffling, mRNA levels of cytokines, cyclooxygenase-2 (COX-2) expression, and activation of mitogen-activated protein kinases (MAP kinases) and NF-κB were determined by colorimetric assay, fluorescence spectrophotometer, confocal fluorescence microscope, quantification PCR, and Western blot, respectively. The results showed that TIPP significantly inhibited the degranulation in IgE-antigen complex-stimulated RBL-2H3 cells without cytotoxicity. TIPP significantly suppressed the increase of intracellular calcium and the rearrangement of F-actin, attenuated the transcription of pro-inflammatory cytokines (IL-3, -4, -6, -13, TNF-α, and monocyte chemotactic protein-1 (MCP-1)), and decreased the expression of COX-2. Western blot analysis showed that TIPP had an inhibitory activity on the phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and ERK kinase 1/2 (MEK1/2), and inhibited the activation of NF-κB. The data suggested that TIPP effectively suppressed IgE-mediated activation of RBL-2H3 cells via blocking MEK/ERK and NF-κB signaling pathways.

摘要

TIPP是一种新型胸腺免疫抑制五肽，最初从牛胸腺免疫抑制提取物中获得。本研究旨在探讨TIPP对IgE介导的RBL-2H3细胞激活的抑制活性。分别通过比色法、荧光分光光度计、共聚焦荧光显微镜、定量PCR和蛋白质免疫印迹法测定β-己糖胺酶和组胺的释放、细胞内钙、膜皱襞、细胞因子的mRNA水平、环氧化酶-2（COX-2）的表达以及丝裂原活化蛋白激酶（MAP激酶）和NF-κB的激活。结果表明，TIPP显著抑制IgE-抗原复合物刺激的RBL-2H3细胞脱颗粒，且无细胞毒性。TIPP显著抑制细胞内钙的增加和F-肌动蛋白的重排，减弱促炎细胞因子（IL-3、-4、-6、-13、TNF-α和单核细胞趋化蛋白-1（MCP-1））的转录，并降低COX-2的表达。蛋白质免疫印迹分析表明，TIPP对细胞外信号调节蛋白激酶1/2（ERK1/2）和ERK激酶1/2（MEK1/2）的磷酸化具有抑制活性，并抑制NF-κB的激活。数据表明，TIPP通过阻断MEK/ERK和NF-κB信号通路有效抑制IgE介导的RBL-2H3细胞激活。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/1f57bc55dcd9/ijms-16-02252-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/510fca2693ed/ijms-16-02252-g001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/9f742ea01ac8/ijms-16-02252-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/06a9367400d1/ijms-16-02252-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/a3c427130c9c/ijms-16-02252-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/1f57bc55dcd9/ijms-16-02252-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/510fca2693ed/ijms-16-02252-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/dba9c42d3a17/ijms-16-02252-g002a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/c62b8fe0b91b/ijms-16-02252-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/e60300a139b2/ijms-16-02252-g004a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/3ed007080a55/ijms-16-02252-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/9f742ea01ac8/ijms-16-02252-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/06a9367400d1/ijms-16-02252-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/a3c427130c9c/ijms-16-02252-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f35/4307361/1f57bc55dcd9/ijms-16-02252-g009.jpg

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