过表达circRNA_079813牙髓干细胞对大鼠牙周骨缺损的影响  

Influence of overexpressed circRNA_079813 dental pulp stem cells on periodontal bone defect in rats

作　　者：周洋[1] 帕米拉·甫拉提 刘景[1] ZHOU Yang;Pamila FULATI;LIU Jing(Department of Stomatology,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region 83001l,China)

机构地区：[1]新疆医科大学第五附属医院口腔科,新疆维吾尔自治区乌鲁木齐830011

出　　处：《中华实用诊断与治疗杂志》2023年第10期1009-1014,共6页Journal of Chinese Practical Diagnosis and Therapy

基　　金：新疆维吾尔自治区自然科学基金(2021D01C431)。

摘　　要：目的探讨过表达circRNA_079813的牙髓干细胞(DPSCs)对牙周骨缺损的修复作用及可能机制。方法对数生长期大鼠DPSCs分为对照组(正常培养基培养)与成骨诱导组(成骨诱导培养基培养),过表达组(转染pcDNA3.1-circRNA_079813过表达质粒)与空载组(转染pcDNA3.1空载质粒)。50只雄性SD大鼠随机分为假手术组、模型组、DPSCs组、pcDNA-null组、pcDNA-circ组各10只。模型组、DPSCs组、pcDNA-null组、pcDNA-circ组采用低速球钻在牙槽骨骨面作小切口制作牙周骨缺损模型,分别在缺损区域注入α-MEM培养基、DPSCs、空载组DPSCs、过表达组DPSCs;假手术组仅暴露牙槽骨骨面,不作后续处理。采用碱性磷酸酶(ALP)活性检测试剂盒检测各组DPSCs和各组大鼠牙周缺损骨组织ALP活性,采用实时荧光定量PCR法检测各组DPSCs和各组大鼠牙周缺损骨组织ALP mRNA、circRNA_079813相对表达量,采用ELISA法检测各组大鼠牙周缺损骨组织白细胞介素(IL)-1β、IL-6、肿瘤坏死因子-α(TNF-α)水平,采用Western blot法检测各组大鼠牙周缺损骨组织骨形态发生蛋白2(BMP-2)、Runt相关转录因子2(Runx2)、骨钙蛋白(OCN)相对表达量。结果(1)成骨诱导组ALP活性(5.88±0.19)、ALP mRNA相对表达量(2.19±0.30)均高于对照组(1.00±0.06、1.00±0.09)(t=42.421,P<0.001;t=6.581,P=0.003),circRNA_079813相对表达量(0.22±0.04)低于对照组(1.00±0.10)(t=12.544,P<0.001)。(2)过表达组ALP活性及ALP mRNA、circRNA_079813相对表达量(3.25±0.04、9.45±0.52、12.08±0.93)均高于空载组(1.00±0.12、1.00±0.02、1.00±0.10)(P<0.05)。(3)pcDNA-circ组牙周缺损骨组织circRNA_079813相对表达量(7.06±0.92)均高于假手术组、模型组、DPSCs组、pcDNA-null组(1.00±0.07、1.06±0.25、1.01±0.14、1.02±0.08)(P<0.05),假手术组、模型组、DPSCs组、pcDNA-null组两两比较差异均无统计学意义(P>0.05)。(4)DPSCs组、pcDNA-null组、pcDNA-circ组牙周缺损骨组织ALP活性(1.25±0.18、1.24±0.42Objective To investigate the role of dental pulp stem cells(DPSCs)with overexpressed circRNA_079813 in repairing periodontal bone defect in rats and the potential mechanisms.Methods The rat DPSCs in logarithmic growth phase were divided into control group(cultured in normal medium),osteogenic induction group(cultured in osteogenic induction medium),overexpression group(transfected with pcDNA3.1-circRNA_079813 overexpression plasmid)and null-loading group(transfected with pcDNA3.1 null-loading plasmid).Fifty male SD rats were randomly divided into sham-operation group,model group,DPSCs group,pcDNA-null group and pcDNA-circ group,with 10 rats in each group.Small incisions were made in the bone surface of the alveolar bone using a low-speed ball drill to prepare periodontal bone defect models,and α-MEM medium,DPSCs,DPSCs in null-loading group,and DPSCs in overexpression group were injected into the defect areas of model group,DPSCs group,pcDNA-null group and pcDNA-circ group,respectively.In sham-operation group,the bone surface of alveolar bone was only exposed without any subsequent treatment.The alkaline phosphatase(ALP)activity test kit was used to measure the ALP activity of DPSCs and periodontal defect bone tissues.Real-time fluorescence quantitative PCR was used to detect the relative expressions of ALP mRNA and circRNA_079813 in DPSCs and periodontal defect bone tissues.ELISA was used to detect the levels of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in periodontal defect bone tissues.Western blot was used to detect the relative expressions of bone morphogenetic protein(BMP)-2,Runt-related transcription factor2(Runx2),and osteocalcin(OCN)in periodontal defect bone tissues.Results(1)The ALP activity and relative expression of ALP mRNA were higher in osteogenic induction group(5.88±0.19,2.19±0.30)than those in control group(1.00±0.06,1.00±0.09)(t=42.421,P<0.001;t=6,581,P=0.003),while the relative expression of circRNA_079813 was lower in steogenic induction group(0.22±0.04)than that in c

关 键 词：circRNA_079813 牙髓干细胞 牙周骨缺损 成骨分化 大鼠 

分 类 号：R781[医药卫生—口腔医学]