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Atherosclerosis is caused by multiple factors and has a complex pathogenesis. This study employed network pharmacology to identify key active constituents of Ilex kudingcha and their molecular targets related to atherosclerosis. Six active components were discovered, which interacted with 98 shared target genes. GO enrichment analysis revealed their roles in various biological functions, including transcription factors and cytokine activity, influencing processes such as wound healing and oxidative stress response. KEGG analysis highlighted involvement in pathways such as lipid and atherosclerosis, AGE-RAGE in diabetes and TNF signaling. Molecular docking showed good affinity of quercetin, kaempferol, and beta-sitosterol with key proteins such as AKT1 and VEGFA. These components exhibit anti-atherosclerosis effects through multitarget and multipathway mechanisms, with quercetin and kaempferol being potential key agents.

The quantitative analysis of multiple components with a single marker (QAMS) method was firstly established for simultaneous determination of 18 active components in Ilex kudingcha C. J. Tseng by HPLC. Using rutin, isochlorogenic acid A and kudinoside A as internal refererence substances (IRS), compatibility results showed that the relative correction factors (RCFs) of all compounds showed good reproducibility under different chromatographic conditions. On the basis of previous studies, the accuracy of the QAMS method was systematically evaluated by investigating the influences of curve intercept, analytes and IRS concentration. The results showed that the concentration (especially at low level) of analytes and curve intercept were the major influencing parameters for the LRG-QAMS method (LRG = linear regression), whereas the influence of IRS concentration seemed more apparent in terms of the AVG-QAMS method (AVG = average). The two approaches were complementary with each other. In addition, hierarchical clustering analysis (HCA), principal components analysis (PCA) and similarity analysis (SA) were performed to differentiate and classify the samples based on the contents of 18 marker compounds. The results of the different chemometric analyses were completely consistent with each other, and could be supported by the quantification results.

Ilex kudingcha C.J. Tseng tea and insect tea, as traditional Chinese teas, are favored for their original craftsmanship, unique flavor, and biological functionality. In this study, ultra high-performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-QqQ-MS) was used to analyze the bioactive components of the extracts of Ilex kudingcha and insect tea , and D-galactose-induced aging mice were used to compare the in vivo anti-aging effects of Ilex kudingcha and insect tea extracts. The results were remarkable, UHPLC-QqQ-MS analysis showed that ITP contains 29 ingredients, while IKDCP contains 26 ingredients. However, due to the large differences in the content of the main chemical components in IKDCP and ITP, the effects are equally different. At the same time, the in vivo research results suggesting that the anti-aging effects of IKDCP and ITP (500 mg/kg) include the regulation of viscera indices of major organs; improvement in liver, skin, and spleen tissue morphology; decreased production of inflammatory cytokines; up regulation of SOD, CAT, GSH, GSH-PX , and T-AOC and down regulation of NO and MDA levels in serum and liver tissue; reductions in the concentration of pro-inflammatory factors, and increases in the concentration of anti-inflammatory factor. RT-qPCR and western blot assay also showed that IKDCP and ITP affect anti-aging by regulating the gene and protein expression of GSH-PX, GSH1, SOD1, SOD2 , and CAT . The overall results indicate that ITP is more effective in treating oxidative damage in aging mice induced by D-galactose. Thus, ITP appears to be an effective functional drink owing to its rich nutritional components and anti-aging activities.

