Sains Malaysiana 51(8)(2022): 2583-2593

http://doi.org/10.17576/jsm-2022-5108-18

IRE1α Promotes Cell Apoptosis and an Inflammatory Response in Endoplasmic Reticulum Stress-Induced Rheumatoid Arthritis Fibroblast-Like Synovial Cells by Enhancing Autophagy

(IRE1α Menggalakkan Apoptosis Sel dan Tindak Balas Keradangan dalam Retikulum Endoplasma yang Disebabkan oleh Tekanan Reumatoid Artritis Fibroblas Seperti Sel Sinovial dengan Meningkatkan Autofagi)

JIALIANG YANG^1,2, ZHENZHEN MA¹, QIAN JIA², YANSHAN LI², YUCHENG LU³ & QINGRUI YANG^1,*

¹Department of Rheumatology and Immunology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, 250021, China

²Department of Rheumatology and Immunology, Linyi People's Hospital, Linyi, Shandong, 276000, China

³Biobank, Linyi People's Hospital, Linyi, Shandong, 276000, China

Diserahkan: 13 Disember 2021/Diterima: 11 Mac 2022

Abstract

Endoplasmic reticulum (ER) stress can induce autophagy via the unfolded protein response (UPR), and autophagy can regulate the activation of inflammasomes. Inositol-requiring enzyme 1α (IRE1α) is a transducer of the UPR in cells with ER stress. Here, we investigated the role of IRE1α and its impact on ER stress in rheumatoid arthritis fibroblast-like synovial cells (RA-FLSs). RA-FLSs were isolated from rheumatoid arthritis (RA) patients and stimulated with thapsigargin (TG) to produce ER stress cells. ER stress-, autophagy and the expression of apoptosis-associated factors were investigated by western blotting and the qRT-PCR. Cellular ROS levels were assessed by flow cytometry. ELISAs were performed to determine the concentrations of inflammatory mediators. TG treatment promoted IRE1α, GRP78, CHOP, and ATP6 mRNA and protein expression. ROS generation was increased in TG-induced RA-FLSs; additionally, TG was found to induce cell inflammation by upregulating the expression of inflammasome markers and the concentrations of inflammatory mediators. The levels of autophagy markers, apoptosis-associated proteins, and mRNA were increased in TG-stimulated RA-FLSs. However, transfection with si-IRE1α suppressed TG-induced increases in ROS generation, inflammation levels, cell apoptosis, and autophagy in RA-FLSs. Treatment with the autophagy activator RAPA attenuated the protective effects of IRE1α silencing on TG-induced RA-FLS apoptosis and inflammatory damage. Our findings showed that in RA-FLSs, IRE1α silencing alleviated ER stress-induced inflammation and apoptosis caused by autophagy.

Keywords: Autophagy; endoplasmic reticulum stress; inositol-requiring enzyme 1α; rheumatoid arthritis fibroblast-like synovial cells; thapsigargin

Abstrak

Tekanan retikulum endoplasma (ER) boleh mendorong autofagi melalui tindak balas protein terungkap (UPR) dan autofagi boleh mengawal pengaktifan inflamasom. Enzim 1α yang memerlukan inositol (IRE1α) ialah transduser UPR dalam sel dengan tekanan ER. Di sini, kami mengkaji peranan IRE1α dan kesannya terhadap tekanan ER dalam sel sinovial seperti fibroblas artritis reumatoid (RA-FLSs). RA-FLS telah diasingkan daripada pesakit reumatoid artritis (RA) dan dirangsang dengan thapsigargin (TG) untuk menghasilkan sel tekanan ER. Tekanan ER, autofagi dan ekspresi faktor berkaitan apoptosis telah dikaji oleh pemedapan Western dan qRT-PCR. Tahap ROS sel dinilai oleh sitometri aliran. ELISA dilakukan untuk menentukan kepekatan mediator keradangan. Rawatan TG menggalakkan ekspresi mRNA dan protein IRE1α, GRP78, CHOP dan ATP6. Penjanaan ROS telah meningkat dalam RA-FLS yang disebabkan oleh TG; tambahan TG didapati mendorong keradangan sel dengan mengawal selia ekspresi penanda inflamasom dan kepekatan mediator keradangan. Tahap penanda autofagi, protein berkaitan apoptosis dan mRNA telah meningkat dalam RA-FLS yang dirangsang TG. Walau bagaimanapun, pemindahan dengan si-IRE1α menindas peningkatan yang disebabkan oleh TG dalam penjanaan ROS, tahap keradangan, apoptosis sel dan autofagi dalam RA-FLSs. Rawatan dengan pengaktif autofagi RAPA melemahkan kesan perlindungan pembungkaman IRE1α pada apoptosis RA-FLS yang disebabkan oleh TG dan kerosakan keradangan. Penemuan kami menunjukkan bahawa dalam RA-FLSs, pembungkaman IRE1α mengurangkan keradangan dan apoptosis yang disebabkan oleh tekanan ER yang disebabkan oleh autofagi.

