조골세포 소포체 스트레스로 인한 골 손실을 개선하기 위한 운동효과 물질이 포함된 extracellular vehicles의 효과
Effect of Extracellular Vehicles, Including Exercise Mimetics, for Improving Bone Loss Caused by Osteoblast Endoplasmic Reticulum Stress
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
PURPOSE: This study aims to investigate the bone-muscle crosstalk mechanism by treating dysfunctional osteoblasts with extracellular vehicles (EVs) containing exercise mimetics released during muscle contraction, with the intention of identifying key factors in regulatory mechanisms. METHODS: To understand the bone-muscle crosstalk mechanism, the effects of treatment were evaluated in osteoblasts and C2C12. First, palmitate was administered to osteoblasts to induce bone loss due to endoplasmic reticulum (ER) stress. For validation, two groups were evaluated: a control group (CON) and the palmitate treatment group (PAL). Then, C2C12 myotubes were stimulated with electrical pulses to induce muscle contraction, and the released extracellular vehicles (EVs) were harvested and applied to osteoblasts. The results were used to determine the relevant mechanisms for improving bone loss. The impact of EVs released from muscles on osteoblasts was examined using the CD63-GFP plasmid. Four groups were used to compare across the different variables of interest: CON; EV, EV-treated group; PAL; EP, EV+palmitate-treated group. RESULTS: The palmitate-treated osteoblasts showed increased expression of inositol-requiring protein 1α (IRE1α), decreased sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA), reduced ER Ca2+, and mitochondrial membrane potential (MMP), indicating induced ER stress. Palmitate-induced ER stress was associated with elevated nuclear factor kappa B (NFκB) and receptor activator of NFκB (RANKL) levels and an increased RANKL/osteoprotegerin (OPG) ratio, causing bone loss. The treatment of osteoblasts with EVs from muscle contraction was found to improve ER stress, the levels of ER Ca2+, MMP, NFκB, and RANKL, and the RANKL/OPG ratio. CONCLUSIONS: Palmitate reduces ER Ca2+ and induces osteoblast ER stress while impairing mitochondrial function. Furthermore, through the RANKL/RANK/OPG pathway, a key mechanism related to bone loss is induced. However, substances induced by muscle contraction can act on osteoblasts through exosomes, potentially improving this process.
Abstract
PURPOSE: This study aims to investigate the bone-muscle crosstalk mechanism by treating dysfunctional osteoblasts with extracellular vehicles (EVs) containing exercise mimetics released during muscle contraction, with the intention of identifying key factors in regulatory mechanisms. METHODS: To understand the bone-muscle crosstalk mechanism, the effects of treatment were evaluated in osteoblasts and C2C12. First, palmitate was administered to osteoblasts to induce bone loss due to endoplasmic reticulum (ER) stress. For validation, two groups were evaluated: a control group (CON) and the palmitate treatment group (PAL). Then, C2C12 myotubes were stimulated with electrical pulses to induce muscle contraction, and the released extracellular vehicles (EVs) were harvested and applied to osteoblasts. The results were used to determine the relevant mechanisms for improving bone loss. The impact of EVs released from muscles on osteoblasts was examined using the CD63-GFP plasmid. Four groups were used to compare across the different variables of interest: CON; EV, EV-treated group; PAL; EP, EV+palmitate-treated group. RESULTS: The palmitate-treated osteoblasts showed increased expression of inositol-requiring protein 1α (IRE1α), decreased sarcoplasmic/endoplasmic reticulum Ca2+-ATPase (SERCA), reduced ER Ca2+, and mitochondrial membrane potential (MMP), indicating induced ER stress. Palmitate-induced ER stress was associated with elevated nuclear factor kappa B (NFκB) and receptor activator of NFκB (RANKL) levels and an increased RANKL/osteoprotegerin (OPG) ratio, causing bone loss. The treatment of osteoblasts with EVs from muscle contraction was found to improve ER stress, the levels of ER Ca2+, MMP, NFκB, and RANKL, and the RANKL/OPG ratio. CONCLUSIONS: Palmitate reduces ER Ca2+ and induces osteoblast ER stress while impairing mitochondrial function. Furthermore, through the RANKL/RANK/OPG pathway, a key mechanism related to bone loss is induced. However, substances induced by muscle contraction can act on osteoblasts through exosomes, potentially improving this process.
