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Fas- and Mitochondria-Mediated Signaling Pathway Involved in Osteoblast Apoptosis Induced by AlCl3

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Abstract

Aluminum (Al) is known to induce apoptosis of osteoblasts (OBs). However, the mechanism is not yet established. To investigate the apoptotic mechanism of OBs induced by aluminum trichloride (AlCl3), the primary OBs from the craniums of fetal Wistar rats were exposed to 0 mg/mL (control group, CG), 0.06 mg/mL (low-dose group, LG), 0.12 mg/mL (mid-dose group, MG), and 0.24 mg/mL (high-dose group, HG) AlCl3 for 24 h, respectively. We observed that AlCl3 induced OB apoptosis with the appearance of apoptotic morphology and increase of apoptosis rate. Additionally, AlCl3 treatment activated mitochondrial-mediated signaling pathway, accompanied by mitochondrial membrane potential (ΔΨm) depolarization, release of cytochrome c from the mitochondria to the cytoplasm, as well as survival signal-related factor caspase-9 and caspase-3 activation. AlCl3 exposure also activated Fas/Fas ligand signaling pathway, presented as Fas, Fas ligand, and Fas-associated death domain expression enhancement and caspase-8 activation, as well as the hydrolysis of Bid to truncated Bid, suggesting that the Fas-mediated signaling pathway might aggravate mitochondria-mediated OB apoptosis through hydrolyzing Bid. Furthermore, AlCl3 exposure inhibited Bcl-2 protein expression and increased the expressions of Bax, Bak, and Bim in varying degrees. These results indicated that AlCl3 exposure induced OB apoptosis through activating Fas- and mitochondria-mediated signaling pathway and disrupted B-cell lymphoma-2 family proteins.

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Abbreviations

Al:

Aluminum

AlCl3 :

Aluminum trichloride

ANOVA:

One-way analysis of variance

AO/EB:

Acridine orange/ethidium bromide

Bcl-2:

B-cell lymphoma-2

Cyt-C:

Cytochrome c

DMEM:

Dulbecco’s minimum essential medium

FADD:

Fas-associated death domain

FasL:

Fas ligand

FBS:

Fetal bovine serum

JC-1:

5,5′,6,6′-Tetrachloro-1.1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide

MPTP:

Mitochondrial permeability transition pore

MTP:

Mitochondrial transmembrane potential

OBs:

Osteoblasts

qRT-PCR:

Quantitative real-time reverse transcription-polymerase chain reaction

SDS-PAGE:

SDS-polyacrylamide gel electrophoresis

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Funding

The study was supported by grants from the Natural Science Foundation of Heilongjiang Province (C201425) and the National Science Foundation Project (31372496).

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Authors and Affiliations

  1. Key Laboratory of the Provincial Education Department of Heilongjiang for Common Animal Disease Prevention and Treatment, College of Veterinary Medicine, Northeast Agricultural University, No. 59 Mucai Street, Xiangfang District, Harbin, 150030, China

    Feibo Xu, Miao Song, Bing Shao, Yanfei Han, Zheng Cao & Yanfei Li

  2. Muyuan Foodstuff Co., Ltd, Nanyang, 473000, China

    Limin Ren

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  1. Feibo Xu
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  2. Limin Ren
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  3. Miao Song
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  7. Yanfei Li
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Correspondence to Yanfei Li.

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Xu, F., Ren, L., Song, M. et al. Fas- and Mitochondria-Mediated Signaling Pathway Involved in Osteoblast Apoptosis Induced by AlCl3 . Biol Trace Elem Res 184, 173–185 (2018). https://doi.org/10.1007/s12011-017-1176-y

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