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

Received: 13 December 2021/Accepted: 11 March 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

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^*Corresponding author; email: qryangsdu@163.com