title: Identification of NF-κB inhibitors following Shenfu injection and bioactivity-integrated UPLC/Q-TOF-MS and screening for related anti-inflammatory targets in vitro and in silico.
abstract: BACKGROUND
  Shenfu injection (SFI) is a commercial medicinal product approved by the China Food and Drug Administration that is widely used in the treatment of stroke and coronary heart disease. However, the material basis and the mechanism of SFI are not fully understood.
  
  
  OBJECTIVE
  With network pharmacology analysis, our research committed to identify the anti-inflammatory ingredients and mechanism of SFI by combining high-throughput screening.
  
  
  METHODS
  We developed a bioactivity-based UPLC/Q-TOF-MS method followed by network pharmacology and identified the anti-inflammatory active ingredients of SFI from two different perspectives of network computing and high throughput screening. Then we verified the anti-inflammatory effect of SFI in vitro with endothelial cells. After detecting the cell viability, the expression of interleukin-6 (IL-6), inhibitor of nuclear factor kappa-B kinase (IKK), phosphorylated IKK, phosphorylated NF-κB and phosphorylated IκB-α from the supernatant were determined.
  
  
  RESULTS
  SFI could significantly suppress inflammatory responses, and the mechanism may be via an NF-κB-dependent pathway. The results of high throughput screening (HTS) revealed that protopanaxadiol glycosides (ginsenosides Rb1, Rb2, Rb3, Rc and Rd), protopanaxatriol glycosides (ginsenosides Rg1, Rg2, Re, Rf and F1), diester-type alkaloids (fuziline and neoline) and aconine derivatives (mesaconine and benzoyl-mesaconine) have anti-NF-κB activity. The three compounds (including benzoyl-mesaconine, fuziline and neoline) are the first reported SFI compounds to have NF-κB inhibitor activity.
  
  
  CONCLUSIONS
  SFI may play a critical role in counteracting inflammation through the NF-κB signaling pathway. The active ingredients are protopanaxadiol glycosides, protopanaxatriol glycosides, diester-type alkaloids and aconine derivatives.
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Pan
authors: P
authors: Li
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Bin
authors: B
authors: Lv
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Xiaoqing
authors: X
authors: Jiang
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Ting
authors: T
authors: Wang
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Xianghui
authors: X
authors: Ma
authors: College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Nianwei
authors: N
authors: Chang
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China; College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China. Electronic address: wxy@tjutcm.edu.cn.
authors: Xiaoying
authors: X
authors: Wang
authors: State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.
authors: Xiumei
authors: X
authors: Gao
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002853:Chromatography, Liquid; D004365:Drugs, Chinese Herbal; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D013058:Mass Spectrometry; D016328:NF-kappa B / Q000037:antagonists & inhibitors*
chemical_list: D000893:Anti-Inflammatory Agents; D004365:Drugs, Chinese Herbal; D016328:NF-kappa B; C492793:Shen-Fu
keywords: Anti-inflammation; Benzoyl-mesaconine (Pubchem CID: 78358527); Fuziline (Pubchem CID: 157773); Ginsenoside Rb1 (Pubchem CID: 9898279); Ginsenoside Rc (Pubchem CID: 12855889); Ginsenoside Rd (Pubchem CID: 24721561); Ginsenoside Re (Pubchem CID: 441921); Ginsenoside Rf (Pubchem CID: 441922); Ginsenoside Rg1 (Pubchem CID: 441923); High throughput screening; Mesaconine (Pubchem CID: 54760335); NF- κB; Neoline (Pubchem CID: 120682); Network pharmacology; Shenfu injection