Abstract
In our previous study, we explored the therapeutic effect of berberine (BBR) against IL-21/IL-21R mediated inflammatory proliferation of adjuvant-induced arthritic fibroblast-like synoviocytes (AA-FLS) through the PI3K/Akt pathway. The current study was designed to explore the therapeutic potential of BBR (15–45 µM) against IL-21/IL-21R mediated autophagy in AA-FLS mediated through PI3K/Akt signaling and Th17/Treg imbalance. Upon IL-21 stimulation, AA-FLS expressed elevated levels of autophagy-related 5 (Atg5), Beclin-1 and LC3-phosphatidylethanolamine conjugate 3-II (LC3-II) through the utilization of p62 and inhibition of C/EBP homologous protein (CHOP). BBR (15–45 µM) inhibited autophagy in AA-FLS cells mediated through PI3K/Akt signaling via suppressing autophagic elements, p62 sequestration and induction of CHOP in a dose-dependent manner. Moreover, IL-21 promoted the uncontrolled proliferation of AA-FLS through induction of B cell lymphoma-2 (Bcl-2) and diminished expression of Bcl-2 associated X protein (BAX) via PI3K/Akt signaling. BBR inhibited the proliferation of AA-FLS via promoting apoptosis through increased expression of BAX and diminished Bcl-2 transcription factor levels. Furthermore, T cells stimulated with IL-21 induced CD4+ CD196+ Th17 cells proliferation through RORγt activation mediated in a PI3K/Akt dependent manner. BBR inhibited the proliferation of Th17 cells through downregulation of RORγt in a concentration-dependent manner. BBR also promoted the differentiation of CD4+ CD25+ Treg cells through induction of forkhead box P3 (Foxp3) activation via aryl hydrocarbon receptor (AhR) and upregulation of cytochrome P450 family 1, subfamily A, polypeptide 1 (CYP1A1). Collectively, we conclude that BBR might attenuate AA-FLS proliferation through inhibition of IL-21/IL-21R dependent autophagy and regulates the Th17/Treg imbalance in RA.
Similar content being viewed by others
References
Bordy R, Totoson P, Prati C et al (2018) Microvascular endothelial dysfunction in rheumatoid arthritis. Nat Rev Rheumatol 14:404–420. https://doi.org/10.1038/s41584-018-0022-8
Weyand CM, Goronzy JJ (2017) Immunometabolism in early and late stages of rheumatoid arthritis. Nat Rev Rheumatol 13:291–301. https://doi.org/10.1038/nrrheum.2017.49
Tu J, Hong W, Zhang P et al (2018) Ontology and function of fibroblast-like and macrophage-like synoviocytes: how do they talk to each other and can they Be targeted for rheumatoid arthritis therapy? Front Immunol 9:1467–1477. https://doi.org/10.3389/fimmu.2018.01467
Buckley CD, McGettrick HM (2018) Leukocyte trafficking between stromal compartments: lessons from rheumatoid arthritis. Nat Rev Rheumatol 14:476–487. https://doi.org/10.1038/s41584-018-0042-4
Shikhagaie MM, Germar K, Bal SM et al (2017) Innate lymphoid cells in autoimmunity: emerging regulators in rheumatic diseases. Nat Rev Rheumatol 13:164–173. https://doi.org/10.1038/nrrheum.2016.218
Chen Z, Bozec A, Ramming A, Schett G (2019) Anti-inflammatory and immune-regulatory cytokines in rheumatoid arthritis. Nat Rev Rheumatol 15:9–17. https://doi.org/10.1038/s41584-018-0109-2
Falconer J, Murphy AN, Young SP et al (2018) Synovial cell metabolism and chronic inflammation in rheumatoid arthritis. Arthritis Rheumatol (Hoboken, NJ) 70:984–999. https://doi.org/10.1002/art.40504
Dinesh P, Rasool M (2018) 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. Cytokine 106:54–66. https://doi.org/10.1016/j.cyto.2018.03.005
Varricchi G, Harker J, Borriello F et al (2016) T follicular helper (Tfh) cells in normal immune responses and in allergic disorders. Allergy 71:1086–1094. https://doi.org/10.1111/all.12878
Diehl SA, Schmidlin H, Nagasawa M et al (2012) IL-6 triggers IL-21 production by human CD4+ T cells to drive STAT3-dependent plasma cell differentiation in B cells. Immunol Cell Biol 90:802–811. https://doi.org/10.1038/icb.2012.17
Rao DA, Gurish MF, Marshall JL et al (2017) Pathologically expanded peripheral T helper cell subset drives B cells in rheumatoid arthritis. Nature 542:110–114
