摘要
目的探讨解毒祛瘀滋阴方冻干粉与含药血清对脂多糖(LPS)诱导的小鼠单核巨噬细胞RAW264.7炎症信号通路的影响。方法体外培养RAW264.7细胞,随机分为对照组、LPS组、解毒祛瘀滋阴方含药血清组、冻干粉组、LPS加含药血清组及LPS加冻干粉组。干预24 h后,采用CCK-8法筛选含药血清和冻干粉的最佳浓度并检测二者对细胞活力的影响;采用ELISA法测定细胞上清液中肿瘤坏死因子-α(TNF-α)含量;采用q RT-PCR法检测核转录因子-κB(NF-κB)、TNF-αm RNA的表达;采用Western blotting法检测NF-κB蛋白表达水平;采用LC-MS检测解毒祛瘀滋阴方含药血清中的有效成分。结果与对照组比较,LPS刺激组TNF-α炎性因子水平、NF-κB、TNF-αm RNA表达水平和NF-κB蛋白表达水平显著升高(P<0.05),含药血清组TNF-α炎性因子水平、NF-κB、TNF-αm RNA表达水平和NF-κB蛋白表达水平较冻干粉组降低明显(P<0.05);与LPS组比较,LPS加含药血清组TNF-α炎性因子水平、NF-κB、TNF-αm RNA表达水平和NF-κB蛋白表达水平降低较LPS加冻干粉组明显(P<0.05);解毒祛瘀滋阴方含药血清中检测到芍药苷和阿魏酸。结论解毒祛瘀滋阴方冻干粉和含药血清均具有抑制炎症信号通路的作用。
Objective To study the effects of Jiedu Quyu Ziyin Prescription（JQZP）-treated freeze dried powder and drug-containing serum on the inflammatory signal pathway of monocyte-macrophage induced by LPS（lipopolysaccharide） in mice. MethodsMonocyte-macrophage cells were cultured in vitro and randomly divided into blank group, LPS stimulation group, drug-containing serum group, freeze dried powder group, LPS ＋ drug-containing serum group, and LPS ＋ freeze dried powder group. After 24 h intervention, the optimal concentrations of drug-containing serum and freeze dried powder were screened by CCK8 method and the cell viability was measured respectively. The content of tumor necrosis factor（TNF-α） in cell serum was measured by ELISA. Real-time PCR was employed to test the expression of TNF-α m RNA and nuclear transcription factor kappa-light-chain-enhancer of activated B cells（NF-κB）. Western-blot was used to detect the expression of NF-κB protein. The LC-MS was used to detect the active ingredients in the drug-containing serum. Results Compared with the blank group, the expression of TNF-α level, NF-κB, TNF-α m RNA and NF-κB protein in LPS stimulation group were significantly increased（P 0.05）. Compared with the LPS stimulation group, the TNF-α level, NF-κB, TNF-α m RNA and the expression of NF-κB protein in the LPS plus serum group were significantly lower than those in the LPS plus freeze-dried powder group（P 0.05）. Paeoniflorin and ferulic acid were detected in the drug-containing serum. Conclusion JQZP freeze-dried powder and drug-containing serum all have the effect of inhibiting the inflammatory signaling pathway.
作者
嵇丽娜
刘丹
邓贤
李荣群
JI Li-na;LIU Dan;DENG Xian;LI Rong-qun(Zhej iang Chinese Medical University,Hangzhou 310053,China)
出处
《中草药》
CAS
CSCD
北大核心
2018年第16期3868-3874,共7页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金面上项目(81673863)
浙江省自然科学基金资助项目(LY15H270007)
关键词
解毒祛瘀滋阴方
冻干粉
含药血清
炎症信号通路
芍药苷
阿魏酸
Jiedu Quyu Ziyin Prescription
freeze dried powder
drug-containing serum
inflammatory signal pathway
paeoniflorin
ferulic acid
