Ilex Kudingcha Research Articles

Health Benefits of Bioactive Compounds from the Genus Ilex, a Source of Traditional Caffeinated Beverages.

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.

Composition Analysis by UPLC-PDA-ESI (-)-HRMS and Antioxidant Activity Using Saccharomyces cerevisiae Model of Herbal Teas and Green Teas from Hainan.

Different teas from everywhere are very useful and have been extensively studied. We studied the antioxidant activity of herbal teas and green teas from Hainan, Mallotus oblongifolius Muell. Arg. (MO), Ilex kudingcha C.J. Tseng (KD), Camellia sinensis var. assamica (J. W. Mast.) Kitam. Hainan Dayezhong (DY), and Camellia sinensis (L.) O. Ktze. (produced from Hainan Baisha (BS)). The total phenol content and total flavonoid content from water extracts, resin extracts and fractions of herbal teas and green teas were compared. Later, eight fractions of herbal teas and green teas were subjected to UPLC-PDA-ESI-(−)-HRMS. We determined 1-diphenyl -2-picryl-hydrazyl radical and hydroxyl free radical scavenging activity by electron paramagnetic resonance spectroscopy. We subjected Saccharomyces cerevisiae to hydrogen peroxide, stress and evaluated antioxidant activity of herbal teas and green teas in cellulo. The experiment identified more than 14 potential antioxidant compounds from herbal teas and green teas. The herbal teas and green teas had a clearance rate higher than ferulic acid at the same concentrations. MO best reduced intracellular oxidation levels and increased catalase, glutathione reductase activities, glutathione reduced and glutathione oxidized content. KD had the highest cell survival rate and reduced cell lipid peroxidation. DY best improved superoxide dismutase activity and BS was the most active in the halo test. Therefore, we concluded that MO had stronger antioxidant activity than other herbal teas and green teas from Hainan, especially, which reduce S. cerevisiae oxidative stress under H2O2 stress.

Multi-Component Analysis of Ilex Kudingcha C. J. Tseng by a Single Marker Quantification Method and Chemometric Discrimination of HPLC Fingerprints.

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.

Simultaneous qualitative and quantitative evaluation of Ilex kudingcha C. J. tseng by using UPLC and UHPLC-qTOF-MS/MS

In this study, a systematic method was established for the holistic quality control of Ilex kudingcha C. J. Tseng, a popular functional drink for adjuvant treatment of diabetes, hypertension, obesity and hyperlipidemia. Both qualitative and quantitative analyses were conducted. For qualitative analysis, an ultra high performance liquid chromatography (UHPLC) coupled with an electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-qTOF-MS) method was established for rapid separation and structural identification of the constituents in Ilex kudingcha. Samples were separated on an ACQUITY UPLC HSS T3C18 column (2.1 mm × 100 mm, 1.8 μm) by gradient elution using 0.1% (v/v) formic acid (solvent A) and acetonitrile (solvent B) as mobile phases at a flow rate of 0.25 mL min−1. The chromatographic profiling of Ilex kudingcha by UHPLC-qTOF-MS/MS resulted in the characterization of 53 compounds, comprising 18 compounds that were unambiguously identified by comparison with reference standards. For quantitative analysis, 18 major compounds from 15 batches of Ilex kudingcha samples were simultaneously detected by UPLC-DAD at wavelengths of 210 nm, 260 nm, and 326 nm. The method was validated with respect to precision, linearity, repeatability, stability, accuracy, and so on. The contents of the 18 target compounds were applied for hierarchical clustering analysis (HCA) and principal component analysis (PCA) to differentiate between the samples. The results of HCA and PCA were consistent with each other. Sample No. 1 differed significantly based on HCA and PCA, and the differentiating components were confirmed to originate from different batches of samples. Phenolic acids and triterpenes were found to be the main ingredients in Ilex kudingcha. This strategy was effective and straightforward, and provided a potential approach for holistic quality control of Ilex kudingcha.