Niu X, He D, Zhang X et al (2010) IL-21 regulates Th17 cells in rheumatoid arthritis. Hum Immunol 71:334–341. https://doi.org/10.1016/j.humimm.2010.01.010
Sakuraba K, Oyamada A, Fujimura K et al (2016) Interleukin-21 signaling in B cells, but not in T cells, is indispensable for the development of collagen-induced arthritis in mice. Arthritis Res Ther 18:1–10. https://doi.org/10.1186/s13075-016-1086-y
Xing R, Jin Y, Sun L et al (2016) Interleukin-21 induces migration and invasion of fibroblast-like synoviocytes from patients with rheumatoid arthritis. Clin Exp Immunol 184:147–158. https://doi.org/10.1111/cei.12751
Roeleveld DM, Marijnissen RJ, Walgreen B et al (2017) Higher efficacy of anti-IL-6/IL-21 combination therapy compared to monotherapy in the induction phase of Th17-driven experimental arthritis. PLoS ONE 12:e0171757–e0171773. https://doi.org/10.1371/journal.pone.0171757
Sanchez-Martin P, Komatsu M (2018) p62/SQSTM1—steering the cell through health and disease. J Cell Sci 131:21–34. https://doi.org/10.1242/jcs.222836
Ye X, Zhou XJ, Zhang H (2018) Exploring the role of autophagy-related gene 5 (ATG5) yields important insights into autophagy in autoimmune/autoinflammatory diseases. Front Immunol 9:2334–2349. https://doi.org/10.3389/fimmu.2018.02334
Vomero M, Barbati C, Colasanti T et al (2018) Autophagy and rheumatoid arthritis: current knowledges and future perspectives. Front Immunol 9:1577–1587. https://doi.org/10.3389/fimmu.2018.01577
Chen YM, Chang CY, Chen HH et al (2018) Association between autophagy and inflammation in patients with rheumatoid arthritis receiving biologic therapy. Arthritis Res Ther 20:268–279. https://doi.org/10.1186/s13075-018-1763-0
Shin YJ, Han SH, Kim DS et al (2010) Autophagy induction and CHOP under-expression promotes survival of fibroblasts from rheumatoid arthritis patients under endoplasmic reticulum stress. Arthritis Res Ther 12:R19–R30. https://doi.org/10.1186/ar2921
Ireland JM, Unanue ER (2011) Autophagy in antigen-presenting cells results in presentation of citrullinated peptides to CD4 T cells. J Exp Med 208:2625–2632. https://doi.org/10.1084/jem.20110640
Xu K, Xu P, Yao JF et al (2013) Reduced apoptosis correlates with enhanced autophagy in synovial tissues of rheumatoid arthritis. Inflamm Res 62:229–237. https://doi.org/10.1007/s00011-012-0572-1
Kato M, Ospelt C, Gay RE et al (2014) Dual role of autophagy in stress-induced cell death in rheumatoid arthritis synovial fibroblasts. Arthritis Rheumatol (Hoboken, NJ) 66:40–48. https://doi.org/10.1002/art.38190
Meng Q, Du X, Wang H et al (2017) Astragalus polysaccharides inhibits cell growth and pro-inflammatory response in IL-1beta-stimulated fibroblast-like synoviocytes by enhancement of autophagy via PI3K/AKT/mTOR inhibition. Apoptosis 22:1138–1146. https://doi.org/10.1007/s10495-017-1387-x
Connor AM, Mahomed N, Gandhi R et al (2012) TNFalpha modulates protein degradation pathways in rheumatoid arthritis synovial fibroblasts. Arthritis Res Ther 14:62–71. https://doi.org/10.1186/ar3778
Kim EK, Kwon JE, Lee SY et al (2017) IL-17-mediated mitochondrial dysfunction impairs apoptosis in rheumatoid arthritis synovial fibroblasts through activation of autophagy. Cell Death Dis 8:e2565–e2575. https://doi.org/10.1038/cddis.2016.490
Chang L, Feng X, Gao W (2018) Proliferation of rheumatoid arthritis fibroblast-like synoviocytes is enhanced by IL-17-mediated autophagy through STAT3 activation. Connect Tissue Res. https://doi.org/10.1080/03008207.2018.1552266
Feng FB, Qiu HY (2018) Effects of Artesunate on chondrocyte proliferation, apoptosis and autophagy through the PI3K/AKT/mTOR signaling pathway in rat models with rheumatoid arthritis. Biomed Pharmacother 102:1209–1220. https://doi.org/10.1016/j.biopha.2018.03.142
Hernandez-Palma LA, Garcia-Arellano S, Bucala R et al (2018) Functional MIF promoter haplotypes modulate Th17-related cytokine expression in peripheral blood mononuclear cells from control subjects and rheumatoid arthritis patients. Cytokine 115:89–96. https://doi.org/10.1016/j.cyto.2018.11.014
