Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155

Wanli QIN; Yujie XU; Zhenzhen PAN; Xiaohui LI; Zhenhua WANG; Jianping SONG; Qin XU; Xinan HUANG; Changqing LI

doi:10.13422/j.cnki.syfjx.20220779

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Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155

更新时间：2022-09-01

- Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 14, Pages: 29-35(2022)
- 作者机构：
  
  广州中医药大学，广州 510405
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20220779
  CLC： R2-0;R22;R285.5;R289;R33
- Published：20 July 2022，
  
  Published Online：30 January 2022，
  
  Received：09 November 2021，
- 稿件说明：
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覃万莉,徐玉洁,潘真真等.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[J].中国实验方剂学杂志,2022,28(14):29-35.

QIN Wanli,XU Yujie,PAN Zhenzhen,et al.Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(14):29-35.
覃万莉,徐玉洁,潘真真等.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[J].中国实验方剂学杂志,2022,28(14):29-35. DOI： 10.13422/j.cnki.syfjx.20220779.

QIN Wanli,XU Yujie,PAN Zhenzhen,et al.Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(14):29-35. DOI： 10.13422/j.cnki.syfjx.20220779.

摘要

目的

观察附子汤对类风湿性关节炎滑膜成纤维细胞MH7A增殖的影响，研究附子汤对miR-155表达的影响并进一步探讨其抗类风湿性关节炎的作用机制。

方法

体外培养MH7A细胞，设空白组、附子汤高剂量组（25 g·L

-1

）、低剂量组（12.5 g·L

-1

）和硫酸羟氯喹阳性药组（0.006 25 g·L

-1

），采用细胞增殖与活性检测（CCK-8）试剂盒法检测细胞增殖，流式细胞术检测MH7A细胞周期的改变；应用实时荧光定量聚合酶链式反应（Real-time PCR）法检测药物处理后miR-155及其下游基因含SH2结构域的肌醇5-磷酸酶-1（SHIP-1）、蛋白激酶B3（Akt3）和哺乳动物雷帕霉素靶蛋白（mTOR） mRNA表达，蛋白免疫印迹法（Western blot）检测磷脂酰肌醇3-激酶（PI3K）、Akt3和mTOR的蛋白表达。

结果

附子汤体外在质量浓度6.25 g·L

-1

以上时，能明显抑制MH7A增殖，且呈明显的量-效关系和时-效关系。与空白组比较，附子汤高、低剂量组G

/M期细胞比例均明显增加（

0.05），高剂量组G

期细胞比例明显降低（

0.05），其细胞周期的阻滞作用呈明显的量效关系。与空白组比较，附子汤高、低剂量组miR-155的表达均明显下调（

0.05）；附子汤高剂量组SHIP1 mRNA表达明显上调、Akt3 mRNA表达明显下调（

0.05），附子汤高、低剂量组mTOR mRNA表达均明显下调（

0.05）。与空白组比较，附子汤高、低剂量组的PI3K、Akt3和mTOR的蛋白表达均明显下调（

0.05）。

结论

附子汤对MH7A细胞增殖具有显著的抑制作用，作用机制与下调miR-155的表达，从而促进SHIP-1的表达，抑制其下游PI3K/Akt3/mTOR信号通路的基因表达有关。

Abstract

Objective

To 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.

Method

MH7A 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.

Result

FZT

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 G

/M phase （

0.05）， and the high-dose FZT group showed a decreased proportion of cells in the G

phase （

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 （

0.05）， and the high-dose FZT group showed up-regulated SHIP1 mRNA expression and down-regulated Akt3 mRNA expression （

0.05）. Compared with the blank group， the FZT groups showed reduced protein expression of PI3K， Akt3， and mTOR （

0.05）.

Conclusion

FZT 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.

