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预防医学  2023, Vol. 35 Issue (11): 961-965    DOI: 10.19485/j.cnki.issn2096-5087.2023.11.010
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牙龈干细胞外泌体对牙周炎大鼠IL-6和TNF-α表达的影响
姜可1, 肖威1, 王权炳2, 陈虹1,3
1.蚌埠医学院口腔医学院,安徽 蚌埠 233030;
2.浙江省人民医院,浙江 杭州 310014;
3.浙江医院,浙江 杭州 310030
Effect of gingival stem cells-derived exosomes on interlukin (IL)-6 and tumor necrosis factor (TNF)-α expression in rats with periodontitis
JIANG Ke1, XIAO Wei1, WANG Quanbing2, CHEN Hong1,3
1. School of Stomatology, Bengbu Medical College, Bengbu, Anhui 233030, China;
2. Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang 310014, China;
3. Zhejiang Hospital, Hangzhou, Zhejiang 310030, China
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摘要 目的 探讨牙龈干细胞外泌体(GMSC-Exos)对牙周炎大鼠白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)表达的影响,为牙周炎治疗提供参考。方法 40只8周龄SPF大鼠随机分为空白组、牙周炎组、GMSC-Exos组和磷酸盐缓冲液(PBS)组,除空白组外的其他3组大鼠采用结扎丝法构建牙周炎模型,造模成功后空白组和牙周炎组大鼠不进行治疗,GMSC-Exos组和PBS组大鼠分别注射20 μL GMSC-Exos和PBS。4周后进行大鼠牙周探诊检查,采用酶联免疫吸附试验(ELISA)检测外周血TNF-α和IL-6浓度;采用实时荧光定量PCR技术检测TNF-α和IL-6相关基因;取左上颌牙周组织进行病理检查。比较各组大鼠牙周临床指标、IL-6和TNF-α水平。结果 4周后,GMSC-Exos组大鼠龈沟出血指数、牙龈指数、探诊深度和菌斑指数(1.87±0.41,1.03±0.19,1.91±0.09和1.11±0.17)高于空白组大鼠(0.96±0.31,0.83±0.31,1.09±0.05,1.01±0.38),但低于牙周炎组(2.65±0.50,1.36±0.22,2.61±0.07和1.51±0.26)和PBS组大鼠(2.44±0.50,1.23±0.20,2.49±0.10和1.39±0.28);GMSC-Exos组大鼠外周血IL-6和TNF-α浓度[(205.97±11.47)和(90.11±8.57)pg/mL]高于空白组大鼠[(143.10±4.87)和(80.07±5.13)pg/mL],低于牙周炎组[(367.33±13.89)和(158.29±13.10)pg/mL]和PBS组大鼠[(364.23±13.62)和(140.60±11.73)pg/mL];GMSC-Exos组大鼠牙龈组织IL-6和TNF-α mRNA表达量(1.09±0.14和1.61±0.29)高于空白组大鼠(0.99±0.10和1.06±0.14),低于牙周炎组(1.63±0.09和3.63±0.26)和PBS组大鼠(1.58±0.11和3.79±0.32),差异均有统计学意义(均P<0.05)。病理检查结果显示,与牙周炎组和PBS组比较,GMSC-Exos组大鼠牙周组织破坏程度、炎症细胞浸润程度及牙槽骨吸收程度减轻,结合上皮未见明显的根方增殖。结论 GMSC-Exos可降低牙周炎大鼠IL-6和TNF-α基因表达,减轻牙周炎症和牙槽骨吸收。
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姜可
肖威
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陈虹
关键词 牙龈干细胞外泌体牙周炎白细胞介素-6肿瘤坏死因子-α炎症因子    
AbstractObjective To evaluate the effect of gingival stem cells-derived exosomes (GMSC-Exos) treatment on the expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in rats with periodontitis, so as to provide the evidence for periodontitis treatment. Methods Forty specific pathogen-free (SPF) rats at ages of 8 weeks were randomly divided into 4 groups, including the blank group, periodontitis group, GMSC-Exos group and PBS group. Rats in the periodontitis group, GMSC-Exos group and PBS group were modeled for periodontitis using the ligature method. Rats in the blank group and periodontitis group were given no treatment, while rats in the GMSC-Exos group and PBS group were given 20 μL GMSC-Exos and PBS by injection, respectively. The periodontal index was measured in all rats 4 weeks post-treatment, and the TNF-α and IL-6 levels were