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

刁展秋 ¹刘华 ¹杨昊清 ¹刘惠娜 ¹范志朋¹

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作者信息

1. 首都医科大学附属北京口腔医院,北京 100050
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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.

Abstract

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 stai-ning,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 immunofluores-cence staining;reactive oxygen species(ROS)staining was used to detect ROS levels;and the protein expression of the mitogen-acti-vated 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 expres-sion.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

Key words

jaw bone marrow mesenchymal stem cells/lncRNA small nucleolar RNA host genes 8/osteogenic differentiation/re-active oxygen species/c-Jun N-terminal kinase

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基金项目

国家自然科学基金(82100970)

出版年

2024

口腔生物医学

南京医科大学

口腔生物医学

CSTPCD

影响因子：0.423

ISSN：1674-8603

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