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

Area of Science:

• Pharmacology
• Biochemistry
• Traditional Chinese Medicine

Background:

• Nonalcoholic steatohepatitis (NASH) is a growing health concern with limited treatment options.
• Erhuangquzhi granules (EQG) show clinical efficacy in NASH, but active components and mechanisms are unclear.

Purpose of the Study:

• To identify active components in EQG that target tumor necrosis factor alpha (TNF-α) for NASH treatment.
• To elucidate the molecular mechanisms underlying EQG's therapeutic effects in NASH.

Main Methods:

• Utilized a surface plasmon resonance (SPR) biosensor-based active ingredient recognition system (SPR-AIRS).
• Immobilized recombinant TNF-α protein on an SPR chip for ligand screening.
• Employed UPLC‒MS/MS for identification and SPR for affinity constant determination of active compounds.
• Assessed the inhibitory effects of identified compounds on TNF-α-induced apoptosis and inflammatory pathways in cell models.

Main Results:

• Identified nuciferine (NF), lirinidine (ID), and O-nornuciferine (NNF) from lotus leaves as TNF-α ligands.
• Determined affinity constants for NF, ID, and NNF binding to TNF-α.
• Demonstrated that NF, ID, and NNF inhibit TNF-α-induced apoptosis and reduce key inflammatory markers (IL-6, IL-1β) and signaling pathways (IKKβ, IκB phosphorylation) in relevant cell lines.

Conclusions:

• Discovered a class of novel small-molecule TNF-α inhibitors from EQG.
• Provides insights into the material basis and molecular mechanisms of EQG for NASH treatment.
• Highlights the potential of NF, ID, and NNF as therapeutic agents for NASH.