measured in rat serum samples using enzyme-linked immunosorbent assay (ELISA). The TNF-α and IL-6 gene expression was quantified using the polymerase chain reaction (PCR) assay in the gingival tissues of the rat left upper maxillary area, and the periodontal tissues in the left upper maxillary areas were sampled for pathological examinations. Periodontal clinical indexes, IL-6 and TNF-α levels were compared in each group. Results The gingival sulcus bleeding index, gingival index, probing depth, and plaque index in the GMSC-Exos group (1.87±0.41, 1.03±0.19, 1.91±0.09 and 1.11±0.17) were higher than those in the blank group (0.96±0.31, 0.83±0.31, 1.09±0.05 and 1.01±0.38), but lower than those in the periodontitis group (2.65±0.50, 1.36±0.22, 2.61±0.07 and 1.51±0.26) and PBS group (2.44±0.50, 1.23±0.20, 2.49±0.10 and 1.39±0.28) (all P<0.05). The serum IL-6 and TNF-α levels in the GMSC-Exos group [(205.97±11.47) and (90.11±8.57) pg/mL] were higher than those in the blank group [(143.10±4.87) and (80.07±5.13) pg/mL], but lower than those in the periodontitis group [(367.33±13.89) and (158.29±13.10) pg/mL] and PBS group [(364.23±13.62) and (140.60±11.73) pg/mL] (all P<0.05). The IL-6 and TNF-α mRNA expression in the rat gingival tissues in the GMSC-Exos group (1.09±0.14 and 1.61±0.29) was higher than that in the blank group (0.99±0.10 and 1.06±0.14), but lower than that in the periodontitis group (1.63±0.09 and 3.63±0.26) and PBS group (1.58±0.11 and 3.79±0.32) (all P<0.05). Pathological examinations showed alleviation of periodontal tissue destruction, inflammatory cell infiltration and alveolar bone resorption, and no obvious root dental root regeneration in the junctional combined epithelium in the GMSC-Exos group relative to the periodontitis group and the PBS group. Conclusion Administration of GMSC-Exos may reduce periodontal inflammation and alveolar bone resorption by inhibiting IL-6 and TNF-α expression in rats with periodontitis.
Key wordsgingival stem cell    exosome    periodontitis    interleukin-6    tumor necrosis factor-α    inflammatory factor
收稿日期: 2023-07-26      修回日期: 2023-10-04      出版日期: 2023-11-10
中图分类号:  R781.4  
基金资助:浙江省医药卫生科技计划项目(2020KY449); 蚌埠医学院研究生科研创新项目(Byycx22147)
通信作者: 陈虹,E-mail:hssy81@126.com   
作者简介: 姜可,硕士研究生在读
引用本文:   
姜可, 肖威, 王权炳, 陈虹. 牙龈干细胞外泌体对牙周炎大鼠IL-6和TNF-α表达的影响[J]. 预防医学, 2023, 35(11): 961-965.
JIANG Ke, XIAO Wei, WANG Quanbing, CHEN Hong. Effect of gingival stem cells-derived exosomes on interlukin (IL)-6 and tumor necrosis factor (TNF)-α expression in rats with periodontitis. Preventive Medicine, 2023, 35(11): 961-965.
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http://www.zjyfyxzz.com/CN/10.19485/j.cnki.issn2096-5087.2023.11.010      或      http://www.zjyfyxzz.com/CN/Y2023/V35/I11/961
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