黄芩苷对呼吸道合胞病毒感染的Hep-2细胞凋亡和炎性因子表达的影响

Effects of baicalin on apoptosis and expression of inflammatory factors in Hep-2 cells infected by respiratory syncytial virus

目的探讨黄芩苷对呼吸道合胞病毒感染的Hep-2细胞凋亡和炎性因子表达的影响.方法使用呼吸道合胞病毒处理Hep-2细胞,记为感染组,以未经呼吸道合胞病毒处理Hep-2细胞作为对照组.采用黄芩苷浓度为12.5、25、50μmol/L处理感染的Hep-2细胞,记为低剂量组、中剂量组、高剂量组.检测细胞增殖和细胞凋亡;蛋白免疫印迹法(Western blot)检测B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved-cas-3)、磷酸化的核因子-κB(p-NF-κB)、核因子-κB(NF-κB)、p-p38、p38蛋白表达;酶联免疫吸附法检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、IL-1β表达.结果与对照组相比,感染组细胞活性、Bcl-2蛋白表达降低(P<0.05),凋亡率、Cleaved-cas-3、Bax、p-NFκB、p-p38蛋白表达增加(P<0.05),TNF-α、IL-6、IL-1β表达增加(P<0.05).与感染组相比,低剂量组、中剂量组、高剂量组细胞活性、Bcl-2蛋白表达增加(P<0.05),凋亡率、Cleaved-cas-3、Bax、p-NFκB、p-p38蛋白表达降低(P<0.05),TNF-α、IL-6、IL-1β表达降低(P<0.05).结论黄芩苷可能通过降低NF-κB、p38信号通路,减轻呼吸道合胞病毒感染的Hep-2细胞凋亡和炎性因子表达.

OBJECTIVE To investigate the effects of baicalin on the apoptosis and expression of inflammatory factors in Hep-2 cells infected with respiratory syncytial virus.METHODS Hep-2 cells were treated with respiratory syncytial virus and recorded as the infected group,while Hep-2 cells without respiratory syncytial virus treatment were used as the control group.Infected Hep-2 cells were treated with baicalin concentrations of 12.5,25,and 50μmol/L and recorded as low-dose group,middle-dose group,and high-dose group,respectively.MTT method was used to detect cell proliferation and flow cytometry was used to detect cell apoptosis;B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax),activated cysteine-containing aspartate proteolytic enzyme 3(Cleavedcas-3),phosphorylated nuclear factor-kB(p-NF-kB),nuclear factor-kB(NF-kB),P-p38,and p38 protein expression were detected by Western Blotting;the expressions of tumor necrosis factor-α(TNF-α),interleukin(IL)-6 and IL-1βwere detected by enzyme-linked immunosorbent assay.RESULTS Compared with the control group,the cell activity and Bcl-2 protein expression in the infection group decreased(P<0.05),while the apoptosis rate,cleaved-cas-3,Bax,p-NF kB,P-p38 protein expression increased(P<0.05)as well as TNF-α,IL-6,IL-1βexpression(P<0.05).Compared with the infection group,the cell activity and Bcl-2 protein expression increased(P<0.05)in the low-dose,middle-dose and high-dose groups,while the apoptosis rate,cleaved-cas-3,Bax,p-NFkB,and p-p38 protein expression decreased(P<0.05)as well as TNF-α,IL-6,IL-1βexpression(P<0.05).CONCLUSION Baicalin might attenuated the apoptosis and inflammatory factors expression in respiratory syncytial virus-infected Hep-2 cells byreducing theNFkB and p38 signaling pathways.