Kudingcha and Fuzhuan Brick Tea Prevent Obesity and Modulate Gut Microbiota in High‐Fat Diet Fed Mice

Kudingcha (KDC) from Ilex kudingcha and Fuzhuan brick tea (FBT) are popular beverages in China, and their preventive and therapeutic roles in metabolic disorders have been reported. However, the relationship between the gut microbiota modulatory effects of KDC and FBT and prevention of obesity is still not clearly understood. KDC and FBT are tested individually for their capacities to prevent obesity and modulate the gut microbiota in high-fat diet (HFD) fed C57BL/6J mice. The results show that both KDC and FBT supplementation could modulate oxidative injury, inflammation, lipid metabolism, and reduce HFD induced obesity significantly. Both KDC and FBT could enhance the diversity of gut microbiota. KDC could reduce the relative abundance of Erysipelotrichaceae, while FBT could reduce the ratio of Firmicutes to Bacteroidetes and enhance the relative abundance of Bifidobacteriaceae. These findings suggest that KDC and FBT could attenuate features of the metabolic syndrome in HFD-fed mice, which might be due to the modulation of gut microbiota by KDC and FBT.

Histopathological Studies on the Effects of Chloroform and Methanolic Extracts of Ilex kudingcha in Trypanosoma brucei Infected Albino Wistar Rats

Histopathological studies of the effects of chloroform and methanolic leaf extracts of Ilex kudingcha in Trypanosoma brucei infected albino wistar rats were investigated. The toxicity and phytochemical study were also carried out using standard protocol. T. brucei infected animals were administered orally with 200 and 400 mg/kg b.w. of the extracts and 3.5 mg/kg b.w. of the standard drug (diminazene aceturate). Results on acute toxicity studies (LD50) revealed no sign of lethality up to the dose of 5,000 mg/kg body weight but the liver and kidney histology of infected animals treated with 5,000 mg/kg b.w. of I. kudingcha extracts were observed to be hepatotoxic and nephrotoxic. The methanol extracts showed appreciably high in vivo anti-trypanosomal activities compared to the reference drug. Histological examination of the organs revealed serious pathological lesions in the liver of the infected animals without treatment (negative control). In the positive control animals (infected animals administered standard drug), mild multifocal aggregate of inflammatory leucocytes was observed. In the other experimental animals, no pathological lesion was observed in the liver, kidney, brain, and heart of infected animals treated with the methanolic extract and combined methanol and chloroform extracts. The effectiveness of the methanolic extract at reducing the lesions caused by the parasite is the same compared with the standard drug. Phytochemical analysis of the plant extracts showed that methanol extract contained appreciable high levels of alkaloids, saponin, tannins, phenol, and glycoside while flavonoid was not detected. Hence, the curative properties of methanolic extract of I. kudingcha as observed in the organs indicate its anti-trypanosomal properties but it should be consumed at minimal doses.

Chlorogenic Acid-Enriched Extract of Ilex kudingcha C.J. Tseng Inhibits Angiogenesis in Zebrafish.

Kudingcha is a particularly bitter tasting tea that has been widely used in China to eliminate fever and itching eyes, and to clear blood toxins. Kudingcha is considered of value for its potential anticancer effects that are attributed to the presence of characteristic bioactive ingredients. The chlorogenic acid (CGA) derivatives 3-0-caffeoylquinic acid, 5-0-caffeoylquinic acid, 3,5-0-dicaffeoylquinic acid, and 4,5-0-dicaffeoylquinic acid were separated from Ilex kudingcha C.J. Tseng extract by high-performance liquid chromatography (HPLC)-photodiode array detector (PDA) and HPLC-nuclear magnetic resonance (NMR). In Tg(flk1:EGFP) zebrafish embryos at 52 hours postfertilization (hpf), angiogenesis was significantly inhibited by kudingcha extract (KDCE) at concentrations of 400 and 500 μg/mL and CGA also showed significant inhibition in embryos treated with 80, 100, and 130 μg/mL. Endothelial cell apoptosis showed a dose-dependent increase in response to KDCE and CGA. CGA derivatives from KDCE could have potential as anticancer agents against tumor angiogenesis.