关键词

附子汤类风湿性关节炎MH7A细胞miR-155肌醇5-磷酸酶-1（SHIP-1）磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B3（Akt3）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路

Keywords

Fuzitangrheumatoid arthritisMH7A cellsmiR-155SH2 domain-containing inositol 5-phosphatase-1（SHIP-1）phosphatidylinositol 3-kinase （PI3K）/protein kinase B 3 （Akt3）/mammalian target of rapamycin （mTOR） signaling pathway

references

PICERNO V，FERRO F，ADINOLFI A，et al.One year in review： The pathogenesis of rheumatoid arthritis［J］.Clin Exp Rheumatol，2015，33（4）：551-558.

LEE S Y，CHO M L，OH H J，et al.Interleukin-2/anti-interleukin-2 monoclonal antibody immune complex suppresses collagen-induced arthritis in mice by fortifying interleukin-2/STAT5 signalling pathways［J］.Immunology，2012，137（4）：305-316.

COOLES F A，ISAACS J D，ANDERSON A E.Treg cells in rheumatoid arthritis： An update［J］.Curr Rheumatol Rep，2013，15（9）：352.

KUROWSKA-STOLARSKA M，ALIVERNINI S，BALLANTINE L E，et al.MicroRNA-155 as a proinflammatory regulator in clinical and experimental arthritis［J］.Proc Natl Acad Sci U S A，2011，108（27）：11193-11198.

李赛美，李宇航.“十二五”普通高等教育本科国家级规划教材.伤寒论讲义［M］.3版.北京：人民卫生出版社，2017：208-209.

MIYAZAWA K，MORI A，OKUDAIRA H.Establishment and characterization of a novel human rheumatoid fibroblast-like synoviocyte line， MH7A， immortalized with SV40 T antigen［J］.J Biochem，1998，124（6）：1153-1162.

HUBER L C，DISTLER O，TARNER I，et al.Synovial fibroblasts： Key players in rheumatoid arthritis［J］.Rheumatology （Oxford），2006，45（6）：669-675.

MEINECKE I，RUTKAUSKAITE E，GAY S，et al.The role of synovial fibroblasts in mediating joint destruction in rheumatoid arthritis［J］.Curr Pharm Des，2005，11（5）：563-568.

SMOLEN J S，ALETAHA D，KOELLER M，et al.New therapies for treatment of rheumatoid arthritis［J］.Lancet，2007，370（9602）：1861-1874.

MCINNES I B，SCHETT G.The pathogenesis of rheumatoid arthritis［J］.N Engl J Med，2011，365（23）：2205-2219.

FIRESTEIN G S.Invasive fibroblast-like synoviocytes in rheumatoid arthritis. Passive responders or transformed aggressors？［J］.Arthritis Rheum，1996，39（11）：1781-1790.

FOLKMAN J.Angiogenesis in cancer， vascular， rheumatoid and other disease［J］.Nat Med，1995，1（1）：27-31.

KOCH A E，HARLOW L A，HAINES G K，et al.Vascular endothelial growth factor. A cytokine modulating endothelial function in rheumatoid arthritis［J］.J Immunol，1994，152（8）：4149-4156.

MANN M，MEHTA A，ZHAO J L，et al.An NF-κB-microRNA regulatory network tunes macrophage inflammatory responses［J］.Nat Commun，2017，8（1）：851.

SHARMA A R，SHARMA G，LEE S S，et al.miRNA-regulated key components of cytokine signaling pathways and inflammation in rheumatoid arthritis［J］.Med Res Rev，2016，36（3）：425-439.

WANG J，ZHAO Q.LncRNA LINC-PINT increases SOCS1 expression by sponging miR-155-5p to inhibit the activation of ERK signaling pathway in rheumatoid arthritis synovial fibroblasts induced by TNF-α［J］.Int Immunopharmacol，2020，84：106497.

BLÜMLS，BONELLIM，NIEDERREITERB，et al.Essential role of microRNA-155 in the pathogenesis of autoimmune arthritis in mice［J］.Arthritis Rheum，2011，63（5）：1281-1288.

SU L C，HUANG A F，JIA H，et al.Role of microRNA-155 in rheumatoid arthritis［J］.Int J Rheum Dis，2017，20（11）：1631-1637.

