白头翁总皂苷对人轮状病毒NSP4致细胞炎性因子水平的影响

Effects of Pulsatilla Saponin on the levels of cytoinflammatory factors induced by human rotavirus NSP4

目的:探讨白头翁总皂苷对人轮状病毒(RV)非结构蛋白4(NSP4)引起的细胞炎性因子水平及核因子κB(NF-κB) p65蛋白表达的影响。方法:用人RV NSP4真核表达质粒转染HT-29细胞,使用不同浓度的白头翁总皂苷干预HT-29-NSP4细胞。实时定量PCR检测NSP4 mRNA表达,酶联免疫吸附测定(ELISA)检测细胞培养上清液中炎性因子白细胞介素6(IL-6)、IL-8水平,免疫印迹法检测NF-κB p65蛋白表达。结果:实时定量PCR显示,HT-29-NSP4组NSP4 mRNA表达量较HT-29-NC组显著升高(P<0.01)。与HT-29-NC组比较,HT-29-NSP4组细胞上清液中IL-6及IL-8水平均显著升高,细胞NF-κB p65蛋白相对表达量升高(均P<0.05)。与HT-29-NSP4组比较,白头翁总皂苷处理后细胞上清液IL-6及IL-8水平均显著降低,细胞NF-κB p65蛋白相对表达量降低(均P<0.05)。结论:白头翁总皂苷可能通过作用于NF-κB通路抑制人RV NSP4引起的细胞炎性因子释放。

Objective:To investigate the effect of Pulsatilla Saponin on the expression of inflammatory factors and NF-κB p65 protein induced by human rotavirus NSP4.Methods:Human HT-29 cells were transfected with human RV NSP4 eukaryotic expression plasmid and then were treated with varies concentrations of Pulsatilla Saponins.After treatment of HT-29-NSP4 cells with different concentrations of Pulsatilla saponins solution,the culture supernatant were collected to detected the expression of IL-6 and IL-8 by ELISA respectively,and cell pellets were collected to determine the expression of NF-κB p65 protein by Western Blot.Results:The expression of NSP4 mRNA in HT-29-NSP4 group was significantly increased compared with HT-29-NC group(P<0.01).Compared with HT-29-NC group,the concentrations of IL-6 and IL-8 levels were significantly increased in HT-29-NSP4 group,and the expression of NF-κB p65 protein in HT-29-NSP4 group was significant elevated(all P<0.05).Compared with HT-29-NSP4 group,the expression levels of IL-6,IL-8 and NF-κB p65 protein in Pulsatilla saponins treatment group were decreased(all P<0.05).Conclusion:Pulsatilla saponins may inhibit the release of cellular inflammatory factors induced by human RV NSP4 by acting on the NF-κB pathway.