Kata kunci: Autofagi; enzim 1α yang memerlukan inositol; reumatoid artritis fibroblas seperti sel sinovial; tekanan retikulum endoplasma; thapsigargin

RUJUKAN

Ahmadiany, M., Alavi-Samani, M., Hashemi, Z., Moosavi, M.A. & Rahmati, M. 2019. The increased RNAse activity of ire1α in pbmcs from patients with rheumatoid arthritis. Advanced Pharmaceutical Bulletin 9(3): 505-509.

Aletaha, D., Neogi, T., Silman, A.J., Funovits, J., Felson, D.T., Bingham III, C.O., Birnbaum, N.S., Burmester, G.R., Bykerk, V.P., Cohen, M.D., Combe, B., Costenbader, K.H., Dougados, M., Emery, R., Ferraccioli, G., Johanna, M.W., Hobbs, K., Huizinga, T.W.J., Kavanaugh, A., Kay, J., Kvien, T.K., Laing, T., Mease, P., Ménard, H.A., Moreland, L.W., Naden, R.L., Pincus, T., Smolen, J.S., Stanislawska-Biernat, E., Symmons, D., Tak, P.P., Upchurch, K.S., Vencovský, J., Wolfe, F. & Hawker, G. 2010. Rheumatoid arthritis classification criteria: An American College of Rheumatology/European League against rheumatism collaborative initiative. Annals of the Rheumatic Diseases 69(9): 1580-1588.

Asif, A.M., Fox, D.A. & Ruth, J.H. 2017. Synovial cellular and molecular markers in rheumatoid arthritis. Seminars in Immunopathology 39(4): 385-393.

Bronner, D.N., Abuaita, B.H., Chen, X., Fitzgerald, K.A., Nunez, G., He, Y., Yin, X. & O'Riordan, M.X.D. 2015. Endoplasmic reticulum stress activates the inflammasome via NLRP3- and Caspase-2-driven mitochondrial damage. Immunity 43(3): 451-462.

Chen, H.G., Han, H.Z., Li, Y., Yu, Y.H. & Xie, K.L. 2020. Hydrogen alleviated organ injury and dysfunction in sepsis: The role of cross-talk between autophagy and endoplasmic reticulum stress. Experimental Research International Immunopharmacology 78: 106049.

Chong, W.C., Shastri, M.D. & Eri, R. 2017. Endoplasmic reticulum stress and oxidative stress: A vicious nexus implicated in bowel disease pathophysiology. International Journal of Molecular Sciences 18(4): 771.

De Cock, D. & Hyrich, K. 2018. Malignancy and rheumatoid arthritis: Epidemiology, risk factors and management. Best Practice & Research Clinical Rheumatology 32(6): 869-886.

Gao, G., Chen, W., Yan, M., Liu, J., Luo, H., Wang, C. & Yang, P. 2020. Rapamycin regulates the balance between cardiomyocyte apoptosis and autophagy in chronic heart failure by inhibiting mTOR signaling. International Journal of Molecular Medicine 45: 195-209.

Gao, Y., Zhu, H., Yang, F., Wang, Q., Feng, Y. & Zhang, C. 2018 Glucocorticoid-activated IRE1α/XBP-1s signaling: An autophagy-associated protective pathway against endotheliocyte damage. American Journal of Physiology Cell Physiology 315: C300-C309.

