Mechanism of paroxetine on osteoclast differentiation

doi:10.12307/2023.534

Abstract

Abstract: BACKGROUND: Excessive activation of osteoclasts is one of the key links in the occurrence and development of osteoporosis. Exploring the function of osteoclasts and developing drugs that can inhibit osteoclast differentiation and reduce bone resorption is an important means to explore the clinical treatment of osteoporosis.
OBJECTIVE: To analyze the mechanism of paroxetine on osteoclast differentiation based on the mitogen-activated protein kinase/nuclear factor-κB signal pathway.
METHODS: (1) In vitro experiment: Different concentrations of paroxetine were used to interfere with bone marrow-derived macrophages in mice, and then the effect of paroxetine on cell activity was determined by cell counting kit-8. Under the induction of macrophage colony stimulating factor and receptor activator of receptor activator of nuclear factor-κB ligand, different concentrations of paroxetine were used to interfere with osteoclast differentiation. The number of osteoclasts was determined by tartrate-resistant acid phosphatase staining. The effect of paroxetine on the mRNA expression of osteoclast differentiation related cytokines was detected by real-time fluorescence quantitative real-time PCR. The effect of paroxetine on nuclear factor κB and mitogen-activated protein kinase signal pathways in the cells was detected by western blot. (2) In vivo experiment: Forty male C57BL/6 mice were randomly divided into four groups (n=10 per group): blank control group, lipopolysaccharide group, paroxetine 2 mg/kg treatment group, and paroxetine 5 mg/kg treatment group. Lipopolysaccharide was injected subcutaneously into the sagittal suture of the skull every 2 days to construct the mouse skull osteolysis model in the lipopolysaccharide group, paroxetine 2 mg/kg and 5 mg/kg groups. The corresponding dose of paroxetine was injected 1 day after each lipopolysaccharide injection in the two paroxetine groups. After 14 days of treatment, mouse skulls were isolated for Micro-CT scanning.
RESULTS AND CONCLUSION: (1) In vitro experiment: there was no significant effect on the proliferation of bone marrow-derived macrophages when paroxetine concentration was ≤ 5 μmol/L, while paroxetine concentration > 10 μmol/L could inhibit cell proliferation. When paroxetine concentration was ≥ 0.5 μmol/L, there was a significant reduction in the number and morphology of osteoclast differentiation as well as the mRNA expression of c-Fos, Nfatc1, Ctsk, Mmp9, Acp5, and Atp6v0d2. Paroxetine at a dose of 0.5 μmol/L could inhibit the protein expression of c-Fos, Nfatc1, and Ctsk through activating nuclear factor-κB/mitogen-activated protein kinase signal pathway. (2) In vivo experiment: Micro-CT scanning results showed that compared with lipopolysaccharide, 5 mg/kg paroxetine could significantly increase the number of trabeculae and bone volume fraction and reduce bone surface bone volume ratio and trabecular separation (P < 0.05). (3) To conclude, paroxetine can inhibit bone resorption by reducing osteoclast differentiation and has a potential effect in the treatment of osteoporosis.

Key words: paroxetine, MAPK, nuclear factor κB, osteoclast, osteoporosis, bone marrow-derived macrophage

CLC Number:

Mo Yaomin, Liu Pan, Ma Ruixin, Zeng Gaofeng, Zong Shaohui. Mechanism of paroxetine on osteoclast differentiation[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(32): 5184-5190.