Marijnissen RJ, Roeleveld DM, Young D et al (2014) Interleukin-21 receptor deficiency increases the initial toll-like receptor 2 response but protects against joint pathology by reducing Th1 and Th17 cells during streptococcal cell wall arthritis. Arthritis Rheumatol 66:886–895. https://doi.org/10.1002/art.38312
Pfeifle R, Rothe T, Ipseiz N et al (2017) Regulation of autoantibody activity by the IL-23-TH17 axis determines the onset of autoimmune disease. Nat Immunol 18:104–113. https://doi.org/10.1038/ni.3579
Korn T, Bettelli E, Oukka M, Kuchroo VK (2009) IL-17 and Th17 Cells. Annu Rev Immunol 27:485–517. https://doi.org/10.1146/annurev.immunol.021908.132710
Rasmussen TK, Andersen T, Hvid M et al (2010) Increased interleukin 21 (IL-21) and IL-23 are associated with increased disease activity and with radiographic status in patients with early rheumatoid arthritis. J Rheumatol 37:2014–2020. https://doi.org/10.3899/jrheum.100259
Kondo Y, Yokosawa M, Kaneko S et al (2018) Review: transcriptional regulation of CD4+ T cell differentiation in experimentally induced arthritis and rheumatoid arthritis. Arthritis Rheumatol (Hoboken, NJ) 70:653–661. https://doi.org/10.1002/art.40398
Tong B, Yuan X, Dou Y et al (2016) Norisoboldine, an isoquinoline alkaloid, acts as an aryl hydrocarbon receptor ligand to induce intestinal Treg cells and thereby attenuate arthritis. Int J Biochem Cell Biol 75:63–73. https://doi.org/10.1016/j.biocel.2016.03.014
Nikiphorou E, Buch MH, Hyrich KL (2017) Biologics registers in RA: methodological aspects, current role and future applications. Nat Rev Rheumatol 13:503–510. https://doi.org/10.1038/nrrheum.2017.81
Zhang X, Zhao Y, Xu J et al (2015) Modulation of gut microbiota by berberine and metformin during the treatment of high-fat diet-induced obesity in rats. Sci Rep 5:14405–14415. https://doi.org/10.1038/srep14405
Pozsgay J, Szekanecz Z, Sarmay G (2017) Antigen-specific immunotherapies in rheumatic diseases. Nat Rev Rheumatol 13:525–537. https://doi.org/10.1038/nrrheum.2017.107
Wang Z, Chen Z, Yang S et al (2014) Berberine ameliorates collagen-induced arthritis in rats associated with anti-inflammatory and anti-angiogenic effects. Inflammation 37:1789–1798. https://doi.org/10.1007/s10753-014-9909-y
Wang X, He X, Zhang CF et al (2017) Anti-arthritic effect of berberine on adjuvant-induced rheumatoid arthritis in rats. Biomed Pharmacother 89:887–893. https://doi.org/10.1016/j.biopha.2017.02.099
Deng Y, Xu J, Zhang X et al (2014) Berberine attenuates autophagy in adipocytes by targeting BECN1. Autophagy 10:1776–1786. https://doi.org/10.4161/auto.29746
Tong B, Yuan X, Dou Y et al (2016) Sinomenine induces the generation of intestinal Treg cells and attenuates arthritis via activation of aryl hydrocarbon receptor. Lab Invest 96:1076–1086. https://doi.org/10.1038/labinvest.2016.86
Yue M, Xia Y, Shi C et al (2017) Berberine ameliorates collagen-induced arthritis in rats by suppressing Th17 cell responses via inducing cortistatin in the gut. FEBS J 284:2786–2801. https://doi.org/10.1111/febs.14147
Li R, Cai L, Tang W et al (2016) Apoptotic effect of geniposide on fibroblast-like synoviocytes in rats with adjuvant-induced arthritis via inhibiting ERK signal pathway in vitro. Inflammation 39:30–38. https://doi.org/10.1007/s10753-015-0219-9
Crowley T, O’Neil JD, Adams H et al (2017) Priming in response to pro-inflammatory cytokines is a feature of adult synovial but not dermal fibroblasts. Arthritis Res Ther 19:35–46. https://doi.org/10.1186/s13075-017-1248-6
McInnes IB, Schett G (2017) Pathogenetic insights from the treatment of rheumatoid arthritis. Lancet (London, England) 389:2328–2337. https://doi.org/10.1016/S0140-6736(17)31472-1
Burmester GR, Pope JE (2017) Novel treatment strategies in rheumatoid arthritis. Lancet (London, England) 389:2338–2348. https://doi.org/10.1016/S0140-6736(17)31491-5