Autism spectrum disorders (ASD) have heterogeneous etiologies involving dysfunction of central nervous systems, for which no effective pan-specific treatments are available. Ilex kudingcha (IK) C.J. Tseng is a nootropic botanical used in Asia for neuroprotection and improvement of cognition. This study establishes that a chemically characterized extract from IK (IKE) mitigates behavioral traits in the Drosophila melanogaster rugose mutant, whose traits resemble human ASD, and examines possible mechanisms. IKE treatment significantly ameliorated deficits in social interaction, short-term memory, and locomotor activity in Drosophila rugose, and significantly increased synaptic bouton number of size more than 2 μm 2 in the neuromuscular junctions (NMJs) of Drosophila rugose . To clarify mechanism(s) of IKE action, methylphenidate (MPH), a dopamine transporter inhibitor, was included as a reference drug in the behavioral assays: MPH significantly improved social interaction and short-term memory deficit in Drosophila rugos e; administration of the dopamine D1 receptor antagonist SCH23390 and dopamine D2 receptor antagonist sulpiride reversed the ameliorative effects of both MPH and IKE on the social interaction deficits of Drosophila rugose . To extend analysis of IKE treatment to the vertebrate central nervous system, ASD-associated gene expression in mouse hippocampus was studied by RNA-seq: IKE treatment altered the expression of genes coding phosphoinositide 3-kinases/protein kinase B (PI3K-Akt), proteins in glutamatergic, dopaminergic, serotonergic, and GABAergic synapses, cAMP response element-binding protein (CREB), and RNA transporter proteins. These results provide a foundation for further analysis of IKE as a candidate for treatment of some forms of ASD.

Kuding tea, the leaves of Ilex Kudingcha C.J. Tseng, has been applied for treating obesity, hypertension, cardiovascular disease, hyperlipidemia, and so on. The chlorogenic acids (CGAs) in Kuding tea have shown excellent antioxidative, antiobesity, anti-atherosclerotic and anticancer activities. Nevertheless, the chemical profiles of CGAs in Kuding tea have not been comprehensively studied yet, which hinders further quality control. In the present study, a sensitive ultra-high-performance liquid chromatography-diode array detection coupled with a linear ion trap-Orbitrap (UHPLC-DAD-LTQ-Orbitrap) method was established to screen and identify CGAs in Kuding tea. Six CGA standards were first analyzed in negative ion mode with a CID-MS/MS experiment and then the diagnostic product ions (DPIs) were summarized. According to the retention behavior in the RP-ODS column, accurate mass measurement, DPIs and relevant bibliography data, a total of 68 CGA candidates attributed to 12 categories were unambiguously or preliminarily screened and characterized within 18 min of chromatographic time. This was the first systematic report on the distribution of CGAs in Kuding tea. Meanwhile, the contents of 6 major CGAs in Kuding tea were also determined by the UHPLC-DAD method. All the results indicated that the established analytical method could be employed as an effective technique for the comprehensive and systematic characterization of CGAs and quality control of the botanic extracts or Chinese medicinal formulas that contain various CGAs.

Trypanosoma cruzi causes chagas disease, a life threating disease in non-endemic and endemic regions globally, the life cycle of T. cruzi strictly depends on endogenous synthesis of sterol via 14-α-demethylase pathway. The available drugs for chagas disease treatment are currently resistance and parade unwanted side effects. Herein, molecular docking, QSAR, molecular mechanics/generalized born surface area (MM/GBSA) estimation, ADME screening, and molecular dynamics (MD) simulation were performed using Schrodinger suite to identify 14-α-demethylase protease antagonist from Ilex kudingcha . Density function theory of the hit ligands was carried out using Spartan 14 to investigate the molecular reactivity of the lead molecules. Nine (9) hit molecules were predicted as 14-α-demethylase protease inhibitors with binding energy range of − 7.632 to − 9.559 kcal/mol which was comparable to the standard drug (benznidazole = − 6.969), two lead molecules were further subjected to MD simulation over 50 ns predicting that kulactone and gallocatechin form stable interactions with vital residues at the catalytic site of the protein. DFT analysis revealed that, the hit ligands have the ability to donate and accept proton donating and accepting hence, effective as solubility and inhibitory agent and the ADME screening revealed, all the hit ligands obey Lipinski rule of five presenting them as drug candidate. The observations from this study predict kulactone and gallocatechin as putative antagonist of 14-α-demethylase protease and should be experimentally verify as a lead compound for chagas disease therapy.