Figures/Tables 6

References

[1] BIJLSMA AY, MESKERS CG, WESTENDORP RG, et al. Chronology of age-related disease definitions: Osteoporosis and sarcopenia. Ageing Res Rev. 2012;11(2):320-324.
[2] ASPRAY TJ, HILL TR. Osteoporosis and the ageing skeleton. Subcell Biochem. 2019;91:453-476.
[3] HASEGAWA T, KIKUTA J, SUDO T, et al. Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by foxm1. Nat Immunol. 2019;20(12):1631-1643.
[4] BROWN JP. Long-term treatment of postmenopausal osteoporosis. Endocrinol Metab (Seoul). 2021;36(3):544-552.
[5] SHANE E, BURR D, ABRAHAMSEN B, et al. Atypical subtrochanteric and diaphyseal femoral fractures: Second report of a task force of the american society for bone and mineral research. J Bone Miner Res. 2014;29(1):1-23.
[6] LANGDAHL B. Treatment of postmenopausal osteoporosis with bone-forming and antiresorptive treatments: Combined and sequential approaches. Bone. 2020;139:115516.
[7] ZHAO X, PATIL S, XU F, et al. Role of biomolecules in osteoclasts and their therapeutic potential for osteoporosis. Biomolecules. 2021; 11(5):747.
[8] UDAGAWA N, KOIDE M, NAKAMURA M, et al. Osteoclast differentiation by rankl and opg signaling pathways. J Bone Miner Metab. 2021;39(1): 19-26.
[9] KIM JH, KIM K, KIM I, et al. Bcap promotes osteoclast differentiation through regulation of the p38-dependent creb signaling pathway. Bone. 2018;107:188-195.
[10] CHAWEEWANNAKORN W, ARIYOSHI W, OKINAGA T, et al. Ameloblastin attenuates rankl-mediated osteoclastogenesis by suppressing activation of nuclear factor of activated t-cell cytoplasmic 1 (nfatc1). J Cell Physiol. 2019;234(2):1745-1757.
[11] KOWALSKA M, NOWACZYK J, FIJALKOWSKI L, et al. Paroxetine-overview of the molecular mechanisms of action. Int J Mol Sci. 2021;22(4):1662.
[12] LIU RP, ZOU M, WANG JY, et al. Paroxetine ameliorates lipopolysaccharide-induced microglia activation via differential regulation of mapk signaling. J Neuroinflammation. 2014;11:47.
[13] ZHANG X, ZHU LB, HE JH, et al. Paroxetine suppresses reactive microglia-mediated but not lipopolysaccharide-induced inflammatory responses in primary astrocytes. J Neuroinflammation. 2020;17(1):50.
[14] XIE H, CHUNG DY, KURA S, et al. Differential effects of anesthetics on resting state functional connectivity in the mouse. J Cereb Blood Flow Metab. 2020;40(4):875-884.
[15] CARLSON EL, KARUPPAGOUNDER V, PINAMONT WJ, et al. Paroxetine-mediated grk2 inhibition is a disease-modifying treatment for osteoarthritis. Sci Transl Med. 2021;13(580):eaau8491.
[16] VERDELIS K, SALMON P. Microcomputed tomography imaging in odontogenesis studies. Methods Mol Biol. 2019;1922:309-324.
[17] JOHNSTON CB, DAGAR M. Osteoporosis in older adults. Med Clin North Am. 2020;104(5):873-884.
[18] KANIS JA, COOPER C, RIZZOLI R, et al. European guidance for the diagnosis and management of osteoporosis in postmenopausal women. Osteoporos Int. 2019;30(1):3-44.
[19] REID IR. A broader strategy for osteoporosis interventions. Nat Rev Endocrinol. 2020;16(6):333-339.
[20] FISCHER V, HAFFNER-LUNTZER M. Interaction between bone and immune cells: Implications for postmenopausal osteoporosis. Semin Cell Dev Biol. 2022;123:14-21.
[21] DA W, TAO L, ZHU Y. The role of osteoclast energy metabolism in the occurrence and development of osteoporosis. Front Endocrinol (Lausanne). 2021;12:675385.
