Department of Orthopedics, The Second Xiangya Hospital of Central South University, Changsha, China.
Eur Rev Med Pharmacol Sci. 2020 Feb;24(3):1023-1029. doi: 10.26355/eurrev_202002_20152.
To study the influence of micro ribonucleic acid (miR)-137 on osteoporosis rats by regulating runt-related transcription factor 2 (RUNX2).
A total of 36 Sprague-Dawley rats were randomly assigned to the normal group (n=12), model group (n=12), and inhibitor group (n=12). No treatment was performed in the normal group. The osteoporosis model in rats was prepared in the model group, and miR-137 inhibitor was administered in osteoporosis rats of inhibitor group. Following 12 weeks of intervention, sampling was conducted. The expression of RUNX2 was detected via immunohistochemistry, and its protein expression level was determined via Western blotting. Quantitative Polymerase Chain Reaction (qPCR) was carried out to detect the mRNA level of miR-137. The contents of serum bone Gla protein (BGP) and total alkaline phosphatase (TALP) were measured using enzyme-linked immunosorbent assay (ELISA). Finally, bone mineral density was determined with a dual-energy X-ray absorptiometry instrument.
According to the immunohistochemistry detection, the rats in model group and inhibitor group had a notably lower positive expression level of RUNX2 than normal group (p<0.05), and its expression level in the inhibitor group was substantially higher than that in the model group (p<0.05). Western blotting results showed that compared with that in the normal group, the protein expression level of RUNX2 was notably lowered in the model and inhibitor group (p<0.05), which was markedly higher in the inhibitor group than that in the model group (p<0.05). It was found through the qPCR that the expression level of miR-137 was remarkably raised in both model group and inhibitor group compared with that in the normal group, showing statistically significant differences (p<0.05). The rats in the inhibitor group had a remarkably lower expression level of miR-137 than the model group (p<0.05). ELISA results revealed that the model group and inhibitor group had substantially lower contents of serum BGP and TALP than the normal group (p<0.05), and that their contents rose dramatically in the inhibitor group compared with that in the model group (p<0.05). Additionally, based on the measurement of bone mineral density, compared with that in the normal group, bone mineral density declined considerably in the model group and inhibitor group (p<0.05). It was markedly elevated in inhibitor group in comparison with that in the model group (p<0.05).
MiR-137 regulates RUNX2 to affect the bone mineral density of osteoporosis model rats.
通过调控 runt 相关转录因子 2(RUNX2)研究微小核糖核酸(miR)-137 对骨质疏松大鼠的影响。
36 只 Sprague-Dawley 大鼠随机分为正常组(n=12)、模型组(n=12)和抑制剂组(n=12)。正常组不做处理,模型组建立骨质疏松大鼠模型,抑制剂组给予骨质疏松大鼠 miR-137 抑制剂。干预 12 周后取材,采用免疫组织化学法检测 RUNX2 的表达,采用 Western blot 法检测其蛋白表达水平,采用实时荧光定量聚合酶链反应(qPCR)法检测 miR-137 的 mRNA 水平,采用酶联免疫吸附试验(ELISA)法检测血清骨钙素(BGP)和总碱性磷酸酶(TALP)含量,最后采用双能 X 线吸收仪测定骨密度。
免疫组织化学检测结果显示,模型组和抑制剂组大鼠 RUNX2 阳性表达水平明显低于正常组(p<0.05),抑制剂组明显高于模型组(p<0.05)。Western blot 结果显示,与正常组比较,模型组和抑制剂组 RUNX2 蛋白表达水平明显降低(p<0.05),抑制剂组明显高于模型组(p<0.05)。qPCR 结果显示,模型组和抑制剂组 miR-137 表达水平明显高于正常组,差异有统计学意义(p<0.05),抑制剂组明显低于模型组(p<0.05)。ELISA 结果显示,模型组和抑制剂组血清 BGP 和 TALP 含量明显低于正常组(p<0.05),抑制剂组明显高于模型组(p<0.05)。双能 X 线吸收仪检测结果显示,与正常组比较,模型组和抑制剂组骨密度明显降低(p<0.05),抑制剂组明显高于模型组(p<0.05)。
miR-137 通过调控 RUNX2 影响骨质疏松模型大鼠的骨密度。