Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155
10.13422/j.cnki.syfjx.20220779
- VernacularTitle:附子汤对类风湿性关节炎滑膜成纤维细胞MH7A增殖和miR-155表达的影响
- Author:
Wanli QIN
1
;
Yujie XU
1
;
Zhenzhen PAN
1
;
Xiaohui LI
1
;
Zhenhua WANG
1
;
Jianping SONG
1
;
Qin XU
1
;
Xinan HUANG
1
;
Changqing LI
1
Author Information
1. Guangzhou University of Chinese Medicine，Guangzhou 510405，China
- Publication Type:Journal Article
- Keywords:
Fuzitang;
rheumatoid arthritis;
MH7A cells;
miR-155;
SH2 domain-containing inositol 5-phosphatase-1（SHIP-1）;
phosphatidylinositol 3-kinase （PI3K）/protein kinase B 3 （Akt3）/mammalian target of rapamycin （mTOR） signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(14):29-35
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo observe the effects of Fuzitang （FZT） on the proliferation of MH7A cells， the human rheumatoid arthritis synovial fibroblasts， and the expression of miR-155 and explore its anti-rheumatoid arthritis mechanism. MethodMH7A cells were cultured in vitro and divided into a blank group， high- （25 g·L-1） and low-dose （12.5 g·L-1） FZT groups， and a positive drug group （hydroxychloroquine， 0.006 25 g·L-1）. The cell proliferation was detected by cell counting kit-8（CCK-8） method， and the change in the MH7A cell cycle was detected by flow cytometry. The mRNA expression of miR-155 and its downstream genes， including SH2 domain-containing inositol 5-phosphatase-1（SHIP-1）， protein kinase B 3（Akt3）， and mammalian target of rapamycin（mTOR）， was detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， and the protein expression of phosphatidylinositol 3-kinase （PI3K）， Akt3， and mTOR was detected by Western blot. ResultFZT in vitro in a concentration of 6.25 g·L-1 above could inhibit the proliferation of MH7A cells in the significant dose- and time-effect manner. Compared with the blank group， the FZT groups showed increased proportions of cells in the G2/M phase （P<0.05）， and the high-dose FZT group showed a decreased proportion of cells in the G0/G1 phase （P<0.05）. The arresting effect of FZT on the cell cycle was in a significant dose-effect manner. Compared with the blank group， the FZT groups showed down-regulated miR-155 and mTOR mRNA expression （P<0.05）， and the high-dose FZT group showed up-regulated SHIP1 mRNA expression and down-regulated Akt3 mRNA expression （P<0.05）. Compared with the blank group， the FZT groups showed reduced protein expression of PI3K， Akt3， and mTOR （P<0.05）. ConclusionFZT can significantly inhibit the proliferation of MH7A cells， and the mechanism is related to the promotion of the expression of SHIP-1 and down-regulation of the gene expression of the PI3K/Akt3/mTOR signaling pathway by down-regulating the expression of miR-155.
