摘要
目的探讨祛风扶正散调控Toll样受体4(TLR4)/核转录因子κB(NF-κB)信号通路治疗周围性面瘫的作用机制。方法建立RAW264.7细胞炎症模型,用不同浓度祛风扶正散受试物处理炎症模型细胞,采用Griess reagent法检测细胞一氧化氮(NO)的分泌量,酶联免疫吸附试验(ELISA)法检测细胞上清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、前列腺素2(PEG 2)的水平,活性氧荧光探针(DCFH-DA)法检测细胞内活性氧(ROS)的水平,实时荧光定量PCR(RT-PCR)检测细胞TNF-α、IL-1βmRNA的表达,蛋白印迹法(Western blot)检测细胞p65、磷酸化p65(p-p65)、IκBα、磷酸化IκBα(p-IκBα)蛋白表达。结果祛风扶正散可显著降低炎症模型细胞NO、ROS、TNF-α、IL-1β、IL-6及PEG 2表达水平(F=338.731~5790.712,P<0.01),下调TNF-α、IL-1βmRNA表达(F=134.430、168.217,P<0.01),抑制p-p65和p-IκBα蛋白表达、IκBα降解及p65核易位(F=17.793~97.613,P<0.01)。结论祛风扶正散治疗周围性面瘫的作用机制可能与抑制本病相关TLR4/NF-κB信号通路的活化及其下游促炎性细胞因子、炎症递质的释放有关。
Objective To investigate the mechanism of action of Qufeng Fuzheng powder in the treatment of peripheral facial paralysis by regulating the Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)signaling pathway.Methods RAW264.7 cells were used to establish an inflammation model,and then the cells were treated with different concentrations of Qufeng Fuzheng powder.The Griess reagent method was used to measure the secretion of nitric oxide(NO);ELISA was used to measure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),and prostaglandin-2(PEG 2)in cell supernatant;the DCFH-DA fluorescent probe was used to measure the level of reactive oxygen species(ROS)in cells;quantitative real-time PCR was used to measure the mRNA expression levels of TNF-αand IL-1β,and Western blot was used to measure the protein expression levels of p65,phosphorylated p65(p-p65),nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor,alpha(IκBα),and phosphorylated IκBα(p-IκBα).Results Qufeng Fuzheng powder significantly reduced the levels of NO,ROS,TNF-α,IL-1β,IL-6,and PEG 2 in cells(F=338.731-5790.712,P<0.01)and downregulated the mRNA expression levels of TNF-αand IL-1β(F=134.430,168.217;P<0.01).Moreover,it also inhibited the protein expression of p-p65 and p-IκBα,the degradation of IκBα,and the nuclear translocation of p65(F=17.793-97.613,P<0.01).Conclusion Qufeng Fuzheng powder exerts a therapeutic effect on peripheral facial paralysis possibly by inhibiting activation of the TLR4/NF-κB signaling pathway and the release of its downstream proinflammatory cytokines and inflammatory mediators.
作者
周丽华
周长凯
商秀玲
荆凡波
ZHOU Lihua;ZHOU Changkai;SHANG Xiuling;JING Fanbo(College of Pharmacy,Qingdao University,Qingdao 266071,China)
出处
《青岛大学学报（医学版）》
CAS
2023年第1期73-78,共6页
Journal of Qingdao University(Medical Sciences)
基金
山东省重大科技创新工程(2018CXGC1304)
山东省中医药科技发展项目(2019-0400、2021Q097)。
关键词
面神经麻痹
中药
祛风扶正散
TOLL样受体4
NF-ΚB
facial paralysis
traditional Chinese drugs
Qufeng Fuzhegn powder
Toll-like receptor 4
NF-kappa B
