Effects of bisphosphonates on the expression of Bcl-2, Bax and Caspase-3 in condylar cartilage of osteoporosis rats

doi:10.3969/j.issn.2095-4344.2016.27.003

Abstract

Abstract:

BACKGROUND: Bisphosphonates are a kind of traditional antiresorptive drugs, which may influence the growth and development of cartilage tissue. In recent years, there are many data describing how bisphosphonates affect joint cartilage of long bone, but their effects on condylar cartilage remain unclear.
OBJECTIVE: To clarify the effects of the third-generation bisphosphonate risedonate on the expression of Bcl-2 (anti-apoptotic), Bax (pro-apoptotic) and caspase-3 in the condylar cartilage of osteoporosis rats.
METHODS: Thirty female Sprague-Dawley rats were randomly divided into three groups. In the sham operation group, ovary was exposed but not resected. The model group received a bilateral ovariectomy to establish the models of osteoporosis, and then received saline solution treatment (2.4 μg/kg) subcutaneously every 3 days, once a day, from 3 days pre-surgery. The treatment group received a bilateral ovariectomy to establish the models of osteoporosis, and then received risedronate treatment (2.4 μg/kg) subcutaneously every 3 days from 3 days pre-surgery. Three months after the operation, the condylar cartilages of all animals were harvested. Apoptosis, the expression of Bcl-2, BAX and Caspase3 were observed.
RESULTS AND CONCLUSION: (1) The number of apoptotic cells in rat condylar cartilage and subcondylar region: the sham operation group < the treatment group < the model group (all P < 0.05). (2) Expression of Bcl-2: The trend of the model group was lower than that in the sham operation group, although there was no statistically significant difference between the two groups; Bcl-2 expression in the treatment group was statistically higher compared to the model group (P < 0.05). (3) Expression of Bax and Caspase-3: The expression levels of Bax and Caspase-3 were higher in the model group than in the sham operation group (all P < 0.05), while Bax and Caspase-3 expression was lower in the treatment group than that in the model group (all P < 0.05). The results suggested that bisphophonates can regulate apoptosis in condylar cartilage from osteoporosis rats by changing the expression of Bcl-2, Bax and Caspase-3.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

Key words: Bisphosphonates, Osteoporosis, Animals, Laboratory

CLC Number:

R318

Zhou Qi, Wei Li, Jiang Li-ting, Li Ning, Gao Yi-ming. Effects of bisphosphonates on the expression of Bcl-2, Bax and Caspase-3 in condylar cartilage of osteoporosis rats[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(27): 3970-3976.

Figures/Tables 1

2.1 实验动物数量分析及造模成功的验证 30只大鼠均进入结果分析，无脱失值。与假手术组骨密度(406.89± 15.64) g/L相比，模型组骨密度(299.17±15.30) g/L明显降低(P < 0.05)，提示骨质疏松造模成功。 2.2 各组大鼠髁突软骨TUNEL染色结果 Ikeda等[1]将髁突软骨分成4层：纤维层、增殖细胞层、软骨细胞层、肥大软骨细胞层。TUNEL染色显示，假手术组髁突软骨中凋亡细胞多位于软骨细胞层及肥大层，软骨下骨组织中也可见凋亡的骨细胞散在分布；模型组凋亡细胞可见于增殖层、软骨细胞层及肥大软骨细胞层，在软骨下骨骨细胞中的分布也增多；治疗组凋亡细胞散在分布于软骨细胞层与肥大层；模型组髁突软骨及软骨下骨凋亡细胞较假手术组显著增多(P < 0.01)，治疗组髁突软骨与软骨下骨凋亡细胞数较模型组减少( P < 0.05)，见图1。 2.3 各组髁突软骨及软骨下骨Bcl-2、Bax、Caspase3分布及表达情况免疫组织化学染色结果显示，Bcl-2表达多位于各组髁突软骨增殖层、软骨细胞层中，肥大层亦可见其表达，免疫反应定位于细胞质中(图2)；Bax多位于各组髁突软骨细胞层、肥大层浅层及胞外基质中，棕黄色颗粒多定位于细胞质中(图2)；Caspase3表达可见于各组髁突软骨细胞层、肥大层浅层，多定位于细胞质中(图3)，胞核几乎不染色，在模型组增殖层亦可见Capase3表达。模型组Bax蛋白表达较假手术组显著升高(P < 0.05)，治疗组Bax蛋白表达较模型组下调(P < 0.05)；模型组Bcl-2蛋白较假手术组略有增多，但差异无显著性意义；治疗组Bcl-2蛋白表达较模型组显著上调(P < 0.05)，且Bcl-2/Bax相对表达量比值较模型组显著升高(P < 0.05)；模型组Caspase3蛋白表达最显著(P < 0.05)，在软骨细胞层及肥大层显著表达，而在假手术组和治疗组仅可见少量表达，见图4。"

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