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黄芩苷下调白细胞介素-1β刺激的鼻成纤维细胞外基质生成

Baicalin Down-Regulates IL-1β-Stimulated Extracellular Matrix Production in Nasal Fibroblasts.

作者信息

Shin Jae-Min, Kang Ju-Hyung, Lee Seoung-Ae, Park Il-Ho, Lee Heung-Man

机构信息

Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, South Korea.

Division of Brain Korea 21 Program for Biomedical Science, Korea University College of Medicine, Seoul, South Korea.

出版信息

PLoS One. 2016 Dec 21;11(12):e0168195. doi: 10.1371/journal.pone.0168195. eCollection 2016.

Abstract

PURPOSE

Baicalin, a Chinese herbal medicine, has anti-fibrotic and anti-inflammatory effects. The aims of present study were to investigate the effects of baicalin on the myofibroblast differentiation, extracellular matrix production, migration, and collagen contraction of interleukin (IL)-1β-stimulated nasal fibroblasts and to determine the molecular mechanism of baicalin in nasal fibroblasts.

METHODS

Nasal fibroblasts were isolated from the inferior turbinate of patients. Baicalin was used to treat IL-1β-stimulated nasal fibroblasts. To evaluate cytotoxicity, a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay was used. The expression levels of α-smooth muscle actin (SMA), fibronectin, phospho-mitogen-activated protein kinase (p-MAPK), p-Akt, p-p50, p-p65, and p-IκBα were measured by western blotting, reverse transcription-polymerase chain reaction (RT-PCR),or immunofluorescence staining. Fibroblast migration was analyzed with scratch assays and transwell migration assays. Total collagen was evaluated with the Sircol collagen assay. Contractile activity was measured with a collagen gel contraction assay.

RESULTS

Baicalin (0-50 μM) had no significant cytotoxic effects in nasal fibroblasts. The expression of α-SMA and fibronectin were significantly down-regulated in baicalin-treated nasal fibroblasts. Migration, collagen production, and contraction of IL-1β-stimulated nasal fibroblasts were significantly inhibited by baicalin treatment. Baicalin also significantly down-regulated p-MAPK, p-Akt, p-p50, p-p65, and p-IκBα in IL-1β-stimulated nasal fibroblasts.

CONCLUSIONS

We showed that baicalin down-regulated myofibroblast differentiation, extracellular matrix production, migration, and collagen contraction via the MAPK and Akt/ NF-κB pathways in IL-1β-stimulated nasal fibroblasts.

摘要

目的

黄芩苷是一种中药,具有抗纤维化和抗炎作用。本研究旨在探讨黄芩苷对白细胞介素(IL)-1β刺激的鼻成纤维细胞肌成纤维细胞分化、细胞外基质产生、迁移及胶原收缩的影响,并确定黄芩苷在鼻成纤维细胞中的分子机制。

方法

从患者下鼻甲分离出鼻成纤维细胞。用黄芩苷处理IL-1β刺激的鼻成纤维细胞。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基溴化四氮唑蓝法评估细胞毒性。通过蛋白质印迹法、逆转录-聚合酶链反应(RT-PCR)或免疫荧光染色法检测α-平滑肌肌动蛋白(SMA)、纤连蛋白、磷酸化丝裂原活化蛋白激酶(p-MAPK)、p-Akt、p-p50、p-p65和p-IκBα的表达水平。用划痕试验和Transwell迁移试验分析成纤维细胞迁移。用Sircol胶原分析法评估总胶原含量。用胶原凝胶收缩试验测量收缩活性。

结果

黄芩苷(0-50μM)对鼻成纤维细胞无明显细胞毒性作用。在黄芩苷处理的鼻成纤维细胞中,α-SMA和纤连蛋白的表达显著下调。黄芩苷处理显著抑制了IL-1β刺激的鼻成纤维细胞的迁移、胶原产生和收缩。黄芩苷还显著下调了IL-1β刺激的鼻成纤维细胞中的p-MAPK、p-Akt、p-p50、p-p65和p-IκBα。

结论

我们发现黄芩苷通过丝裂原活化蛋白激酶(MAPK)和Akt/核因子κB(NF-κB)信号通路下调IL-1β刺激的鼻成纤维细胞中的肌成纤维细胞分化、细胞外基质产生、迁移及胶原收缩。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f14f/5176301/9935d66a1de2/pone.0168195.g001.jpg

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