Effects of bergapten on damages of osteocytes MLO-Y4 induced by TCP wear particles and its mechanism

doi:10.12047/j.cjap.5845.2019.124

Abstract

Abstract: Objective: To study the effects of bergapten (BP) on damages of osteocytes MLO-Y4 induced by tricalcium phosphate (TCP) wear particles and its mechanism. Methods: MLO-Y4 cells were treated with TCP wear particles for 48 h to establish the model of osteocytes injuries in vitro. The MLO-Y4 cells were divided into the following five groups: control group, TCP wear particles treated (0.1 mg/ml) group, bergapten (1, 5 and 20 μmol/L) treated groups. MTT assay and Calcein-AM staining were used to determine the viability of MLO-Y4 cells; Hoechst 33342 staining and the flow cytometry were applied to detect the apoptosis of MLO-Y4; real-time PCR was performed to examine the mRNA levels of dentin matrix protein1 (DMP-1), sclerostin (SOST) and fibroblast growth factor23 (FGF23); Western blot was performed to examine protein expressions of glucose-regulated protein 78 (GRP78), protein kinase R-like ER kinase (PERK) phospho-PERK (p-PERK), eukaryotic initiation factor 2α (eIF2α), phospho-eIF2α (p-eIF2α), activating transcription factor 4 (AFT4), C/EBP homologous protein (CHOP) and caspase-3 in MLO-Y4 cells. Results: Compared with control group, the MLO-Y4 viability and DMP-1 mRNA level in TCP group were decreased significantly (P＜0.05), while the percentage of apoptosis and mRNA levels of SOST and FGF23 were obviously increased (P＜0.05), and protein expressions of GRP78, AFT4, CHOP, p-PERK/PERK and p-eIF2α/eIF2α were up-regulated significantly in MLO-Y4 cells (P＜0.05). Compared with TCP group, the damages of MLO-Y4 and cell apoptosis in bergapten treated groups were decrease obviously (P＜0.05), the expressions of GRP78, AFT4, CHOP, p-PERK/PERK and p-eIF2α/eIF2α were down-regulated remarkably (P＜0.05). Conclusion: Bergapten can inhibit osteocytes damages induced by TCP wear particles, which may be related to reducing ER stress and PERK pathway activation.

Key words: bergapten, TCP wear particles, osteocytes, apoptosis, ER stress

CLC Number:

R363.3

YANG Zi-jian, HUANG Jun-yao, GAO Ye-fei, HUANG Wen-ji, XU Shi-lei, JIN Jing-jing, WAN Ye-dong, YAN Ming, MAO Hong-jiao, ZHANG Yun. Effects of bergapten on damages of osteocytes MLO-Y4 induced by TCP wear particles and its mechanism[J]. CJAP, 2019, 35(6): 563-568.