Kudinoside-D, a triterpenoid saponin derived from Ilex kudingcha suppresses adipogenesis through modulation of the AMPK pathway in 3T3-L1 adipocytes

The leaves of Ilex Kudingcha, locally named “Kudingcha” in China, has been traditionally applied for treating obesity. Studies have demonstrated that the ethanol extract of Ilex kudingcha have anti-adipogenic effects. However, the constituent which was responsible for its anti-obesity and its underlying molecular mechanism has not yet been elucidated. This research explored the anti-obesity effect of kudinoside-D which was a main natural component of triterpenoid saponin from the ethanol extract of Ilex kudingcha, on lipid accumulation and the potential mechanism of action of adipogenesis in 3T3-L1 adipocytes. The adipocytes were treated with various concentrations of kudinoside D (0 to 40μM) during differentiation. The image-based Oil Red O staining analyses revealed that KD-D, dose dependently reduced cytoplasmic lipid droplet in 3T3-L1 adipocytes with the IC50 is 59.49μM. Meanwhile, major adipogenic transcription factor peroxisome proliferator-activated receptor γ (PPARγ), CCAAT/enhancer binding protein-α (C/EBPα) and sterol regulatory element-binding protein 1c (SREBP-1c) were significantly repressed as well as their target genes. The phosphorylation of AMP-activated protein kinase (AMPK) and its downstream target phosphorylated-acetyl CoA carboxylase (ACC) expression were also increased. In addition, the inhibitory effects of KD-D on the expressions of PPARγ and C/EBPα were weakened when cells were cotreated with AMPK inhibitor Compound C. These results indicated KD-D exerts anti-adipogenic effects through modulation of adipogenic transcription factors via AMPK signaling pathway. And the current findings demonstrated that KD-D was a potential therapeutic candidate for alleviating obesity and hyperlipidemia.

Modulating Effects of Dicaffeoylquinic Acids from Ilex kudingcha on Intestinal Microecology in Vitro.

Dietary polyphenols have been considered as novel prebiotics, and polyphenols could exert their functions through modulating intestinal microbiota. The diverse bioactivities of kudingcha could derive from its phenolic compounds, but the effects of dicaffeoylquinic acids (diCQAs) from Ilex kudingcha on intestinal microbiota have not been investigated. In the present study, high-throughput sequencing and anaerobic fermentation in vitro were utilized to investigate the microecology-modulating function of I. kudingcha diCQAs. As a result, diCQAs raised the diversity and exhibited a more considerable impact than a carbon source on the microbial profile. DiCQAs increased the relative abundances of Alistipes, Bacteroides, Bifidobacterium, Butyricimonas, Clostridium sensu stricto, Escherichia/Shigella, Parasutterella, Romboutsia, Oscillibacter, Veillonella, Phascolarctobacterium, Lachnospiracea incertae sedis, Gemmiger, Streptococcus, and Haemophilus and decreased the relative abundances of Ruminococcus, Anaerostipes, Dialister, Megasphaera, Megamonas, and Prevotella. DiCQAs also affected the generation of short-chain fatty acids through microbiota. The contents of acetic and lactic acids were raised, while the production of propionic and butyric acids was reduced. Conclusively, diCQAs from I. kudingcha had significant modulating effects on intestinal microbiota in vitro, which might be the fundamental of diCQAs exerting their bioactivities.

Inhibition effect of Ilex kudingcha C.J. Tseng (Kudingcha) extract on J55 Steel in 3.5wt% NaCl Solution Saturated with CO2

Inhibition effect of Ilex kudingcha C.J. Tseng (Kudingcha) extract on J55 Steel in 3.5wt% NaCl Solution Saturated with CO2

Simultaneous Qualitation and Quantitation of Chlorogenic Acids in Kuding Tea Using Ultra-High-Performance Liquid Chromatography-Diode Array Detection Coupled with Linear Ion Trap-Orbitrap Mass Spectrometer.

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.

Hydrolysis of Dicaffeoylquinic Acids from Ilex kudingcha Happens in the Colon by Intestinal Microbiota.

Monocaffeoylquinic acids (mono-CQAs) can be hydrolyzed or metabolized by pancreatin, intestinal brush border esterase, and microbiota in the colon. Data about the conversion of dicaffeoylquinic acids (diCQAs) in digestion are scarce. The diCQA-rich fraction including 3,4-, 3,5-, and 4,5-diCQAs was prepared from Ilex kudingcha, and the conversion in simulated gastricintestine was investigated. Artificial saliva, gastric and pancreatic fluids, Caco-2 monolayer cells, and anaerobic fermentation model were utilized to mimic digestions of the oral cavity, stomach, small intestine, and colon in vitro. The results revealed that diCQAs remained intact in simulated saliva, gastric, and pancreatic fluids and within Caco-2 cells. In anaerobic fermentation with human fecal slurry, diCQAs were hydrolyzed to mono-CQAs and caffeic acid, which were further metabolized to caffeic acid and dihydrocaffeic acid, respectively. The hydrolysis of diCQAs depended on the chemical structures, carbohydrates in the culture medium, and microbial compositions. Our research demonstrated that hydrolysis of diCQAs happened in the colon by intestinal microbiota.