Han, J. & Kaufman, R.J. 2017. Physiological/pathological ramifications of transcription factors in the unfolded protein response. Genes & Development 31(14): 1417-1438.

Hetz, C. 2012. The unfolded protein response: Controlling cell fate decisions under ER stress and beyond. Nature Reviews Molecular Cell Biology 13: 89-102.

Hong, J., Kim, K., Park, E., Lee, J., Markofski, M.M., Marrelli, S.P. & Park, Y. 2018. Exercise ameliorates endoplasmic reticulum stress-mediated vascular dysfunction in mesenteric arteries in atherosclerosis. Scientific Reports 8: 7938.

Hou, Y., Fu, J., Sun, S., Jin, Y., Wang, X. & Zhang, L. 2019. BDE-209 induces autophagy and apoptosis via IRE1α/Akt/mTOR signaling pathway in human umbilical vein endothelial cells. Environmental Pollution 253: 429-438.

Hu, H., Wang, C., Jin, Y., Meng, Q., Liu, Q., Liu, Z., Liu, K., Liu, X. & Sun, H. 2019. Catalpol inhibits homocysteine-induced oxidation and inflammation via inhibiting Nox4/NF-κB and GRP78/PERK pathways in human aorta. Endothelial Cells Inflammation 42: 64-80.

Kabala, P.A., Angiolilli, C., Yeremenko, N., Grabiec, A.M., Giovannone, B., Pots, D., Radstake, T.R., Baeten, D. & Reedquist, K.A. 2017. Endoplasmic reticulum stress cooperates with Toll-like receptor ligation in driving activation of rheumatoid arthritis fibroblast-like synoviocytes. Arthritis Research & Therapy 19: 207.

Kato, M., Ospelt, C., Gay, R.E, Gay, S. & Klein, K. 2014. Dual role of autophagy in stress-induced cell death in rheumatoid arthritis synovial fibroblasts. Arthritis & Rheumatology 66(1): 40-48.

Kong, F.J., Ma, L.L., Guo, J.J., Xu, L.H., Li, Y. & Qu, S. 2018. Endoplasmic reticulum stress/autophagy pathway is involved in diabetes-induced neuronal apoptosis and cognitive decline in mice. Clinical Science 132(1): 111-125.

Liu, T. 2019. Regulation of inflammasome by autophagy. Advances in Experimental Medicine and Biology 1209: 109-123.

Maurel, M., Chevet, E., Tavernier, J. & Gerlo, S. 2014. Getting RIDD of RNA: IRE1 in cell fate regulation. Trends in Biochemical Sciences 39(5): 245-254.

Moore, K.A. & Hollien, J. 2012. The unfolded protein response in secretory cell function. Annual Review of Genetics 46: 165-183.

Navid, F. & Colbert, R.A. 2017. Causes and consequences of endoplasmic reticulum stress in rheumatic disease. Nature Reviews Rheumatology 13: 25-40.

Pirmardvand, C.S., Varshosaz, J. & Taymouri, S. 2018. Recent approaches for targeted drug delivery in rheumatoid arthritis diagnosis and treatment. Artificial Cells, Nanomedicine, and Biotechnology 46: 502-514.

Qi, L., Tsai, B. & Arvan, P. 2017. New insights into the physiological role of endoplasmic reticulum-associated degradation. Trends in Cell Biology 27(6): 430-440.

Qiu, Q., Zheng, Z., Chang, L., Zhao, Y.S., Tan, C., Dandekar, A., Zhang, Z., Lin, Z., Gui, M., Li, X., Zhang, T., Kong, Q., Li, H., Chen, S., Chen, A., Kaufman, R.J., Yang, W.L., Lin, H.K., Zhang, D., Perlman, H., Thorp, E., Zhang, K. & Fang, D. 2013. Toll-like receptor-mediated IRE1α activation as a therapeutic target for inflammatory arthritis. The EMBO Journal 32: 2477-2490.

Rahmati, M., Moosavi, M.A. & McDermott, M.F. 2018. ER stress: A therapeutic target in rheumatoid arthritis? Trends in Pharmacological Sciences 39(7): 610-623.

