摘要
目的:探讨人牙髓细胞(h DPCs)和人脐带间充质干细胞(hUCMSCs)共培养系统对脂多糖(LPS)诱导的炎症微环境的调节作用。方法:用不同浓度LPS(1μg/m L、10μg/m L、20μg/m L、50μg/m L)处理hDPCs和hUCMSCs,以获得诱导细胞炎症反应的适宜浓度。按1∶1的比例建立hDPCs与hUCMSCs直接及间接共培养体系,MTT法检测细胞增殖,分别采用qRT-PCR和酶联免疫吸附试验(ELISA)法检测IL-6、IL-10和HGF mRNA相对表达量和蛋白分泌量。结果:10μg/m L LPS可以诱导细胞炎症反应。与单培养hDPCs组相比,直接共培养系统可促进细胞增殖,提高IL-10和HGF mRNA表达量及蛋白分泌量,降低IL-6 m RNA表达量及蛋白分泌量(P<0.05)。在间接共培养系统中,IL-6、IL-10和HGF的蛋白分泌量也显示出相同的趋势。此外,与单培养组的hDPCs相比,来自共培养系统的h DPCs高表达IL-10和HGF mRNA,但低表达IL-6(P<0.05)。结论:与单培养h DPCs组相比,共培养系统在下调LPS诱导的牙髓细胞炎症方面表现出更强的能力。
Objective: To investigate the regulation effect of the co-culture of human dental pulp cells(hDPCs) and human umbilical cord mesenchymal stem cells(hUCMSCs) onlipopolysaccharide(LPS)-induced inflammatory microenvironment. Methods: To get suitable concentration of LPS for simulating inflammatory responses, hDPCs and hUCMSCs were cultured with different concentrations of LPS(1, 10, 20 and 50μg/m L). Then, direct and indirect co-culture system of hUCMSCs and hDPCs at ratio of 1∶1 were established in vitro with LPS of 10 μg/m L. Cell proliferation was detected by MTT assay. The mRNA and protein expression levels of interleukin(IL)-6, IL-10 and hepatocyte growth factor(HGF) were analyzed by quantitative reverse-transcription polymerase chain reaction(qRT-PCR) and enzyme-linked immunosorbent assay(ELISA), respectively. Results:LPS of 10 μg/m L could induce cell inflammation. The direct co-culture system enhanced cell proliferation, increased mRNA and protein levels of IL-10 and HGF, while decreased the level of IL-6 compared with hDPCsmonocultured with LPS(P<0.05). In indirect co-culture system, the protein levels of IL-6, IL-10 and HGF showed the same trend. Additionally, hDPCs from the co-culture system expressed extremely higher mRNA levels of IL-10 and HGF,but lower level of IL-6 compared with hDPCs monocultured with LPS(P<0.05). Conclusion: Co-culture with hUCMSCs might attenuate LPS-induced dental pulp cell inflammation within a certain range in vitro.
作者
何凯丽
刘玉莹
林田
陆彦玲
吴煜
Kaili He;Yuying Liu;Tian Lin;Yanling Lu;Yu Wu(Guangxi Medical University,Nanning 530021,China;Department of Pediatric Dentistry,Stomatology Hospital,Guangxi Medical University,Nanning 530021,China;Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction,Nanning 530021,China;Guangxi Key Laboratory of Oral and Maxillofacial Surgery Disease Treatment,Nanning 530021,China;Guangxi Clinical Research Center for Craniofacial Deformity,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2021年第2期217-228,共12页
Journal of Guangxi Medical University
基金
supported by the National Natural Science Foundation of China (No. 81660182
No. 82060200)。
关键词
脐带间充质干细胞
牙髓细胞
共培养
脂多糖
炎症因子
umbilical cord mesenchymal stem cells
dental pulp cells
co-culture
lipopolysaccharide
cytokines