The study aimed to optimize the processing conditions of Gardeniae Fructus with ginger juice (GFPG) and confirm its therapeutic effects and pharmacological mechanisms on cholestatic liver injury. Processing conditions were optimized using response surface methodology (RSM) and thermal analysis, focusing on geniposide content as a key active compound. Variables included processing time, moistening time, and the solid-liquid ratio. Optimal conditions were: ginger juice to Gardeniae Fructus ratio of 8:1 (w/v), processing temperature of 208 °C, moistening time of 3 hours, and processing time of 5 minutes. Pharmacological mechanisms were analyzed through network pharmacology, molecular docking, and experimental validation using alpha-naphthyl isothiocyanate (ANIT)-induced cholestatic liver injury in mice and lipopolysaccharide (LPS)-stimulated RAW264.7 cell models. In vivo, GFPG extract alleviated ANIT-induced cholestatic liver injury by improving liver function markers (AST, ALT, TBA, TBIL, DBIL) and modulating TLR4/NF-κB, FXR/PPAR-α, and PI3K/AKT/GSK-3β pathways. In vitro, it reduced LPS-induced production of inflammatory mediators (NO, TNF-α, IL-6, IL-1β) through TLR4/NF-κB pathway inhibition. This study established optimal processing methods for GFPG using RSM and thermal analysis, providing robust quantitative parameters. GFPG demonstrated significant therapeutic effects in cholestatic liver injury models, indicating its potential as a candidate for developing treatments for cholestatic hepatitis.

Background: Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by the deposition of senile plaques, neurofibrillary tangles, and neuronal dysfunction, resulting in severe cognitive and memory decline. The root of the Scrophulariaceae plant Rehmannia glutinosa (Gaertn.) DC. (Rehmanniae radix; RR) and its product Rehmanniae radix praeparata (RRP) possess high nutritional and medicinal value. Both show therapeutic potential for AD in traditional medical settings. However, the differences in their bioactive components and the mechanisms of action underlying their anti-AD effects remain unclear.Methods: In this study, APP/PS1 mice were used as the animal model of AD. Ultra-high-performance liquid chromatography coupled with Q-Exactive tandem mass spectrometry (MS/MS) (UPLC–QE-MS/MS), network pharmacology, proteomics, molecular docking, and 16S rRNA sequencing were used to investigate the differences in the medicinal components of RR and RRP and their mechanisms of action in the treatment of AD. The mechanisms of action of two identified critical components, ajugol and isoacteoside, were further verified in the D-galactose/AlCl₃-induced Institute of Cancer Research (ICR) mouse model of AD—with cognitive function evaluated using the Morris water maze and open-field tests—and the amyloid-beta (Aβ)-induced BV2 cell model of inflammation.Results: Ajugol and isoacteoside were identified as the key anti-AD bioactive compounds in RR and RRP, respectively, through UPLC–QE-MS/MS. Integrated network pharmacology, proteomics, and 16S rRNA sequencing implicated neuroinflammation, apoptosis, and autophagy as critical pathways for their anti-AD effects. Subsequently, in vivo and in vitro experiments demonstrated that ajugol exerted its effects mainly by modulating the TLR/NF-κB/NLRP3 and BCL-2/BAX/cytochrome C/caspase-3 pathways, while isoacteoside primarily acted via the LC3-Ⅱ/P62/p-mTOR/mTOR pathway. Ajugol and isoacteoside mitigated cognitive impairment in AD models, decreased Aβ plaque accumulation in hippocampal tissues, and attenuated inflammatory injury-induced cytotoxicity in BV2 microglia, thereby suppressing AD progression.Conclusion: In this work, we systematically elucidated the differential mechanisms underlying the anti-AD effects of ajugol and isoacteoside. We found that ajugol primarily acts via the TLR/NF-κB/NLRP3 and BCL-2/BAX/cytochrome C/caspase-3 pathways, while isoacteoside acts via the LC3-II/P62/p-mTOR/mTOR pathway. These findings establish a foundation for developing RRP-based complementary medicines and functional foods.

Due to the impact of various environmental factors, in conjunction with the ecological adaptability and biochemical diversity of Chinese medicinal materials, Codonopsis Radix from Shanxi Province has developed into two distinct commercial specifications, namely Tai Codonopsis Radix (TCP) and Lu Codonopsis Radix (LCP). The present study examined the relationships between TCP and LCP from Shanxi Province regarding their environment, secondary metabolites, and Traditional Chinese Medicinal plant authenticity using metagenomic, metabolomic, and comprehensive quality assessment methods. The findings revealed that the two types of CPs met the quality standards. With respect to the growing environment, TCP and LCP grow in the same longitudinal extent, with a maximum longitudinal span of only 53ʹ. Both regions presented similar natural environments conducive to the growth of CP. Additionally, the Lobetyolin content in TCP was greater than that in LCP. The soil microbial abundance in the TCP group was considerably significantly greater than that in the LCP group, and a new genus, Ascobolus, was found in TCP. Additionally, comparison of the metallic element data of the two groups soils revealed that the contents of Ti, Mg, Cr (P < 0.01) in the TCP group were significantly greater than those in the LCP group. Through correlation analysis, the ternary network of microbial-metal-metabolite interactions was constructed. The rhizosphere fungus Ascobolus was found to enhance the bioavailability of Zn via mineralization processes, thereby underscoring the significance of rhizosphere microecology in promoting plant growth. This study offers novel insights and approaches for the cultivation and processing of Codonopsis Radix.