摘要
目的利用牙周膜干细胞研究细胞因子IL38是否参与牙周炎的炎症反应过程。方法利用20μg/mL牙龈卟啉单胞菌脂多糖Pg-LPS刺激牙周膜干细胞24 h建立牙周炎炎症体外模型;利用IL38 siRNA、IL38过表达质粒及相应对照进行牙周膜干细胞的转染,进而研究IL38对牙周膜干细胞炎症反应的影响;利用qPCR方法检测牙周膜干细胞中炎症细胞因子的释放,利用Western blot检测P50及P65蛋白质表达情况,利用CCK-8实验检测牙周膜干细胞的增殖情况。结果IL36亚家族中只有IL38在Pg-LPS刺激牙周膜干细胞中显著上升;在正常及炎症状态下,IL38沉默降低了细胞因子IL17A、TNFA及IL1B的mRNA表达水平,但IL38过表达逆转了上述结果;Western blot结果表明,IL38过表达促进P50及P65蛋白质表达水平;IL38过表达也增加了牙周膜干细胞的增殖,而IL38沉默抑制了其增殖能力。结论在体外实验中,IL38具有刺激牙周膜干细胞炎症细胞因子表达及增殖的能力。
Objective Our purpose is to investigate whether IL38 participates in the inflammatory responses of periodontitis in vitro.Methods In our work,periodontal inflammation cell model was established with 20μg/ml Pg-LPS(Porphyromonas gingivalis lipopolysaccharide)for 24 h.IL38 siRNA,IL38 over-expressed vector and corresponding controls were transfected into human periodontal ligament stem cells(HPDLSCs)to explore how IL38 functions in periodontal inflammation.Cytokine mRNA expression was examined by qPCR and protein expression was measured by western blot.We used CCK-8 assay to examine cell proliferation.Results The results discovered that IL38,the only member in IL36 subfamily,was significantly increased in HPDLSCs after Pg-LPS stimulation.More importantly,IL38 silencing decreased IL17A,TNFA and IL1B under physiological and inflammatory conditions while the cytokines above were reduced when IL38 is over-expressed in HPDLSCs,which accompanied with an enhanced P50 and P65 protein expression in HPDLSCs after Pg-LPS treatment.Moreover,our results also found IL38 over-expression increased HPDLSC proliferation while IL38 silencing inhibited its proliferation.Conclusion Our data demonstrated IL38 increased inflammatory cytokine production and proliferation of HPDLSCs in vitro.
作者
徐曦
郎广平
吴梦鑫
韩盈盈
Xu Xi;Lang Guangping;Wu Mengxin;Han Yingying(Special Key Laboratory of Oral Diseases Research,Hospital of Stomatology/School of Stomatology,Zunyi Medical University,Zunyi Guizhou 563099,China;Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education,Zunyi Medical University,Zunyi Guizhou 563099,China)
出处
《遵义医科大学学报》
2023年第8期738-745,共8页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:82160181)
贵州省卫生健康委科学技术基金资助项目(NO:2023-201)
遵义市科技联合基金[NO:遵市科合HZ字(2022)377、(2021)284]。
关键词
IL38
牙周炎
牙周膜干细胞
炎症
增殖
IL38
periodontitis
human periodontal ligament stem cells
inflammation
proliferation