Triterpenoid-enriched extract of Ilex kudingcha inhibits aggregated LDL-induced lipid deposition in macrophages by downregulating low density lipoprotein receptor-related protein 1 (LRP1)

Macrophage-derived foam cell formation is considered to be a critical step in the development of atherosclerosis. In the present study, we evaluated the inhibitory effect of Ilex kudingcha triterpenoid-enriched extract (TS) on foam cell formation in THP-1 macrophage cells induced by aggregated low density lipoproteins (agLDL). The inhibition of low density lipoprotein receptor-related protein 1 (LRP1) expression after TS treatment is based on mRNA and protein levels in the macrophages. These inductions were sterol regulatory element-binding protein-2 (SREBP2) dependent. On the basis of the in vitro results, the anti-atherosclerotic activity of TS was investigated in ApoE−/− mice fed a high-fat diet, ApoE−/− mice treated with TS for 6 weeks appeared to have decreased aortic sinus lesions compared with the untreated mice. These findings indicate that TS may act as an anti-atherosclerotic agent.

Antiplatelet aggregation triterpene saponins from the leaves of Ilex kudingcha

Two new ursane-type triterpene saponins, 3-O-β-d-glucopyranosyl(1→3)-[α-l-rhamnopyranosyl(1→2)]-α-l-arabinopyranosylurs-12,19(29)-dien-28-oic acid 28-O-α-l-rhamnopyranosyl(1→2)-β-d-glucopyranosyl ester (1) and 3-O-β-d-glucopyranosyl(1→3)-[α-l-rhamnopyranosyl(1→2)]-α-l-arabinopyranosyl-19α,20α-dihydroxyurs-12-en-28-oic acid 28-O-α-l-rhamnopyranosyl(1→2)-β-d-glucopyranosyl ester (2), along with thirteen known triterpene saponins were isolated from the n-BuOH part of the MeOH extraction of the leaves of Ilex kudingcha C.J. Tseng (also called “Ku-Ding-Cha”). The structures of new compounds were elucidated on the basis of detailed spectroscopic analysis, including HR-ESI-TOF-MS, 1D and 2D-NMR experiments, and by acid hydrolysis. All the compounds were screened for antiplatelet aggregation activity in vitro, and compounds 1, 2, 3, 7, 12 and 15 showed significant inhibition of platelet aggregation induced by ADP (5μM) with IC50 values of 14.7±3.7, 11.3±2.5, 17.4±4.6, 20.5±3.1, 8.1±1.5 and 18.9±4.2μM, respectively.

Inhibitory Activities of Caffeoylquinic Acid Derivatives from Ilex kudingcha C.J. Tseng on α-Glucosidase from Saccharomyces cerevisiae

Polyphenols and caffeoylquinic acid (CQA) derivatives (3-CQA, 4-CQA, 5-CQA, 3,4-diCQA, 3,5-diCQA, and 4,5-diCQA) were prepared from Ilex kudingcha C.J. Tseng, and their effects and mechanisms on the activities of α-glucosidase from Saccharomyces cerevisiae were investigated in the present study. As results, the IC50 values for CQA derivatives were 0.16-0.39 mg/mL, and the inhibition mode of CQA derivatives was noncompetitive. On the basis of fluorescence spectroscopy and circular dichroism spectroscopy data, the binding constants and number of binding sites were calculated to be 10(6)-10(8) M(-1) and 1.42-1.87, respectively. CQA derivatives could bind to the enzyme mainly through hydrophobic interaction, altering the microenvironment and molecular conformation of the enzyme, thus decreasing the catalytic activity. To the authors' knowledge, this is the first report on α-glucosidase inhibitory mechanism by CQA derivatives from I. kudingcha, and the findings suggest a potential use of kudingcha as functional foods for the prevention and treatment of diabetes and related symptoms.

