lncRNA SNHG8抑制颌骨骨髓间充质干细胞成骨分化功能  

LncRNA SNHG8 inhibits osteogenic differentiation of jaw bone marrow mesenchymal stem cells

作　　者：刁展秋 刘华 杨昊清 刘惠娜[1] 范志朋[1] DIAO Zhanqiu;LIU Hua;YANG Haoqing;LIU Huina;FAN Zhipeng(Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京口腔医院,北京100050

出　　处：《口腔生物医学》2024年第4期185-192,共8页Oral Biomedicine

基　　金：国家自然科学基金(82100970)。

摘　　要：目的:研究lncRNA SNHG8对颌骨骨髓间充质干细胞(JBMMSCs)成骨分化的影响。方法:通过慢病毒转染敲除或过表达JBMMSCs中lncRNA SNHG8,实时荧光定量PCR检测基因表达,碱性磷酸酶(ALP)活性测定、茜素红染色、钙离子定量以及Western blot检测JBMMSCs的体外成骨分化能力,活性氧(ROS)染色检测ROS含量;lncRNA SNHG8敲低后的JBMMSCs与HA/TCP材料混合并植入裸鼠皮下,苏木素-伊红(HE)染色、Masson染色、免疫荧光染色检测JBMMSCs的体内成骨分化能力。结果:体外JBMMSCs中lncRNA SNHG8敲低后的ALP活性及体外矿化能力增强(P<0.01),骨涎蛋白(BSP)蛋白表达条带增强,lncRNA SNHG8过表达后ALP活性及体外矿化能力减弱(P<0.01),BSP蛋白表达条带减弱;敲除lncRNA SNHG8可上调MAPK通路中磷酸化c-Jun氨基末端激酶(p-JNK),过表达SNHG8可抑制p-JNK表达;敲除lncRNA SNHG8可增强胞内ROS染色,过表达lncRNA SNHG8可抑制胞内ROS染色。lncRNA SNHG8敲除后的体内新骨形成增加(P<0.01),BSP、骨钙素(OCN)蛋白表达条带增强。结论:lncRNA SNHG8可通过JNK信号通路抑制JBMMSCs体内外成骨分化功能。Objective:To investigate the effect of lncRNA small nucleolar RNA host genes(SNHG)8 on osteogenic differentiation of jaw bone marrow mesenchymal stem cells(JBMMSCs).Methods:LncRNA SNHG8 was eliminated or overexpressed in JBMMSCs by lentivirus transfection;gene expression was detected by qRT-PCR;alkaline phosphatase(ALP)activity detection,alizarinred staining,calcium ion quantification and Western blot were used to determine the osteogenic differentiation ability of JBMMSCs in vitro and the osteogenic differentiation ability of JBMMSCs in vivo was detected by hematoxylin-eosin(HE)staining.JBMMSCs with eliminated lncRNA SNHG8 were mixed with HA/TCP materials and subcutaneous transplanted in nude mice,Masson staining and immunofluorescence staining;reactive oxygen species(ROS)staining was used to detect ROS levels;and the protein expression of the mitogen-activated protein kinase(MAPK)signaling pathway was detected by Western blot.Results:After lncRNA SNHG8 was knocked down,ALP activity and in vitro mineralization ability of JBMMSCs were increased(P<0.01),and bone sialprotein(BSP)protein expression bands were increased.After lncRNA SNHG8 was overexpressed,ALP activity and in vitro mineralization ability were decreased(P<0.01),and BSP protein expression bands were decreased.The expression of phosphorylated c-Jun N-terminal kinase(p-JNK)in MAPK pathway was up-regulated by knocking down of lncRNA SNHG8,and overexpression of lncRNA SNHG8 inhibited p-JNK expression.LncRNA SNHG8 deletion can enhance intracellular ROS staining,and overexpression of lncRNA SNHG8 can inhibit intracellular ROS staining.After lncRNA SNHG8 knockout,new bone formation increased in vivo(P<0.01),and the expression bands of BSP and osteocalcin(OCN)were enhanced.Conclusions:LncRNA SNHG8 may inhibit the osteogenic differentiation function of JBMMSCs in vitro and in vivo via JNK signaling pathway.

关 键 词：颌骨骨髓间充质干细胞 lncRNA SNHG8 成骨分化 活性氧 C-JUN氨基末端激酶 

分 类 号：R780.2[医药卫生—口腔医学]