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CJAP ›› 2019, Vol. 35 ›› Issue (6): 563-568.doi: 10.12047/j.cjap.5845.2019.124

• ORIGINAL ARTICLES • Previous Articles     Next Articles

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

YANG Zi-jian1, HUANG Jun-yao1, GAO Ye-fei1, HUANG Wen-ji1, XU Shi-lei1, JIN Jing-jing1, WAN Ye-dong1, YAN Ming2, MAO Hong-jiao1, ZHANG Yun1△   

  1. 1. College of Medicine, Shaoxing University, Shaoxing 312000;
    2. School of Automation, Biomedical Engineering Department, Hangzhou Dianzi University, Hangzhou 310018, China
  • Received:2019-03-14 Online:2019-11-28 Published:2020-04-02

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

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