Inhibitory effects of vitamin E on osteocyte apoptosis and DNA oxidative damage in bone marrow hemopoietic cells at early stage of steroid-induced femoral head necrosis

Jia,Yan‑Bo; Jiang,Dian‑Ming; Ren,Yi‑Zhong; Liang,Zi‑Hong; Zhao,Zhen‑Qun; Wang,Yu‑Xin

doi:10.3892/mmr.2017.6160

Inhibitory effects of vitamin E on osteocyte apoptosis and DNA oxidative damage in bone marrow hemopoietic cells at early stage of steroid-induced femoral head necrosis

Authors:
- Yan‑Bo Jia
- Dian‑Ming Jiang
- Yi‑Zhong Ren
- Zi‑Hong Liang
- Zhen‑Qun Zhao
- Yu‑Xin Wang
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Affiliations: Department of Orthopaedics, The First Affiliated Hospital of Chongqing Medical University, Yuanjia Gang, Chongqing 400016, P.R. China, Department of Orthopaedics, The Second Affiliated Hospital of Inner Mongolia Medical University, Huhhot, Inner Mongolia 010030, P.R. China, Department of Neurology, Inner Mongolia Autonomous Region People's Hospital, Huhhot, Inner Mongolia 010030, P.R. China
Published online on: February 2, 2017 https://doi.org/10.3892/mmr.2017.6160
Pages: 1585-1592
Copyright: © Jia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Apoptosis and DNA oxidative damage serve significant roles in the pathogenesis of steroid‑induced femoral head necrosis. Vitamin E demonstrates anti‑apoptotic and anti‑oxidant properties. Therefore, the present study investigated the effects of vitamin E on osteocyte apoptosis and DNA oxidative damage in bone marrow hemopoietic cells at an early stage of steroid‑induced femoral head osteonecrosis. Japanese white rabbits were randomly divided into three groups (steroid, vitamin E‑treated, and control groups), each comprising 12 rabbits. Those in the steroid group (group S) were initially injected twice with an intravenous dose of 100 µg/kg Escherichia coli endotoxin, with a 24 h interval between the two injections, and then with an intramuscular dose of 20 mg/kg methylprednisolone, three times at intervals of 24 h in order to establish a rabbit model of osteonecrosis. The vitamin E treated group (group E) received the same treatment as group S, and were administered 0.6 g/kg/d vitamin E daily from the beginning of modeling. The control group (group C) was injected with normal saline at the equivalent dosage and times as the aforementioned two groups. Two time points, weeks 4 and 6 following the completion of modeling, were selected. Osteonecrosis was verified by histopathology with hematoxylin-eosin staining. The apoptosis rate of osteonecrosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling assay. The apoptosis expression levels of caspase‑3 and B‑cell lymphoma 2 (Bcl‑2), and DNA oxidative damage of bone marrow hematopoietic cells were analyzed by immunohistochemistry. At weeks 4 and 6 following the completion of modeling, the vacant bone lacunae rates of group E were 15.87±1.97 and 25.09±2.67%, respectively, lower than the results of 20.02±2.21 and 27.79±1.39% for group S; and the osteocyte apoptosis indexes of group E were 20.99±2.95 and 33.93±1.62%, respectively, lower than the results of 26.46±3.37 and 39.90±3.74% from group S. In addition, the Bcl-2 expression at week 4 in the femoral head tissues of group E was higher compared with group S; and the proportion of Bcl‑2‑positive cells of group E was 9.81±1.01%, higher compared with group S at 8.26±1.13%. The caspase‑3 staining data at week 4 in femoral head tissues demonstrated that in the 12 femoral heads of group S, four were negative (32%) and eight were positive (68%); in group E, five were negative (45%) and seven were positive (55%); and in group C, 11 were negative (95%) and one was positive (5%). In addition, the DNA oxidative damage rate at week 4 in the bone marrow hemopoietic cells of group E was (7.24±1.44%), lower compared with group S (11.80±1.26%), and higher compared with group C (5.75±1.47%). Vitamin E is effective in intervening in apoptosis through decreasing caspase‑3 expression and upregulating Bcl‑2 expression, and by alleviating DNA oxidative damage in bone marrow hemopoietic cells at the early stage of steroid‑induced femoral head necrosis in rabbit models.

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