The present study aimed to investigate the anti-inflammatory effects of Ilex kudingcha C.J. Tseng methanol extracts (KME) on 3% dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice (C57BL/6J strain). Body weight, disease activity index (DAI), colon length, colon weight to length ratio, colonic myeloperoxidase (MPO), glutathione (GSH) and malondialdehyde (MDA) levels were measured. Histological changes were observed by hematoxylin and eosin staining. Colonic levels of tumor necrosis factor-α (TNF-α), interleukin(IL)-1β and IL-6 were measured with an enzyme-linked immunosorbent assay. The mRNA expression of TNF-α, IL-1β, -6, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the colon tissue, was quantified by RT-PCR. KME significantly suppressed DSS-induced body weight loss, colon length shortening and decreased the colon weight to length ratio. It also resulted in increased GSH and reduced MPO and MDA levels in the colon tissue. Histological observation suggested that KME prevented edema, mucosal damage and loss of crypts, which are induced by DSS. In addition, KME decreased the levels of TNF-α, IL-1β and -6 in the colon tissues, while inhibiting the mRNA expression of these cytokines, as well as iNOS and COX-2. The results of this study suggested that KME has anti-inflammatory effects on DSS-induced UC in mice (C57BL/6J strain) by reducing the colonic levels and inhibiting the mRNA expression of pro-inflammatory cytokines.

Ilex kudingcha C.J. Tseng (kudingcha) is a traditional Chinese beverage widely consumed in East Asia. In the present study, the preventative effect of kudingcha on gastric injury was determined in Sprague-Dawley (SD) rats. High concentrations of kudingcha were observed to reduce the levels of the serum proinflammatory cytokines, interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α), compared with low concentrations of kudingcha. Gastric secretion volumes were highest in control rats and reduced in the following order; 250, 500 and 1,000 mg/kg kudingcha-treated and normal rats. The pH levels of gastric juice samples obtained from each group revealed the opposite correlation. Gastric injury levels in rats treated with kudingcha were identified to be significantly reduced, demonstrating its anti-inflammatory properties. Administration of 1,000 mg/kg kudingcha (73.9%) by gavage was demonstrated to induce the highest inhibitory effect on gastric injury. Results of the current study indicate that kudingcha exhibits marked preventive effects on gastric injury.

Tea and coffee are caffeinated beverages commonly consumed around the world in daily life. Tea from Camellia sinensis is widely available and is a good source of caffeine and other bioactive compounds (e.g., polyphenols and carotenoids). Other tea-like beverages, such as those from the genus Ilex, the large-leaved Kudingcha (Ilex latifolia Thunb and Ilex kudingcha C.J. Tseng), Yerba Mate (Ilex paraguariensis A. St.-Hil), Yaupon Holly (Ilex vomitoria), and Guayusa (Ilex guayusa Loes) are also traditional drinks, with lesser overall usage, but have attracted much recent attention and have been subjected to further study. This review summarizes the distribution, composition, and health benefits of caffeinated beverages from the genus Ilex. Plants of this genus mainly contain polyphenols and alkaloids, and show diverse health benefits, which, as well as supporting their further popularization as beverages, may also lead to potential applications in the pharmaceutical or nutraceutical industries.

Kuding tea (KT) is a bitter-tasting herbal tea that has been commonly used in traditional Chinese medicine (TCM). The large-leaved Ku-Ding-Cha (Aquifoliaceae) is composed of its representative species Ilex latifolia Thunb and Ilex kudingcha C.J. Tseng. Because of its potential lipid-lowering, body weight-reducing and blood-glucose-lowering properties, KT has increasingly been recognised for its importance over the past several decades. KT is no longer used only as a beverage, and various extraction methods have been applied to obtain aqueous and ethanolic KT extracts (KTE) or their fractions, which could potentially be used as dietary supplements. The major bioactive components of KT are triterpene saponins and polyphenols, but the composition of KT differs substantially between and among the different KT species. This in turn might affect the physiological effects of KT. KT exhibits antiobesity properties, possibly partly by affecting the intestinal microbiota. In addition, KT may mediate putative antioxidative, anti-inflammatory and anticancer activities. However, there is evidence that high KTE supplementation can adversely affect liver metabolism. The physiological relevance of KT in humans remains rather unclear since the potential health benefits of KT and its constituents reviewed here are mainly derived on the basis of in vitro and animal studies.