Yuan MJ, Wang T (2016) Advances of the interleukin-21 signaling pathway in immunity and angiogenesis. Biomed reports 5:3–6. https://doi.org/10.3892/br.2016.665
Arbogast F, Gros F (2018) Lymphocyte autophagy in homeostasis, activation, and inflammatory diseases. Front Immunol 9:1801–1819. https://doi.org/10.3389/fimmu.2018.01801
Lin NY, Beyer C, Giessl A et al (2013) Autophagy regulates TNFalpha-mediated joint destruction in experimental arthritis. Ann Rheum Dis 72:761–768. https://doi.org/10.1136/annrheumdis-2012-201671
Yang Z, Fujii H, Mohan SV et al (2013) Phosphofructokinase deficiency impairs ATP generation, autophagy, and redox balance in rheumatoid arthritis T cells. J Exp Med 210:2119–2134. https://doi.org/10.1084/jem.20130252
Yin H, Wu H, Chen Y et al (2018) The therapeutic and pathogenic role of autophagy in autoimmune diseases. Front Immunol 9:1512–1523. https://doi.org/10.3389/fimmu.2018.01512
DeSelm CJ, Miller BC, Zou W et al (2011) Autophagy proteins regulate the secretory component of osteoclastic bone resorption. Dev Cell 21:966–974. https://doi.org/10.1016/j.devcel.2011.08.016
Xu K, Cai Y, Lu S-M et al (2015) Autophagy induction contributes to the resistance to methotrexate treatment in rheumatoid arthritis fibroblast-like synovial cells through high mobility group box chromosomal protein 1. Arthritis Res Ther 17:374–384. https://doi.org/10.1186/s13075-015-0892-y
Li S, Chen JW, Xie X et al (2017) Autophagy inhibitor regulates apoptosis and proliferation of synovial fibroblasts through the inhibition of PI3K/AKT pathway in collagen-induced arthritis rat model. Am J Transl Res 9:2065–2076
Yang R, Zhang Y, Wang L et al (2017) Increased autophagy in fibroblast-like synoviocytes leads to immune enhancement potential in rheumatoid arthritis. Oncotarget 8:15420–15430. https://doi.org/10.18632/oncotarget.14331
Wang X, Jiang S, Sun Q (2011) Effects of berberine on human rheumatoid arthritis fibroblast-like synoviocytes. Exp Biol Med (Maywood) 236:859–866. https://doi.org/10.1258/ebm.2011.010366
Wang X, Yang C, Xu F et al (2018) Imbalance of circulating Tfr/Tfh ratio in patients with rheumatoid arthritis. Clin Exp Med 19:55–64. https://doi.org/10.1007/s10238-018-0530-5
Zhao C, Gu Y, Zeng X, Wang J (2018) NLRP3 inflammasome regulates Th17 differentiation in rheumatoid arthritis. Clin Immunol 197:154–160. https://doi.org/10.1016/j.clim.2018.09.007
Kaneko S, Kondo Y, Yokosawa M et al (2018) The RORgammat-CCR60-CCL20 axis augments Th17 cells invasion into the synovia of rheumatoid arthritis patients. Mod Rheumatol 28:814–825. https://doi.org/10.1080/14397595.2017.1416923
Chen W, Wang J, Xu Z et al (2018) Apremilast ameliorates experimental arthritis via suppression of Th1 and Th17 cells and enhancement of CD4(+)Foxp3(+) regulatory T cells differentiation. Front Immunol 9:1662–1672. https://doi.org/10.3389/fimmu.2018.01662
Azizi G, Jadidi-Niaragh F, Mirshafiey A (2013) Th17 cells in immunopathogenesis and treatment of rheumatoid arthritis. Int J Rheum Dis 16:243–253. https://doi.org/10.1111/1756-185X.12132
Cui H, Cai Y, Wang L et al (2018) Berberine regulates Treg/Th17 balance to treat ulcerative colitis through modulating the gut microbiota in the colon. Front Pharmacol 9:571–588. https://doi.org/10.3389/fphar.2018.00571
Acknowledgments
Palani Dinesh would like to thank Council of Scientific and Industrial Research (CSIR) for providing financial assistance in the form of Senior Research Fellowship (SRF), [Acknowledgment no: 112290/2K17/1; File no: 09/844(0059)/2018].
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that there is no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Dinesh, P., Rasool, M. Berberine mitigates IL-21/IL-21R mediated autophagic influx in fibroblast-like synoviocytes and regulates Th17/Treg imbalance in rheumatoid arthritis. Apoptosis 24, 644–661 (2019). https://doi.org/10.1007/s10495-019-01548-6
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10495-019-01548-6