[22] AMIN N, BOCCARDI V, TAGHIZADEH M, et al. Probiotics and bone disorders: The role of rankl/rank/opg pathway. Aging Clin Exp Res. 2020;32(3):363-371.
[23] HUANG L, CHEN W, WEI L, et al. Lonafarnib inhibits farnesyltransferase via suppressing erk signaling pathway to prevent osteoclastogenesis in titanium particle-induced osteolysis. Front Pharmacol. 2022;13:848152.
[24] XIAN Y, SU Y, LIANG J, et al. Oroxylin a reduces osteoclast formation and bone resorption via suppressing rankl-induced ros and nfatc1 activation. Biochem Pharmacol. 2021;193:114761.
[25] XU W, CHEN X, WANG Y, et al. Chitooligosaccharide inhibits rankl-induced osteoclastogenesis and ligation-induced periodontitis by suppressing mapk/ c-fos/nfatc1 signaling. J Cell Physiol. 2020;235(3): 3022-3032.
[26] CONG Q, JIA H, LI P, et al. P38alpha mapk regulates proliferation and differentiation of osteoclast progenitors and bone remodeling in an aging-dependent manner. Sci Rep. 2017;7:45964.
[27] LEE S, RAUCH J, KOLCH W. Targeting mapk signaling in cancer: Mechanisms of drug resistance and sensitivity. Int J Mol Sci. 2020; 21(3):1102.
[28] KANG JY, KANG N, YANG YM, et al. The role of ca(2+)-nfatc1 signaling and its modulation on osteoclastogenesis. Int J Mol Sci. 2020;21(10): 3646.
[29] YANG Y, CHUNG MR, ZHOU S, et al. Stat3 controls osteoclast differentiation and bone homeostasis by regulating nfatc1 transcription. J Biol Chem. 2019;294(42):15395-15407.
[30] XIAO L, ZHONG M, HUANG Y, et al. Puerarin alleviates osteoporosis in the ovariectomy-induced mice by suppressing osteoclastogenesis via inhibition of traf6/ros-dependent mapk/nf-kappab signaling pathways. Aging (Albany NY). 2020;12(21):21706-21729.
[31] KOGA Y, TSURUMAKI H, AOKI-SAITO H, et al. Roles of cyclic amp response element binding activation in the erk1/2 and p38 mapk signalling pathway in central nervous system, cardiovascular system, osteoclast differentiation and mucin and cytokine production. Int J Mol Sci. 2019;20(6):1346.
[32] YU H, LIN L, ZHANG Z, et al. Targeting nf-kappab pathway for the therapy of diseases: Mechanism and clinical study. Signal Transduct Target Ther. 2020;5(1):209.
[33] BARNABEI L, LAPLANTINE E, MBONGO W, et al. Nf-kappab: At the borders of autoimmunity and inflammation. Front Immunol. 2021; 12:716469.
[34] LAMPIASI N, RUSSO R, ZITO F. The alternative faces of macrophage generate osteoclasts. Biomed Res Int. 2016;2016:9089610.
[35] YASUDA H. Discovery of the rankl/rank/opg system. J Bone Miner Metab. 2021;39(1):2-11.
[36] SUL OJ, PARK HJ, SON HJ, et al. Lipopolysaccharide (lps)-induced autophagy is responsible for enhanced osteoclastogenesis. Mol Cells. 2017;40(11):880-887.
[37] CARR HS, CHANG JT, FROST JA. The pdz domain protein synj2bp regulates grk-dependent sst2a phosphorylation and downstream mapk signaling. Endocrinology. 2021;162(2):bqaa229.
[38] SUN X, ZHOU M, WEN G, et al. Paroxetine attenuates cardiac hypertrophy via blocking grk2 and adrb1 interaction in hypertension. J Am Heart Assoc. 2021;10(1):e016364.
[39] HU K, OLSEN BR. Osteoblast-derived vegf regulates osteoblast differentiation and bone formation during bone repair. J Clin Invest. 2016;126(2):509-526.
[40] CHEN X, WANG Z, DUAN N, et al. Osteoblast-osteoclast interactions. Connect Tissue Res. 2018;59(2):99-107.