Gardenia jasminoides Ellis is abundant in crocin and has a longstanding historical usage both as a dietary and natural ethnic medicine. Enhanced studies have increasingly revealed the intricate interplay between glycolipid metabolism and gut microbiota, wherein their imbalance is regarded as a pivotal indicator of metabolic disorders. Currently, the precise molecular mechanism of the crude extract of crocin from Gardenia jasminoides Ellis (GC) targeting gut microbiota to regulate glycolipid metabolism disorder is still unclear. Firstly, we explored the effect of GC on digestive enzymes (α-amylase and α-glucosidase) in vitro. Secondly, we investigated the effect of GC on the physical and chemical parameters of high-fat diet (HFD) rats, such as body weight change, fasting blood glucose and lipid levels, and liver oxidative stress and injury. Then, 16S rDNA sequencing was used to analyze the effects of GC on the composition and structure of gut microbiota. Finally, the impact of GC on the TLR4/Myd88/NF-κB signaling pathway in the intestine was assessed by Western Blotting. In the present study, GC was found to exhibit a hypoglycemic effect in vitro, by inhibition of digestive enzymes. In animal experiments, we observed that GC significantly reduced fasting blood glucose, TC, and TG levels while increasing HDL-C levels. Additionally, GC demonstrated hepatoprotective properties by enhancing liver antioxidative capacity through the upregulation of SOD, CAT, and GSH-Px, while reducing ROS. 16S rDNA sequencing results showed that GC had a significant effect on the gut microbiota of HFD rats, mainly by reducing the ratio of Firmicutes/Bateroidota, and significantly affected the genera related to glycolipid metabolism, such as Akkermansia, Ligilactobacillus, Lactobacillus, Bacteroides, Prevotellaceae, etc. The Western Blotting results demonstrated that GC effectively downregulated the protein expressions of TLR4, Myd88, and NF-κB in the intestine of HFD rats, indicating that GC could target the TLR4/Myd88/NF-κB pathway to interfere with glycolipid metabolism disorder. Correlation analysis revealed that GC could target the Akkermansia-TLR4/Myd88/NF-κB pathway axis which attenuates glycolipid metabolism disorder. Therefore, this study establishes the foundation for GC as a novel therapeutic agent for glycolipid metabolism disorder chemoprevention, and it introduces a novel methodology for harnessing the potential of natural botanical extracts in the prevention and treatment of metabolic syndrome.

The primary active compound in vine tea is dihydromyricetin (DMY), which has a longstanding history as a dietary supplement and traditional ethnic medicine. However, the precise molecular mechanism by which vine tea dihydromyricetin extract (VDMY) regulates glucolipid metabolic disorder remains unclear. In this study, we first assessed the effect of VDMY on various physiological parameters in db/db mice, followed by RNA sequencing (RNA-seq) to identify key signaling pathways affected by VDMY in liver tissues. We also examined the impact of VDMY on the liver’s TLR4/MyD88/NF-κB and FOXO1 pathways using Western blotting. Our results showed that VDMY significantly reduced fasting blood glucose (FBG), total cholesterol (TC), triglycerides (TGs), and low-density lipoprotein cholesterol (LDL-C), while increasing high-density lipoprotein cholesterol (HDL-C) levels. Additionally, VDMY enhanced the liver’s antioxidant capacity by upregulating superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH), while lowering malondialdehyde (MDA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), thus alleviating liver damage. RNA-seq analysis further revealed that VDMY influenced multiple biological processes, including transcription, glycolysis, gluconeogenesis, and redox reactions, suggesting that its effects may be mediated through the TLR4/MyD88/NF-κB and FOXO1 pathways. Additionally, Western blot analysis revealed that VDMY effectively downregulated the expressions of TLR4, MyD88, NF-κB, and FOXO1 proteins in the liver of db/db mice, indicating that VDMY could target these pathways to intervene glucolipid metabolism dysfunction.

The antioxidant N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD), widely used in tire rubber production, and its oxidative transformation product 6PPD-quinone (6PPD-Q), have become emerging urban environmental pollutants distributed worldwide, which can cause respiratory exposure in humans through atmospheric particulate matter and aerosols, potentially damaging the respiratory system; moreover, studies have suggested that such exposure is associated with the occurrence and development of lung cancer. In this study, a multidimensional computational research framework integrating network toxicology, weighted gene co-expression network analysis (WGCNA), machine learning, molecular docking, and molecular dynamics simulation was constructed to systematically decipher the dynamic molecular mechanisms by which 6PPD regulates lung cancer progression. Through cross-screening of multiple databases, 35 common interaction targets between 6PPD and lung cancer were identified, and candidate hub genes were screened out. GO and KEGG analyses showed that the targets were enriched in cancer-related pathways such as chemical carcinogenesis-reactive oxygen species and VEGF, suggesting that oxidative stress may be the core regulatory direction. The Ridge machine learning prediction model achieved an accuracy of 1.000 (AUC = 0.985) in the training set, and also exhibited good generalization ability in two independent validation datasets, and further screened out CA4, CDC25C, and NQO1 as core biomarkers; all three were significantly overexpressed in lung cancer tissues and exhibited good diagnostic efficacy for lung cancer. Molecular docking and 100 ns molecular dynamics simulation showed that 6PPD possessed strong spontaneous binding ability toward NOX4 (−8.0 kcal/mol), NQO1 (−7.9 kcal/mol), KDR, CA4, and CDC25C, and the complexes formed with the core targets remained generally stable throughout the simulations, suggesting that 6PPD can persistently bind to target proteins through stable intermolecular interactions. Trajectory hierarchical clustering analysis further showed that the conformational dynamic characteristics of each complex differed; among them, the KDR conformation was the most stable, whereas NQO1 and CA4 showed higher conformational diversity. In summary, the results of this study suggest that 6PPD may participate in oxidative stress pathway activation, DNA damage, cell cycle regulation, and tumor angiogenesis through multi-target synergistic effects, and be associated with lung cancer progression; this provides a computational pathway for deciphering the carcinogenic molecular mechanisms of environmental pollutants and assessing respiratory carcinogenic risk, and the core targets may provide candidate molecular markers for early warning and intervention of lung cancer.