References

[1] Dumbleton JH, Manley MT, Edidin AA. A literature review of the association between wear rate and osteolysis in total hip arthroplasty [J]. J Arthroplasty, 2002, 17(5): 649-661.
[2] Stea S, Visentin M, Granchi D, et al. Cytokines and osteolysis around total hip prostheses [J]. Cytokine, 2000, 12(10): 1575-1579.
[3] Ormsby RT, Cantley M, Kogawa M, et al. Evidence that osteocyte perilacunar remodelling contributes to polyethylene wear particle induced osteolysis [J]. Acta Biomater, 2016, 33: 242-251.
[4] 严嘉琦, 张云, 刘方舒, 等. 骨细胞在TCP磨损颗粒诱导假体周围骨溶解中的作用[J]. 中国应用生理学杂志, 2018, 34(1): 80-85.
[5] Wang R, Wang Z, Ma Y, et al. Particle-induced osteolysis mediated by endoplasmic reticulum stress in prosthesis loosening [J]. Biomaterials, 2013, 34(11): 2611-2623.
[6] Follet H, Li J, Phipps RJ, et al. Risedronate and alendronate suppress osteocyte apoptosis following cyclic fatigue loading [J]. Bone, 2007, 40(4): 1172-1177.
[7] Ye T, Cao P, Qi J, et al. Protective effect of low-dose risedronate against osteocyte apoptosis and bone loss in ovariectomized rats [J]. PLoS One, 2017, 12(10): e0186012.
[8] Komm BS, Morgenstern D, Yamamoto LA, et al. The safety and tolerability profile of therapies for the prevention and treatment of osteoporosis in postmenopausal women [J]. Expert Rev Clin Pharmacol, 2015, 8(6): 769-784.
[9] Yang Y, Zheng K, Mei W, et al. Anti-inflammatory and proresolution activities of bergapten isolated from the roots of Ficus hirta in an in vivo zebrafish model [J]. Biochem Biophys Res Commun, 2018, 496(2): 763-769.
[10]Pattanayak SP, Bose P, Sunita P, et al. Bergapten inhibits liver carcinogenesis by modulating LXR/PI3K/Akt and IDOL/LDLR pathway [J]. Biomed Pharmacother, 2018, 108: 297-308.
[11]Xiao JJ, Zhao WJ, Zhang XT, et al. Bergapten promotes bone marrow stromal cell differentiation into osteoblasts in vitro and in vivo [J]. Mol Cell Biochem, 2015, 409(1-2): 113-122.
[12]Zheng M, Ge Y, Zhang Y, et al. Bergapten prevents lipopolysaccharide mediated osteoclast formation, bone resorption and osteoclast survival [J]. Int Orthop, 2014; 38(3): 627-634.
[13]Chen G, Xu Q, Dai M, et al. Bergapten suppresses RANKL-induced osteoclastogenesis and ovariectomy-induced osteoporosis via suppression of NF-κB and JNK signaling pathways [J]. Biochem Biophy Res Commun, 2019, 506(2): 329-334.
[14]Zawawi MS, Marino V, Perilli E, et al. Parthenolide reduces empty lacunae and osteoclastic bone surface resorption induced by polyethylene particles in a murine calvarial model of peri-implant osteolysis [J]. J Biomed Mater Res A, 2015, 103(11): 3572-3579.
[15]Yoshiko Y, Wang H, Minamizaki T, et al. Mineralized tissue cells are a principal source of FGF23 [J]. Bone, 2007, 40(6): 1565-1573.
[16]Zhang Y, Yan M, Yu A, et al. Inhibitory effects of β-tricalciumphosphate wear particles on osteocytes via apoptotic response and Akt inactivation [J]. Toxicology, 2012, 297(1-3): 57-67.
[17]Sato AY,Tu X, McAndrews KA, et al. Prevention of glucocorticoid induced-apoptosis of osteoblasts and osteocytes by protecting against endoplasmic reticulum (ER) stress in vitro and in vivo in female mice [J]. Bone, 2015, 73: 60-68.
[18]Yin J, Han L, Cong W.Alpinumisoflavone rescues glucocorticoid-induced apoptosis of osteocytes via suppressing Nox2-dependent ROS generation [J]. Pharmacol Rep, 2018, 70(2): 270-276.
[19]赵明, 刘晓翠, 崔晓雪, 等. miRNA378^*表达对柯萨奇B3病毒感染乳鼠心肌细胞凋亡、网腔钙结合蛋白、内质网应激伴侣蛋白表达的影响[J]. 中国应用生理学杂志, 2017, 33(4): 304-307.
[20]Yin H, Zhao L, Jiang X, et al. DEV induce autophagy via the endoplasmic reticulum stress related unfolded protein response [J]. PLoS One, 2017, 12(12): e0189704.
[21]张晶晶, CHEN Jun-hao, 王万铁, 等. 内质网应激在大鼠低氧高二氧化碳性肺动脉高压中的作用[J]. 中国应用生理学杂志, 2018, 34(4): 324-333.
[22]Bonnet-Magnaval F, Philippe C, Van Den Berghe L, et al. Hypoxia and ER stress promote Staufen1 expression through an alternative translation mechanism [J]. Biochem Biophys Res Commun, 2016, 479(2): 365-371.
[23]Kang X, Yang W, Wang R, et al. Sirtuin-1 (SIRT1) stimulates growth-plate chondrogenesis by attenuating the PERK-eIF-2α-CHOP pathway in the unfolded protein response [J]. J Biol Chem, 2018, 293(22): 8614-8625.