摘要
目的探讨神经调节蛋白4(Nrg4)及其受体ErbB4在牙周炎患者龈沟液(GCF)中的表达情况及对牙周炎大鼠牙周组织炎症和牙槽骨丢失的影响。方法招募健康志愿者及牙周炎患者80名,分为健康组(H组)、牙周炎组(G组)、Ⅱ期B级牙周炎组(P1组)和Ⅲ期B级牙周炎组(P2组),每组20例。采集GCF,采用ELISA法检测ErbB4和Nrg4的表达量。在体外使用IFNγ+LPS刺激原代骨髓来源巨噬细胞(BMDMs)和树突状细胞(BMDCs),并采用qPCR法测定胞内表皮生长因子受体(EGFR)、ErbB2、ErbB3、ErbB4 mRNA的表达情况。使用Nrg4重组蛋白处理骨髓来源M1巨噬细胞并采用CCK-8法测定其存活能力,采用Annexin V-PI双标记流式细胞术测定其凋亡情况。在体动物实验构建实验性牙周炎动物模型。15只大鼠随机分为假手术组(S组)、模型组(M组)和Nrg4重组蛋白治疗组(T组),每组5只。对3组大鼠上颌骨牙周炎造模部位进行micro-CT成像分析,苏木精-伊红(H-E)染色观察组织病理学变化。结果与H组相比,牙周炎各阶段分组患者GCF中ErbB4的表达量显著升高,Nrg4的表达量显著降低(P<0.05)。在体外使用IFNγ+LPS刺激原代BMDMs引起其内ErbB4 mRNA表达显著上调(P<0.01)。使用Nrg4重组蛋白处理骨髓来源M1巨噬细胞后,其存活能力显著降低,细胞凋亡水平显著升高(P<0.01)。动物实验micro-CT成像分析表明,使用Nrg4重组蛋白治疗牙周炎大鼠模型显示出较好的疗效,与M组相比,T组釉-牙骨质界和牙槽骨嵴(CEJ-ABC)间的距离明显缩小,骨矿物质密度(BMD)和骨体积分数(BV/TV)明显升高(P<0.01);而与S组相比,T组上述4个参数的测定结果无明显变化(P>0.05)。结论GCF中ErbB4和Nrg4的表达水平能够作为潜在牙周炎患者炎症水平的标志物,Nrg4重组蛋白治疗牙周炎在在体动物模型水平上显示出了较好的疗效。
Objective To investigate the expression of neuregulin 4(Nrg4)and its receptor ErbB4 in the gingival crevicular fluid(GCF)of patients with periodontitis and their effects on periodontal tissue inflammation and alveolar bone loss in rats with experimental periodontitis.Methods A total of 80 healthy subjects and periodontitis patients were enrolled and divided into four groups(n=20):a health group(group H),a periodontitis group(group G),a stageⅡgrade B periodontitis group(group P1)and a stageⅢgrade B periodontitis group(group P2).Their GCF was collected,and the levels of ErbB4 and Nrg4 were detected by ELISA.Primary bone marrow-derived macrophages(BMDMs)and dendritic cells(BMDCs)were isolated and stimulated in vitro with IFNγ+LPS,and the intracellular expression of EGFR,ErbB2,ErbB3,and ErbB4 mRNA was measured by qPCR.Bone marrow-derived M1 macrophages were treated with Nrg4,and cell viability was detected by CCK-8 assay.Cell apoptosis were detected by flow cytometry with Annexin V/PI.An in vivo experimental periodontitis model of rats was constructed.A total of 15 rats were randomly divided into three groups(n=3):a sham operation group(group S),a model group(group M)and a Nrg4 recombinant protein treatment group(group T).The rats were subjected to micro-CT imaging analysis,followed by H-E staining and pathological analysis.Results Compared with group H,the expression of ErbB4 in the GCF of patients at each stage of periodontitis significantly increased(P<0.05),but the expression of Nrg4 significantly decreased(P<0.05).Meanwhile,in vitro stimulation of BMDMs with IFNγ+LPS resulted in significant up-regulation of ErbB4 mRNA expression(P<0.01).After Nrg4 treatment,the viability of bone marrow-derived M1 macrophages were significantly reduced,while cell apoptosis remarkably increased(P<0.01).According to micro-CT imaging analysis,Nrg4 recombinant protein was effective in the treatment of periodontitis.Compared with group M,group T showed remarkable decreases in the distance between the cement-enamel junction and the alveolar bone(CEJ-ABC),and increases in bone mineral density(BMD)and bone/tissue volume(BV/TV)(P<0.01).There was no significant difference as to the above four parameters between group T and group S.Conclusions The levels of ErbB4 and Nrg4 in GCF can be used as potential markers of inflammation in patients with periodontitis.Nrg4 recombinant protein shows good therapeutic effects in the treatment of periodontitis in vivo.
作者
张晓莉
马嘉平
ZHANG Xiaoli;MA Jiaping(Department of Stomatology,the People′s Hospital of Xinjiang Uygur Autonomous Region,Urumqi,Xinjiang 830001,China)
出处
《徐州医科大学学报》
CAS
2022年第5期318-325,共8页
Journal of Xuzhou Medical University
基金
新疆维吾尔自治区自然科学基金(2017D01C143)。