Rockel, J.S. & Kapoor, M. 2017 Autophagy: Controlling cell fate in rheumatic diseases. Nature Reviews Rheumatology 13(3): 193.

Shin, Y.J., Han, S.H., Kim, D.S., Lee, G.H., Yoo, W.H., Kang, Y.M., Choi, J.Y., Lee, Y.C., Park, S.J., Jeong, S.K., Kim, H.T., Chae, S.W., Jeong, H.J., Kim, H.R. & Chae, H.J. 2010. Autophagy induction and CHOP under-expression promotes survival of fibroblasts from rheumatoid arthritis patients under endoplasmic reticulum stress. Arthritis Research & Therapy 12: R19.

Song, S., Tan, J., Miao, Y., Li, M. & Zhang, Q. 2017. Crosstalk of autophagy and apoptosis: Involvement of the dual role of autophagy under ER stress. Journal of Cellular Physiology 232(11): 2977-2984.

Song, S., Tan, J., Miao, Y. & Zhang, Q. 2018. Crosstalk of ER stress-mediated autophagy and ER-phagy: Involvement of UPR and the core autophagy machinery. Journal of Cellular Physiology 233(5): 3867-3874.

Song, Y., Rivera, C., Mai, J. & Sun, A.W.L. 2020. Splicing HAC1/XBP1 mRNAs in cytoplasm: The non-conventional mRNA splicing reaction in the unfolded protein response OBM. Genetics 4(2): 11.

Sparks, J.A. 2019. Rheumatoid arthritis. Annals of Internal Medicine 170(1): ITC1-ITC16.

Tang, C.H. 2020 Research of pathogenesis and novel therapeutics in arthritis 2.0. International Journal of Molecular Sciences 21(21): 8125.

Vomero, M., Barbati, C., Colasanti, T., Perricone, C., Novelli, L., Ceccarelli, F., Spinelli, F.R., Franco, M.D., Conti, F., Valesini, G. & Alessandro, C. 2018. Autophagy and rheumatoid arthritis: Current knowledges and future perspectives. Frontiers in Immunology 9: 1577.

Wang, L., Dong, H., Song, G., Zhang, R., Pan, J. & Han, J. 2018. TXNDC5 synergizes with HSC70 to exacerbate the inflammatory phenotype of synovial fibroblasts in rheumatoid arthritis through NF-κB signaling. Cellular & Molecular Immunology 15(7): 685-696.

Wu, P., Tian, T., Zhao, J., Song, Q., Wu, X., Guo, Y., Yu, Y., Tan, S. & Xia, H. 2020. IRE1α-JNK pathway-mediated autophagy promotes cell survival in response to endoplasmic reticulum stress during the initial phase of hepatic steatosis. Life Sciences 264: 118668.

Wu, Y., Wang, X., Guo, H., Zhang, B., Zhang, X.B., Shi, Z.J. & Yu, L. 2013. Synthesis and screening of 3-MA derivatives for autophagy inhibitors. Autophagy 9(4): 595-603.

Yamasaki, S., Yagishita, N., Tsuchimochi, K., Kato, Y., Ssaki, T., Amano, T., Beppu, M., Aoki, H., Nakamura, H., Nishioka, K. & Nakajima, T. 2006. Resistance to endoplasmic reticulum stress is an acquired cellular characteristic of rheumatoid synovial cells. International Journal of Molecular Medicine 18(1): 113-117.

Zhang, X., Hu, P., Ding, S.Y., Sun, T., Liu, L., Han, S., DeLeo, A.B., Sadagopan, A., Guo, W. & Wang, H. 2019. Induction of autophagy-dependent apoptosis in cancer cells through activation of ER stress: An uncovered anti-cancer mechanism by anti-alcoholism drug disulfiram. American Journal of Cancer Research 9(6): 1266-1281.

Zhou, W., Shen, Q., Wang, H., Yang, J., Zhangm, C., Deng, Z., Wu, K., Zhou, Y., Zheng, J., Zhang, Y. & Shen, W. 2020. Knockdown of YAP/TAZ inhibits the migration and invasion of fibroblast synovial cells in rheumatoid arthritis by regulating autophagy. Journal of Immunology Research 2020: 9510594.

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