长链非编码RNA-TCONS00041960对大鼠激素性股骨头坏死模型中成脂和成骨基因表达的影响  

Effect of long non-coding RNA-TCONS_00041960 on adipogenic and osteogenic gene expression in rat model of steroid-induced osteonecrosis of the femoral head

作　　者：周杨 刘华[2] 徐双燕 贾祯 孙肖雅 米洋 梅英武 王秀利[3] 刘柯希 王义生[3] 李月白 Zhou Yang;Liu Hua;Xu Shuangyan;Jia Zhen;Sun Xiaoya;Mi Yang;Mei Yingwu;Wang Xiuli;Liu Kexi;Wang Yisheng;Li Yuebai(Department of Biochemistry and Molecular Biology,Basic Medical College,Zhengzhou University,Zhengzhou 450001,China;Morphological Center,Basic Medical College,Zhengzhou University,Zhengzhou 450001,China;Department of Orthopaedics,First Affiliated Hospital of Zhengzhou University,Open Laboratory of Unode Science of Clinical Medicine of Henan Province,Zhengzhou 450052,China)

机构地区：[1]郑州大学基础医学院生物化学与分子生物学系,450001 [2]郑州大学基础医学院形态学中心,450001 [3]郑州大学第一附属医院骨科,河南省高等学校临床医学重点学科开放实验室,450052

出　　处：《中华实验外科杂志》2021年第9期1748-1751,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81772427)。

摘　　要：目的观察长链非编码RNA(lncRNA)-TCONS00041960在大鼠激素性股骨头坏死模型中对过氧化物酶体增殖物激活受体γ(PPARγ)和成骨基因runt相关转录因子2(Runx2)表达的调控作用。方法健康SD大鼠64只,随机分为4组:(1)正常对照组(Blank组)。每次臀肌注射2 ml生理盐水,连续3 d。(2)激素组(Dex组)。每次臀肌注射地塞米松20 mg/kg,连续3 d,制作大鼠激素性股骨头坏死模型。(3)无关序列组(Ex-NC+Dex组)。同法给予地塞米松,一侧股骨头内注入转染无关序列慢病毒载体的骨髓间充质干细胞(BMSCs)。(4)激素+重组慢病毒组(Ex-Lnc+Dex组)。同法给予地塞米松,一侧股骨头内注入转染重组lncRNA TCONS00041960 lncRNA载体的BMSCS。于实验第4、8周提取各组大鼠股骨头组织中总RNA及总蛋白,应用TaqMan实时定量聚合酶链反应(Real-time PCR)与蛋白质印迹法(Western blot)检测成脂基因PPARγ、成骨特异性转录因子Runx2 mRNA及其蛋白表达。两样本两组间比较采用t检验;数值变量资料统计采用方差分析,组间比较采用最小显著性差异法(LSD)。结果实验第4周,Ex-Lnc+Dex组股骨头细胞内PPARγ mRNA与蛋白呈低表达(1.129±0.024与0.366±0.007),与Blank组(1.001±0.055与0.352±0.003)接近,差异均无统计学意义(t4w-mRNA=3.052,P>0.05;t4w-蛋白=3.227,P>0.05);明显低于Dex组(2.351±0.086与0.836±0.032)、Ex-NC+Dex组(2.226±0.484与0.888±0.067),差异均有统计学意义(F4w-mRNA=296.841,P<0.01;F4w-蛋白=196.684,P<0.01)。Ex-Lnc+Dex组Runx2 mRNA与蛋白呈高表达(1.176±0.036与0.963±0.014),与Blank组(1.001±0.077与0.934±0.019)接近,差异均无统计学意义(t4w-mRNA=2.893,P>0.05;t4w-蛋白=2.154,P>0.05);明显高于Dex组(0.522±0.007与0.501±0.005)、Ex-NC+Dex组(0.614±0.003与0.521±0.008),差异均有统计学意义(F4w-mRNA=376.700,P<0.01;F4w-蛋白=2 239.834,P<0.01)。实验第8周各组表达量与第4周一致。结论重组lncRNA-TCONS00041960载体能够有效下调大鼠激素性股骨头�Objective To explore the effect of long non-coding RNA(lncRNA)-TCONS_00041960 on adipogenic and osteogenic genes expression in rat model of steroid-induced osteonecrosis of the femoral head.Methods Totally,64 healthy SD rats were randomly divided into 4 groups:(1)normal control group(blank group)(2 ml physiological saline was injected into gluteal muscle for 3 days).(2)hormone group(Dex group)[Dexamethasone(20 mg/kg)was injected into gluteal muscle for 3 days to establish steroid-induced femoral head necrosis model].(3)unrelated sequence group(Ex-NC+Dex group)[Dexamethasone was given in the same way,and bone marrow mesenchymal stem cells(BMSCs)transfected with lentivirus of unrelated sequence were injected into one side of femoral head].(4)Dexamethasone+recombinant lentivirus group(Ex-Lnc+Dex group)[Dexamethasone was given in the same way,and BMSCs transfected with recombinant lentivirus vector lncRNA-TCONS_00041960 were injected into one side of femoral head].At 4th and 8th week,total RNA and total protein were extracted from the femoral head tissue of each group.The expression of the peroxisome proliferator-activated receptor-γ(PPARγ),runt-related transcription factor 2(Runx2)mRNA and protein was detected by TaqMan real-time quantitative polymerase chain reaction(Real-time PCR)and Western blotting at 4th week and 8th week of the experiment.The comparison between the two groups was performed by t test;the numerical variable data statistics were performed by analysis of variance,and the comparison between groups was performed by the least significant difference method(LSD).Results At 4 weeks,the expression levels of PPARγmRNA and protein in the femoral head cells in the Ex-Lnc+Dex group(1.129±0.024 and 0.366±0.007)were not significantly different from those in the blank group(1.001±0.055 and 0.352±0.003,t4w-mRNA=3.052,P>0.05;t4w-protein=3.227,P>0.05),but significantly lower than those in the Dex group(2.351±0.086 and 0.836±0.032)and Ex-NC+Dex group(2.226±0.484 and 0.888±0.067,F4w-mRNA=296.841,P<0.01;F4

关 键 词：长链非编码RNA 激素性股骨头坏死 干细胞 基因表达 

分 类 号：R681.8[医药卫生—骨科学] R-332[医药卫生—外科学]