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维生素 D 结合蛋白和 1,25(OH)2D3 对类风湿关节炎滑膜成纤维细胞活力和凋亡的相加作用。

Additive Effects of VDBP and 1,25(OH)2D3 on the Viability and Apoptosis of Rheumatoid Arthritis Synovial Fibroblasts.

机构信息

Department of Orthopedic Surgery, Shandong Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.

Shandong Provincial Key Laboratory for Rheumatic Disease and Translational Medicine, Shandong Qianfoshan Hospital, Cheeloo College of Medicine, Shandong University, Jinan, China.

出版信息

Front Endocrinol (Lausanne). 2021 Jan 28;11:583229. doi: 10.3389/fendo.2020.583229. eCollection 2020.

DOI:10.3389/fendo.2020.583229
PMID:33584536
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7876401/
Abstract

AIM

This study is to investigate the additive effect of Vitamin D-binding protein (VDBP) and 1,25(OH)2D3 on the viability and apoptosis of synovial cells from patients with rheumatoid arthritis (RA).

METHODS

Synovial tissues and synovial fluid of patients with RA and osteoarthritis (OA) were collected. The expression of VDBP was analyzed with immunohistochemistry and ELISA. CCK-8 assay was applied to detect cell viability. Flow cytometry was used to analyze cell cycle and apoptosis.

RESULTS

Immunohistochemical results showed that the expression of VDBP in the synovium of RA patients was significantly lower than that of OA (P<0.05). Similarly, ELISA results presented a lower expression of VDBP in the synovial fluid of RA patients. The results of CCK-8 assay showed that both 1,25(OH)2D3 and VDBP significantly inhibited the viability of rheumatoid arthritis synovial fibroblasts (RASF) (P<0.05). The treatment with 1,25(OH)2D3+VDBP led to more significantly inhibited viability of RASF, compared with 1,25(OH)2D3 alone (P<0.05). The results of flow cytometry showed that 1,25(OH)2D3 and VDBP both promoted the apoptosis of RASF (P<0.05) and 1,25(OH)2D3+VDBP led to a higher proportion of RASF apoptosis, compared with 1,25(OH)2D3 alone (P<0.05). However, 1,25(OH)2D3 and VDBP had no significant effect on the cell cycle of RASF. Additionally, 1,25(OH)2D3 promoted the expression of VDBP in RASF, but not concentration-dependently.

CONCLUSION

VDBP is reduced in the synovial tissue and synovial fluid of RA patients and can inhibit viability of RASF and promote the apoptosis of RASF. The 1,25(OH)2D3 can upregulate the expression of VDBP in RASF. Additionally, VDBP can enhance the effects of 1,25(OH)2D3 on viability and apoptosis of RASF.

摘要

目的

本研究旨在探讨维生素 D 结合蛋白 (VDBP) 和 1,25(OH)2D3 对类风湿关节炎 (RA) 患者滑膜细胞活力和凋亡的相加作用。

方法

收集 RA 和骨关节炎 (OA) 患者的滑膜组织和滑液，采用免疫组化和 ELISA 分析 VDBP 的表达。CCK-8 检测细胞活力，流式细胞术分析细胞周期和凋亡。

结果

免疫组化结果显示，RA 患者滑膜中 VDBP 的表达明显低于 OA（P<0.05）。同样，ELISA 结果显示 RA 患者滑液中 VDBP 的表达较低。CCK-8 检测结果表明，1,25(OH)2D3 和 VDBP 均显著抑制类风湿关节炎滑膜成纤维细胞 (RASF) 的活力（P<0.05）。与单独使用 1,25(OH)2D3 相比，联合使用 1,25(OH)2D3+VDBP 导致 RASF 活力的抑制更为显著（P<0.05）。流式细胞术结果显示，1,25(OH)2D3 和 VDBP 均促进 RASF 凋亡（P<0.05），与单独使用 1,25(OH)2D3 相比，联合使用 1,25(OH)2D3+VDBP 导致 RASF 凋亡比例更高（P<0.05）。然而，1,25(OH)2D3 和 VDBP 对 RASF 细胞周期没有显著影响。此外，1,25(OH)2D3 促进 RASF 中 VDBP 的表达，但非浓度依赖性。

结论

RA 患者滑膜组织和滑液中 VDBP 减少，可抑制 RASF 活力并促进 RASF 凋亡。1,25(OH)2D3 可上调 RASF 中 VDBP 的表达。此外，VDBP 可增强 1,25(OH)2D3 对 RASF 活力和凋亡的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/fb78b98f255b/fendo-11-583229-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/c4a90efb388d/fendo-11-583229-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/68e927eb95cf/fendo-11-583229-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/bd2d0509152d/fendo-11-583229-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/fa510899ae89/fendo-11-583229-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/fb78b98f255b/fendo-11-583229-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/c4a90efb388d/fendo-11-583229-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/68e927eb95cf/fendo-11-583229-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/bd2d0509152d/fendo-11-583229-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/fa510899ae89/fendo-11-583229-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b8a/7876401/fb78b98f255b/fendo-11-583229-g005.jpg

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