LncRNA HAGLR激活RUNX2并抑制NLRP3炎症小体对胫骨骨折愈合的影响  

LncRNA HAGLR activates RUNX2 and inhibits NLRP3 inflammasome to promote tibial fracture healing

作　　者：王文[1] 陈新宇 黄兹艺 邓杨柳 崔红旺 Wang Wen;Chen Xinyu;Huang Ziyi;Deng Yangliu;Cui Hongwang(Emergency and Trauma Surgery,The First Affiliated Hospital of Hainan Medical College,Haikou 570100)

机构地区：[1]海南医学院第一附属医院急诊和创伤外科,海口570100

出　　处：《安徽医科大学学报》2023年第5期830-837,共8页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81760260)。

摘　　要：目的研究长链非编码RNA(LncRNA)HAGLR对胫骨骨折(TF)小鼠的NOD样受体蛋白3(NLRP3)炎症小体表达和骨折愈合的影响并探讨机制。方法体外对成骨细胞MC3T3-E1沉默HAGLR,CCK-8法检测细胞活力,TUNEL法检测细胞凋亡,qPCR法检测骨碱性磷酸酶(BALP)和骨钙素的表达。Western blot法检测RUNX2、磷酸化RUNX2(p-RUNX2)以及NLRP3、半胱天冬酶1(Caspase1)、凋亡相关斑点样蛋白(ASC)、白细胞介素-1β(IL-1β)的表达。通过小鼠TF手术建立TF小鼠模型,在模型小鼠体内过表达HAGLR,并在过表达HAGLR的基础上沉默RUNX2或加入炎症小体抑制剂MCC950。qPCR法检测HAGLR和RUNX2的表达,Western blot法检测胰岛素样生长因子-1(IGF-1)的表达。microCT测量小鼠骨痂体积(MBV),称量小鼠的全长胫骨湿重。结果沉默HAGLR导致MC3T3-E1细胞活力降低且凋亡率增加(P<0.05),且RUNX2、p-RUNX2、BALP和骨钙素表达量均降低(P<0.05),NLRP3、Caspase1、ASC、IL-1β的表达量都增加(P<0.05)。与健康组织比较,TF小鼠体内HAGLR和RUNX2表达量降低(P<0.05)。过表达HAGLR促进TF小鼠体内的HAGLR和RUNX2表达,并增加MBV和全长胫骨湿重以及IGF-1的表达量(P<0.05)。在过表达HAGLR的基础上沉默RUNX2导致TF小鼠的MBV、全长胫骨湿重和IGF-1表达量都降低(P<0.05)。而在过表达HAGLR的基础上加入NLRP3炎症小体抑制剂MCC950导致MBV、全长胫骨湿重和IGF-1表达又增加(P<0.05)。结论LncRNA HAGLR通过激活RUNX2并抑制NLRP3炎症小体促进TF的愈合。Objective To study the effect of long non-coding RNA(LncRNA)HAGLR on the expression of NOD-like receptor pyrin domain-associated protein 3(NLRP3)inflammasome and fracture healing in tibial fracture(TF)mice and to explore the mechanism.Methods First,HAGLR in osteoblast MC3T3-E1 was silenced by in vitro.Cell viability was detected by CCK-8 assay,cell apoptosis was detected by TUNEL assay,and the expressions of bone alkaline phosphatase(BALP)and osteocalcin were detected by qPCR.Western blot assay was used to detect the expressions of RUNX2,phosphorylated RUNX2(p-RUNX2),NLRP3,cysteine aspartic protease 1(Caspase1),apoptosis-associated spot-like protein(ASC)and interleukin-1β.TF mouse models were established by tibial fracture operation in mice.HAGLR was overexpressed in the model mice,and RUNX2 was silenced or an inflammatory body inhibitor MCC950 was added on the basis of overexpression of HAGLR.The expressions of HAGLR and RUNX2 were detected by qPCR,and the expressions of insulin-like growth factor(IGF-1)were detected by Western blot.microCT was used to measure the volume of mouse callus(MBV)and the total tibial wet weight.Results The apoptosis rate of MC3T3-E1 cells increased and the expression levels of RUNX2,p-RUNX2,BALP and osteocalcin decreased(P<0.05).The expressions of NLRP3,Caspase1,ASC and IL-1βincreased(P<0.05).Compared with healthy tissue,the expressions of HAGLR and RUNX2 in TF mice decreased.Overexpression of HAGLR promoted the expressions of HAGLR and RUNX2 in TF mice,and increased the expression of MBV and tibia wet weight and IGF-1(P<0.05).Silencing RUNX2 on the basis of overexpression of HAGLR resulted in decreased expression of MBV,tibial wet weight and IGF-1 in TF mice(P<0.05).However,the addition of NLRP3 inflammasome inhibitor MCC950 on top of the overexpression of HAGLR resulted in increased expressions of MBV,full-length tibia wet weight and IGF-1(P<0.05).Conclusion LncRNA HAGLR promotes the healing of tibial fractures by activating RUNX2 and inhibiting NLRP3 inflammasome.

关 键 词：胫骨骨折 同源框D基因簇反义生长相关的长非编码RNA Runt相关转录因子2 NOD样受体热蛋白结构域相关蛋白3 炎症小体 

分 类 号：R683[医药卫生—骨科学]