1. Academic Validation
  2. Biosensor-based active ingredient recognition system for screening TNF-α inhibitors from lotus leaves

Biosensor-based active ingredient recognition system for screening TNF-α inhibitors from lotus leaves

  • Anal Bioanal Chem. 2023 Jan 31. doi: 10.1007/s00216-023-04565-2.
Jia-Wei Guan # 1 Yu Xu # 1 Wei Yu 1 Li-Hua Wei 1 Rui Xue 1 Hang Yu 1 Jing-Min Fan 1 Hong-Zheng Wang 1 Abudumijiti Abulizi 1 Wen Chen 1 Bo Han 2
Affiliations

Affiliations

  • 1 School of Pharmacy/Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, Shihezi University, Shihezi, 832002, People's Republic of China.
  • 2 School of Pharmacy/Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, Shihezi University, Shihezi, 832002, People's Republic of China. 524683221@qq.com.
  • # Contributed equally.
Abstract

Erhuangquzhi granules (EQG) have been clinically proven to be effective in nonalcoholic steatohepatitis (NASH) treatment. However, the active components and molecular mechanisms remain unknown. This study aimed to screen active components targeting tumor necrosis factor α (TNF-α) in EQG for the treatment of NASH by a surface plasmon resonance (SPR) biosensor-based active ingredient recognition system (SPR-AIRS). The amine-coupling method was used to immobilize recombinant TNF-α protein on an SPR chip, the specificity of the TNF-α-immobilized chip was validated, and nine medicinal herbs in EQG were prescreened. Nuciferine (NF), lirinidine (ID), and O-nornuciferine (NNF) from lotus leaves were found and identified as TNF-α ligands by UPLC‒MS/MS, and the affinity constants of NF, ID, and NNF to TNF-α were determined by SPR experiments (Kd = 61.19, 31.02, and 20.71 µM, respectively). NF, ID, and NNF inhibited TNF-α-induced Apoptosis in L929 cells, the levels of secreted IL-6 and IL-1β were reduced, and the phosphorylation of IKKβ and IκB was inhibited in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In conclusion, a class of new active small-molecule TNF-α inhibitors was discovered, which also provides a valuable reference for the material basis and mechanism of EQG action in NASH treatment.

Keywords

Lirinidine; Lotus leaf; Nuciferine; O-Nornuciferine; SPR-UPLC‒MS/MS; TNF-α inhibitor.

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