miR-214-3p在骨质疏松大鼠弓状核表达变化中的初步研究

Study on changes of miR-214-3p expression in the arcuate nucleus of osteoporotic rats

摘要:

目的 研究miR-214-3p在骨质疏松大鼠弓状核的表达，初步揭示其在骨质疏松发生发展中的作用。方法将4月龄雌性SD大鼠随机分为实验组和对照组，每组6只，实验组行切除卵巢手术，对照组行假手术，常规饲养8周。应用苏木精-伊红染色法和骨形态计量学检测大鼠胫骨干骺端骨密度；生物信息学分析弓状核miR-214-3p作用的靶基因；荧光定量聚合酶链反应法测定大鼠弓状核阿黑皮素原mRNA表达水平以及弓状核、血清miR-214-3p表达水平；免疫组织化学染色法测定大鼠下丘脑弓状核阿黑皮素原表达强度。结果 与对照组比较，实验组大鼠胫骨干骺端骨小梁数量、骨小梁厚度和骨小梁面积百分比均明显减小（均P<0.01），骨小梁分离度明显升高（P<0.01）；实验组弓状核miR-214-3p表达水平升高（P<0.05），同时血清miR-214-3p表达水平明显升高（P<0.01）；生物信息学分析显示miR-214-3p与阿黑皮素原具有良好的靶向关系；与对照组相比较，实验组弓状核阿黑皮素原m RNA的表达水平降低（P<0.05），弓状核阿黑皮素原蛋白表达强度明显降低（P<0.01）。结论 骨质疏松大鼠弓状核miR-214-3p表达水平升高，可通过其自身或调控阿黑皮素原影响骨质疏松的发生发展。

Abstract:

Objective To investigate the expression of miR-214-3p in the arcuate nucleus of osteoporotic rats its role in the development of osteoporosis. Methods 4-month-old female SD rats were randomly divided into experimental and control groups, 6 rats in each group. The rats in experimental group and the rats in control group were treated with ovariectomy and sham operation, respectively for indicated times. Hematoxylin-Eosin (HE) staining and bone morphometrics were used to detect the bone density of rat tibial metaphysis. Bioinformatics analysis was used to analyze the targeted gene of miR-214-3p in the arcuate nucleus. RT-qPCR was used to determine the expression level of proopiomelanocortin (POMC) mRNA in the arcuate nucleus, as well as the expression level of miR-214-3p in the arcuate nucleus and serum. Immunohistochemistry assay was used to determine the expression level of POMC in the arcuate nucleus of rats. Results The number thickness and the percentage of trabecular area in the tibial metaphysis of rats in the experimental group were all significantly reduced (all P<0.01), whereas the separation of trabeculae was significantly increased (P<0.01), compared with controls. The expression levels of miR-214-3p in the arcuate nucleus of the experimental group were increased (P<0.05), while the expression levels of miR-214-3p in the serum (P<0.01) were also significantly increased. The result of bioinformatic analysis showed that miR-214-3p had a good targeting relationship with POMC. Compared with the control group, the expression levels of POMC mRNA in the arcuate nucleus were reduced in the experimental group (P<0.05), and the intensity of POMC expression in the arcuate nucleus was significantly reduced (P<0.01). Conclusion The results showed that elevated expression levels of miR-214-3p in the arcuate nucleus of osteoporotic rats, which would affect the development of osteoporosis by itself or its regulation for POMC.