摘要
目的 探讨微小RNA (miR)-27a-5p对抑郁症大鼠桡骨骨折愈合的影响。方法 将SD大鼠随机分为骨折对照组、骨折+抑郁组、miR-27a-5p agomir(激活剂)阴性对照组、miR-27a-5p agomir组、miR-27a-5p antagomir(抑制剂)阴性对照组、miR-27a-5p antagomir组,除骨折对照组外,其余各组大鼠均通过慢性温和不可预知应激刺激建立抑郁模型。所有大鼠均折断前右肢桡骨中段制备骨折模型,并以miR-27a-5p激活剂及抑制剂进行干预分组后,以强迫游泳实验与蔗糖水消耗实验检测各组大鼠抑郁症状。检测各组大鼠骨折段骨痂量和骨微结构;检测各组大鼠血清炎性因子白细胞介素(IL)-2、IL-6及核因子κB受体活化因子配体(RANKL)、骨保护素(OPG)水平;采用实时荧光定量聚合酶链反应(qRT-PCR)实验检测各组大鼠骨痂组织miR-27a-5p、IL-2 mRNA表达。体外培养大鼠骨髓间充质干细胞(BMSC),随机分为对照组、miR-27a-5p mimics(模拟物)组、miR-27a-5p mimics阴性对照组、miR-27a-5p inhibitor(抑制剂)组、miR-27a-5p inhibitor阴性对照组。诱导各组细胞成骨分化并以miR-27a-5p模拟物及抑制剂分组干预后,以碱性磷酸酶(ALP)染色检测各组细胞成骨分化;以qRT-PCR实验检测各组细胞miR-27a-5p、IL-2 mRNA表达。以双荧光素酶报告基因实验检测大鼠BMSC中miR-27a-5p对IL-2的靶向调节作用。结果 与骨折对照组相比,骨折+抑郁组大鼠强迫游泳不动时间延长,血清IL-2、IL-6及RANKL水平、骨痂组织IL-2 mRNA表达升高,蔗糖水消耗量、骨痂量、骨体积分数、骨小梁厚度、骨小梁数量减少,血清OPG水平、骨痂组织miR-27a-5p表达降低,差异有统计学意义(P<0.05)。与骨折+抑郁组相比,miR-27a-5p agomir组大鼠强迫游泳不动时间缩短,血清IL-2、IL-6及RANKL水平、骨痂组织IL-2 mRNA表达降低,蔗糖水消耗量、骨痂量、骨体积分数、骨小梁厚度、骨小梁数量增加,血清OPG水平、骨痂组织miR-27a-5p表达升�
Objective To explore the influence of microRNA(miR)-27a-5p on healing of radial fractures in rats with depression. Methods The SD rats were randomly separated into fracture control group, fracture + depression group, miR-27a-5p agomir(activator) negative control group, miR-27a-5p agomir group, miR-27a-5p antagomir negative(inhibitor) control group, and miR-27a-5p antagomir group, except for the fracture control group, the rats in the other groups were all established depression models through chronic mild and unpredictable stress stimulation. All the rats were fractured the middle part of the radius of the front right limb to prepare the fracture model, and after intervention with miR-27a-5p activator and inhibitor, the depressive symptoms of rats in each group were measured by forced swimming test and sucrose water consumption test;the amount of callus and bone microstructureof the fractured segment of the rats in each group were measured. The levels of serum inflammatoryfactors interleukin(IL)-2, IL-6, receptor activator of nuclear factor kappa B ligand(RANKL) andosteoprotegerin(OPG) were measured in each group;the expressions of miR-27a-5p andIL-2mRNAin callus tissue of rats in each group were measured by real-time fluorescence quantitative( qRT-PCR) experiment. The rat bone marrow mesenchymal stem cell(BMSC) were culturedin vitroandrandomly divided into control group, miR-27a-5p mimics group, miR-27a-5p mimics negative con-trol group, and miR-27a-5p inhibitor group and miR-27a-5p inhibitor negative control group. Afterinduced osteogenic differentiation of cells in each group and intervened in groups with miR-27a-5pmimics and inhibitors, the osteogenic differentiation of cells in each group was measured by alkalinephosphatase(ALP) staining;the expression of miR-27a-5p andIL-2mRNA of cells in each groupwas measured by qRT-PCR experiment. The targeting regulation effect of miR-27a-5p on IL-2 in ratBMSC was measured by dual-luciferase reporter gene assay.Results Compared with the fracturecontrol group, the forced
作者
王振龙
刘勇
祝茂
叶章勇
WANG Zhenlong;LIU Yong;ZHU Mao;YE Zhangyong(Department of Orthopedics and Traumatology,Affiliated Hospital of Traditional Chinese Medicine of Southwest Medical University,Luzhou,Sichuan,646000;Department of Orthopedics and Traumatology,Xuyong County Hospital of Traditional Chinese Medicine of Luzhou City in Sichuan Province,Luzhou,Sichuan,646400)
出处
《实用临床医药杂志》
CAS
2023年第1期84-91,共8页
Journal of Clinical Medicine in Practice
基金
四川省泸州市科技计划项目[2017-S-41(1/7)]。
