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水菖蒲含药血清对口腔黏膜成纤维细胞调控作用及复发性口腔溃疡治疗机制的研究

Investigation on Molecular Mechanism of Fibroblast Regulation and the Treatment of Recurrent Oral Ulcer by Shuizhongcao Granule-Containing Serum.

机构信息

Zhejiang Cancer Hospital, Hangzhou, Zhejiang 310022, China.

Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310053, China.

出版信息

Evid Based Complement Alternat Med. 2015;2015:324091. doi: 10.1155/2015/324091. Epub 2015 Oct 18.

Abstract

The purpose is to study the intervention, proliferation, and differentiation on fibroblast by Shuizhongcao Granule during the treatment of ROU and investigate the action mechanism in inflammatory microenvironment. Proliferation of rat fibroblasts was detected using CCK8. Western blot was used to detect the effect of drug-containing serum on the expression of protein associated with NF-κB and ERK pathway in rat fibroblasts. Expression of IL-10 and IL-12 was detected by PCR. Shuizhongcao Granule group successfully inhibited proliferation of rat fibroblast. Western blot results revealed that p65 and IKKB were significantly less expressed in Chinese medicine group, while pIκBα and pIKKαβ expression were significantly increased. We have also found that in this group the expression of pAKT was evidently suppressed and expression of pERK significantly decreased. PCR results showed significantly decreased expression levels of IL-10 and 1IL-12b in Chinese medicine group, while the expression of IL-12a was increased. Our results suggest that Shuizhongcao Granule can suppress the proliferation of fibroblast and inhibit the activation of NF-κB and thus suppress inflammatory reactions, possibly involving the inhibited expression of phosphorylated AKT, rather than the canonical pathway. Furthermore, it can inhibit ERK pathway and reduce IL-10 and IL-12b gene expression while enhancing IL-12a expression.

摘要

目的

研究水蟑螂颗粒在治疗复发性口疮(ROU)过程中对成纤维细胞的干预、增殖和分化作用,并探讨其在炎症微环境中的作用机制。采用 CCK8 检测大鼠成纤维细胞的增殖。Western blot 检测含药血清对大鼠成纤维细胞 NF-κB 和 ERK 通路相关蛋白表达的影响。采用 PCR 检测 IL-10 和 IL-12 的表达。水蟑螂颗粒组成功抑制了大鼠成纤维细胞的增殖。Western blot 结果显示,中药组 p65 和 IKKB 的表达明显减少,而 pIκBα 和 pIKKαβ 的表达明显增加。我们还发现,在该组中,pAKT 的表达明显受到抑制,pERK 的表达明显降低。PCR 结果显示,中药组 IL-10 和 1IL-12b 的表达水平明显降低,而 IL-12a 的表达增加。我们的结果表明,水蟑螂颗粒可抑制成纤维细胞的增殖,抑制 NF-κB 的激活,从而抑制炎症反应,可能涉及磷酸化 AKT 的表达抑制,而非经典途径。此外,它可以抑制 ERK 通路,降低 IL-10 和 IL-12b 基因表达,同时增强 IL-12a 表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a25e/4628681/1b8606ab5c5e/ECAM2015-324091.001.jpg

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