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威灵仙总皂苷通过LncRNA OIP5-AS1/miR-410-3p/Wnt7b信号通路调控类风湿关节炎中滑膜成纤维样细胞增殖

Total Saponins of Radix Clematis Regulate Fibroblast-Like Synoviocyte Proliferation in Rheumatoid Arthritis via the LncRNA OIP5-AS1/MiR-410-3p/Wnt7b Signaling Pathway.

作者信息

Pan Lingyu, Sun Yuan, Jiang Hui, Chen Yan, Jiang Yeke, Han Yanquan, Wang Yongzhong

机构信息

The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, Anhui, China.

Anhui University of Chinese Medicine, Hefei 230031, Anhui, China.

出版信息

Evid Based Complement Alternat Med. 2022 May 27;2022:8393949. doi: 10.1155/2022/8393949. eCollection 2022.

DOI:10.1155/2022/8393949
PMID:35668775
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9166986/
Abstract

BACKGROUND

Rheumatoid arthritis (RA) is the most common autoimmune disease and affects multiple joints. Previous studies have shown that total saponins of (TSC) have a clear therapeutic effect on RA, but the specific mechanism has not yet been clarified. Literature screening and previous research suggest that the lncRNA OIP5-AS1/miR-410-3p/Wnt7b signaling pathway exerts a regulatory effect on the pathogenesis of RA. In this study, we examined whether the TSC treatment of RA affects the lncRNA OIP5-AS1/miR-410-3p/Wnt7b pathway.

MATERIALS AND METHODS

Freund's complete adjuvant was used to create an adjuvant arthritis (AA) rat model with rat synovial cells being harvested and cultured. The experiment comprises a normal group, model group, TSC optimal-dose group, TSC optimal-dose group + lncRNA OIP5-AS1siRNA group, lncRNA OIP5-AS1 siRNA group, and lncRNA OIP5-AS1 siRNA + NC group. MMT was used to screen the optimal concentration of TSC. The level of lncRNA OIP5-AS1, miR-410-3p, Wnt7b, -catenin, c-Myc, cyclin D1, GSK-3, and SFRP4 mRNA were detected by real-time-qPCR, the expression of Wnt7b, -catenin, c-Myc, cyclin D1, GSK-3, and p-GSK-3 (Ser9) protein were detected by immunofluorescence and Western blot.

RESULTS

We found that TSC inhibits the proliferation of RA FLS, TSC significantly reduced lncRNA OIP5-AS1, Wnt7b, -catenin, c-Myc, cyclin D1, and p-GSK-3/GSK-3 mRNA/protein expression, whereas the miR-410-3p and SFRP4 mRNA/protein expression levels were significantly upregulated. Our data suggest that TSC can inhibit the excessive proliferation of FLS to treat RA, the mechanism of which may be closely related to regulation of the lncRNA OIP5-AS1/miR-410-3p /Wnt7b signaling axis and the Wnt signaling pathway.

摘要

背景

类风湿关节炎（RA）是最常见的自身免疫性疾病，可累及多个关节。既往研究表明，[具体植物名称]总皂苷（TSC）对RA具有明确的治疗作用，但其具体机制尚未阐明。文献筛选及前期研究提示，lncRNA OIP5-AS1/miR-410-3p/Wnt7b信号通路对RA发病机制具有调控作用。本研究旨在探讨TSC治疗RA是否影响lncRNA OIP5-AS1/miR-410-3p/Wnt7b通路。

材料与方法

采用弗氏完全佐剂建立佐剂性关节炎（AA）大鼠模型，并收集大鼠滑膜细胞进行培养。实验分为正常组、模型组、TSC最佳剂量组、TSC最佳剂量组 + lncRNA OIP5-AS1 siRNA组、lncRNA OIP5-AS1 siRNA组、lncRNA OIP5-AS1 siRNA + NC组。采用MTT法筛选TSC的最佳浓度。通过实时定量PCR检测lncRNA OIP5-AS1、miR-410-3p、Wnt7b、β-连环蛋白、c-Myc、细胞周期蛋白D1、糖原合成酶激酶-3（GSK-3）和分泌型卷曲相关蛋白4（SFRP4）mRNA水平，通过免疫荧光和蛋白质印迹法检测Wnt7b蛋白、β-连环蛋白、c-Myc、细胞周期蛋白D1、GSK-3和磷酸化GSK-3（Ser9）的表达。

结果

我们发现TSC抑制RA成纤维样滑膜细胞（FLS）增殖，TSC显著降低lncRNA OIP5-AS1、Wnt7b、β-连环蛋白、c-Myc、细胞周期蛋白D1以及磷酸化GSK-3/GSK-3的mRNA/蛋白表达，而miR-410-3p和SFRP4的mRNA/蛋白表达水平显著上调。我们的数据表明，TSC可通过抑制FLS过度增殖来治疗RA，其机制可能与调控lncRNA OIP5-AS1/miR-410-3p/Wnt7b信号轴及Wnt信号通路密切相关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/c39a68ab62b8/ECAM2022-8393949.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/cc84448528d6/ECAM2022-8393949.001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/592df5bf1b28/ECAM2022-8393949.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/03b6ce4f3a65/ECAM2022-8393949.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/c39a68ab62b8/ECAM2022-8393949.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/cc84448528d6/ECAM2022-8393949.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/49c1718323d3/ECAM2022-8393949.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/f872909efceb/ECAM2022-8393949.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/6726eb7bcef8/ECAM2022-8393949.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/e7e781a847ec/ECAM2022-8393949.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/592df5bf1b28/ECAM2022-8393949.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/03b6ce4f3a65/ECAM2022-8393949.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ce3d/9166986/c39a68ab62b8/ECAM2022-8393949.008.jpg

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