与脐带间充质干细胞共培养对脂多糖诱导的牙髓细胞炎症的影响

Effect of co-culture with umbilical cord mesenchymal stem cells on lipopolysaccharide-induced inflammation in dental pulp cells

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.