NAKANO N，NISHIYAMA C，YAGITA H，et al.Notch signaling enhances FcεRI-mediated cytokine production by mast cells through direct and indirect mechanisms［J］.J Immunol，2015，194（9）：4535-4544.

XIONG Y，MEDVEDEV A E.Induction of endotoxin tolerance in vivo inhibits activation of IRAK4 and increases negative regulators IRAK-M， SHIP-1， and A20［J］.J Leukoc Biol，2011，90（6）：1141-1148.

张妍，陆芸.磷酸酶SHIP-1对T细胞分化、稳态和免疫应答的调控效应［J］.复旦学报：医学版，2014，41（3）：412-415，426.

SMITH M D，WEEDON H，PAPANGELIS V，et al.Apoptosis in the rheumatoid arthritis synovial membrane： Modulation by disease-modifying anti-rheumatic drug treatment［J］.Rheumatology （Oxford），2010，49（5）：862-875.

QU Y，WU J，DENG J X，et al.MicroRNA-126 affects rheumatoid arthritis synovial fibroblast proliferation and apoptosis by targeting PIK3R2 and regulating PI3K-Akt signal pathway［J］.Oncotarget，2016，7（45）：74217-74226.

KIM J，JUNG K H，YOO J，et al.PBT-6， a novel PI3KC2γ inhibitor in rheumatoid arthritis［J］.Biomol Ther （Seoul），2020，28（2）：172-183.

KIM K W，CHO M L，PARK M K，et al.Increased interleukin-17 production via a phosphoinositide 3-kinase/Akt and nuclear factor kappaB-dependent pathway in patients with rheumatoid arthritis［J］.Arthritis Res Ther，2005，7（1）：R139-R148.

TSAI C H，LIU S C，WANG Y H，et al.Osteopontin inhibition of miR-129-3p enhances IL-17 expression and monocyte migration in rheumatoid arthritis［J］.Biochim Biophys Acta Gen Subj，2017，1861（2）：15-22.

DINESH P，RASOOL M.Berberine inhibits IL-21/IL-21R mediated inflammatory proliferation of fibroblast-like synoviocytes through the attenuation of PI3K/Akt signaling pathway and ameliorates IL-21 mediated osteoclastogenesis［J］.Cytokine，2018，106：54-66.

MITRA A，RAYCHAUDHURI S K，RAYCHAUDHURI S P.IL-22 induced cell proliferation is regulated by PI3K/Akt/mTOR signaling cascade［J］.Cytokine，2012，60（1）：38-42.

WU L，GUO Q，YANG J，et al.Tumor necrosis factor alpha promotes osteoclast formation via PI3K/Akt pathway-mediated blimp1 expression upregulation［J］.J Cell Biochem，2017，118（6）：1308-1315.

CHOE J Y，HUN KIM J ，PARK K Y，et al.Activation of dickkopf-1 and focal adhesion kinase pathway by tumour necrosis factor α induces enhanced migration of fibroblast-like synoviocytes in rheumatoid arthritis［J］.Rheumatology （Oxford），2016，55（5）：928-938.

LIACINI A，SYLVESTER J，LI W Q，et al.Induction of matrix metalloproteinase-13 gene expression by TNF-alpha is mediated by MAP kinases， AP-1， and NF-kappaB transcription factors in articular chondrocytes［J］.Exp Cell Res，2003，288（1）：208-217.

XUAN W，FENG X，QIAN C，et al.Osteoclast differentiation gene expression profiling reveals chemokine CCL4 mediates RANKL-induced osteoclast migration and invasion via PI3K pathway［J］.Cell Biochem Funct，2017，35（3）：171-177.

WANG C H，CHEN L N，ZHU P，et al.CD147 stimulates the angiogenesis in rheumatoid synovium via the activation of vascular endothelial growth factor［J］.Chin J Cell Mol Immunol，2007，23（5）：426-428.

ZOU L，ZHANG G，LIU L，et al.Relationship between PI3K pathway and angiogenesis in CIA rat synovium［J］.Am J Transl Res，2016，8（7）：3141-3147.

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