In vitro and in vivo anti-cancer activities of Kuding tea (Ilex kudingcha C.J. Tseng) against oral cancer

Kuding tea (Ilex kudingcha C.J. Tseng) is drunk widely in China. The in vitro anticancer effects of Kuding tea were evaluated in TCA8113 human tongue carcinoma cells using a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay. At a concentration of 200 μg/ml, Kuding tea exhibited an inhibitory effect of 75% in TCA8113 cells, which was higher than that observed at concentrations of 100 and 50 μg/ml (41 and 10% inhibition, respectively). Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analyses of the apoptosis, inflammation and metastasis genes and proteins in Kuding tea-treated cancer cells were performed. Kuding tea significantly induced apoptosis in TCA8113 cancer cells (P<0.05) by upregulating Bax, caspase-3 and caspase-9 expression, and downregulating Bcl-2 expression. Expression of the NF-κB, iNOS and COX-2 genes that are associated with inflammation was significantly downregulated by Kuding tea, which demonstrated its anti-inflammatory properties. Kuding tea also exerted an anti-metastatic effect on cancer cells. This was demonstrated by the decreased expression of matrix metalloproteases (MMPs) and the increased expression of tissue inhibitors of metalloproteinases (TIMPs), and confirmed by the inhibition of the metastasis of U14 squamous cell carcinoma cells in imprinting control region (ICR) mice. The ICR mouse buccal mucosa cancer model was established by injecting the mice with U14 cells. Following injection, the wound at the injection site was topically treated with Kuding tea. It was observed that the tumor volumes for the group treated with Kuding tea were smaller than those from the control mice. Analysis of the sections of buccal mucosa cancer tissue demonstrated that the buccal mucosa cancer degrees of the Kuding tea-treated mice were weaker than that in the control mice. Similar results were observed in the lesion sections of the cervical lymph nodes. Based on these results, Kuding tea exhibited successful in vitro anticancer effects in TCA8113 cells and in vivo buccal mucosa cancer preventive activity.

Fortification of the Health Benefit of Buckwheat (F agopyrum tataricum ) Tea

Three extracts from Zijuan (Camellia sinensis), Kudingcha (Ilex kudingcha) and Mingri (Angelica keiskei) leaves were used to fortify the health promoting function of traditional buckwheat tea. Three fortified teas with similar sensory property to the original tea and higher level of total flavonoids and catechin were chosen to evaluate their capability in lowering hyperglycemic and hyperlipidemic stress using mouse models. The blood glucose level in the hyperglycemic mice was significantly reduced by 10.84 to 34.12% after oral gavage of the three fortified teas (0.4 mL of 20% in solution) each day for 7 days, while it only lowered 4.56% in the original buckwheat tea treatment (P < 0.05). Also, the three fortified teas had effective response to lower the boosted blood glucose level in fasted mice. In the hyperlipidemic mouse model, the mice fed with the fortified teas for 30 days had lower serum total cholesterol and triglyceride levels and higher high-density lipoprotein-cholesterol level. Practical Applications This study demonstrated a potential method to enhance the health benefit of buckwheat tea without loss of its sensory properties. The information would be valuable in developing various health-promoting food products from buckwheat and other grains.

Study on the Phytoavailability of Heavy Metal Lead, Copper, Zinc in Ilex Kudingcha C. J. Tseng Soil

Pot experiment was conducted with the cutting seedlings of Ilex kudingcha C. J. Tseng. By adding lead, copper solution and self-exist zinc in soil, we conducted the research for the phytoavailability of heavy metal in the different soil situation. The characterization methods of the phytoavailability were investigated. Results show that lead, copper and zinc are the strong absorption elements from BAC (Biological Absorption Coefficient) averages of 13 groups experiment. The order of BAC averages is lead &gt; copper &gt; zinc. 0.05mol/L EDTA-2Na can be used as the extracting agent of lead or copper, 0.1mol/L HCl can be used as the extracting agent of zinc. They can be used for the characterization methods about the phytoavailability of lead, copper, zinc in soil by the single extraction method. 0.1mol/L HC1 is adopted to extract soil, and weighted ion impulse of the lead, copper, zinc in the extraction fluid is calculated, which can well simultaneously characterize the phytoavailability of heavy metal lead, copper, zinc in kudingcha soil.

Anti-inflammatory effects of kudingcha methanol extract (Ilex kudingcha C.J. Tseng) in dextran sulfate sodium-induced ulcerative colitis

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) prevents HCl/ethanol-induced gastric injury in Sprague-Dawley rats

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 1,000mg/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 1,000mg/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.