참고문헌 (27)
- Qiao D, "Association of obesity with bone mineral density and osteoporosis in adults : a systematic review and meta-analysis", Public Health, 180, pp.22~28 (2020)
- Kim S, "Risk factors of bone mass loss at the lumbar spine : a longitudinal study in healthy Korean pre-and perimenopausal women older than 40 years", PloS One, 10(8), pp.e0136283~ (2015)
- Xu S, "Palmitate induces ER calcium depletion and apoptosis in mouse podocytes subsequent to mitochondrial oxidative stress", Cell Death Dis, 6(11), pp.e1976~ (2015)
- Yuzefovych L, "Different effects of oleate vs. palmitate on mitochondrial function, apoptosis, and insulin signaling in L6 skeletal muscle cells: role of oxidative stress", Am J Physiol Endocrinol Metab, 299(6), pp.1096~1105 (2010)
- Li H, "Defective autophagy in osteoblasts induces endoplasmic reticulum stress and causes remarkable bone loss", Autophagy, 14(10), pp.1726~1741 (2018)
- Min SK, "Position statement : exercise guidelines to increase peak bone mass in adolescents", Exercise Science, 29(1), pp.10~23 (2020)
- Kang YS, "Effects of swimming exercise on serum irisin and bone FNDC5 in rat models of high-fat diet-induced osteoporosis", J Sports Sci Med, 18(4), pp.596~ (2019)
- Hart K, "Swim-trained rats have greater bone mass, density, strength, and dynamics", J Appl Physiol, 91(4), pp.1663~1668 (2001)
- Falcai MJ, "The osteogenic effects of swimming, jumping, and vibration on the protection of bone quality from disuse bone loss", Scand J Med Sci Sports, 25(3), pp.390~397 (2015)
- Kawanishi N, "Electrical pulse stimulation-induced muscle contraction alters the microRNA and mRNA profiles of circulating extracellular vesicles in mice", Am J Physiol Regul Integr Comp Physiol, 324(6), pp.761~771 (2023)
- Raposo G, "Extracellular vesicles : exosomes, microvesicles, and friends", J Cell Biol, 200(4), pp.373~383 (2013)
- Vechetti Jr IJ, "The role of extracellular vesicles in skeletal muscle and systematic adaptation to exercise", J Physiol, 599(3), pp.845~861 (2021)
- Tobeiha M, "RANKL", RANK, Biomed Res Int, pp.2020[DOI]
- Laybutt D, "Endoplasmic reticulum stress contributes to beta cell apoptosis in type 2 diabetes", Diabetologia, 50, pp.752~763 (2007)
- Lu J, "Palmitate causes endoplasmic reticulum stress and apoptosis in human mesenchymal stem cells : prevention by AMPK activator", Endocrinology, 153(11), pp.5275~5284 (2012)
- Berridge MJ, "The versatility and universality of calcium signalling", Nat Rev Mol Cell Biol, 1(1), pp.11~21 (2000)
- Luciani DS, "Roles of IP3R and RyR Ca2+ channels in endoplasmic reticulum stress and β-cell death", Diabetes, 58(2), pp.422~432 (2009)
- Periasamy M, "SERCA pump isoforms : their role in calcium transport and disease", Muscle Nerve, 35(4), pp.430~442 (2007)
- Panov AV, "Influence of calcium on NADH and succinate oxidation by rat heart submitochondrial particles", Arch Biochem Biophys, 316(2), pp.815~820 (1995)
- Gupta S, "HSP72 protects cells from ER stress-induced apoptosis via enhancement of IRE1α-XBP1signaling through a physical interaction", PLoS Biol, 8(7), pp.e1000410~ (2010)
- Gupta S, "The mitochondrial death pathway : a promising therapeutic target in diseases", J Cell Mol Med, 13(6), pp.1004~1033 (2009)
- Guo J, "PERK controls bone homeostasis through the regulation of osteoclast differentiation and function", Cell Death Dis, 11(10), pp.847~ (2020)
- Zhong X, "Bezafibrate prevents palmitate-induced apoptosis in osteoblastic MC3T3-E1 cells through the NF-κB signaling pathway", Int J Mol Med, 28(4), pp.535~542 (2011)
- Thyfault JP, "Exercise and metabolic health : beyond skeletal muscle", Diabetologia, 63, pp.1464~1474 (2020)
- Rome S, "Skeletal muscle-released extracellular vesicles : state of the art", Front Physiol, 10, pp.929~ (2019)
- Lawson C, "Microvesicles and exosomes : new players in metabolic and cardiovascular disease", J Endocrinol, 228(2), pp.57~71 (2016)
- Bakthisaran R, "Small heat shock proteins : role in cellular functions and pathology", Biochim Biophys Acta Proteins Proteom, 1854(4), pp.291~319 (2015)