Phillyrin ameliorates influenza a virus-induced pulmonary inflammation by antagonizing CXCR2 and inhibiting NLRP3 inflammasome activation.

Influenza is an acute viral respiratory illness with high morbidity rates worldwide. Excessive pulmonary inflammation is the main characteristic of lethal influenza A virus (IAV) infections. Therapeutic options for managing influenza are limited to vaccines and some antiviral medications. Phillyrin is one of the major bioactive components of the Chinese herbal medicine Forsythia suspensa, which has the functions of sterilization, heat clearing and detoxification. In this work, the effect and mechanism of phillyrin on H1N1 influenza (PR8)-induced pneumonia were investigated. We reported that phillyrin (15 mg/kg) treatment after viral challenge significantly improved the weight loss, ameliorated pulmonary inflammation and inhibited the accumulation of multiple cytokines and chemokines in bronchoalveolar lavage fluid on 7 days post infection (dpi). In vitro, phillyrin suppressed influenza viral replication (Matrixprotein and nucleoprotein messenger RNA level) and reduced influenza virus-induced cytopathic effect (CPE). Furthermore,chemokine receptor CXCR2 was confirmed to be markedly inhibited by phillyrin. Surface plasmon resonance results reveal that phillyrin exhibits binding affinity to CXCR2, having a binding affinity constant (KD) value of 1.858e-5 M, suggesting that CXCR2 is a potential therapeutic target for phillyrin. Moreover, phillyrin inhibited the mRNA and protein expression levels of Caspase1, ASC and NLRP3 in the lungs of mice with H1N1-induced pneumonia.This study reveals that phillyrin ameliorates IAV-induced pulmonary inflammation by antagonizing CXCR2 and inhibiting NLRP3 inflammasome activation partly.

Decompression Mechanism of Radish Seed in Prehypertension Rats through Integration of Transcriptomics and Metabolomics Methods.

Radish seed (RS), the dried ripe seed of Raphanus sativus L., is widely used in traditional Chinese medicine (TCM) to reduce blood pressure. However, the molecular and pharmacological mechanisms underlying its therapeutic effects are still unclear. In this study, we analyzed the effects of RS in a rat model of prehypertension and assessed the mechanistic basis by integrating transcriptomics and metabolomics. RS administration significantly reduced blood pressure in prehypertensive male Wistar rats, negatively regulated endothelin-1, increased nitric oxide levels, and reduced the exfoliation of endothelium cells. In vitro vascular ring experiments further confirmed the effects of RS on vascular endothelial cells. Furthermore, we identified 65 differentially expressed genes (DEGs; P adj < 0.05 and fold change (FC) > 2) and 52 metabolites (VIP > 1, P < 0.05 and FC ≥ 2 or ≤0.5) in the RS intervention group using RNA-seq and UPLC-MS/MS, respectively. A network of the DEGs and the metabolites was constructed,q which indicated that RS regulates purine metabolism, linoleic acid metabolism, arachidonic acid metabolism, circadian rhythm, and phosphatidylinositol signaling pathway, and its target genes are Pik3c2a, Hspa8, Dnaja1, Arntl, Ugt1a1, Dbp, Rasd1, and Aldh1a3. Thus, the antihypertensive effects of RS can be attributed to its ability to improve vascular endothelial dysfunction by targeting multiple genes and pathways. Our findings provide new insights into the pathological mechanisms underlying prehypertension, along with novel targets for the prevention and treatment of hypertension.

The Mechanisms Underlying the Pharmacological Effects of GuiPi Decoction on Major Depressive Disorder based on Network Pharmacology and Molecular Docking

BACKGROUND AND AIM: Major Depressive Disorder (MDD) is a common affective disorder. GuiPi decoction (GPD) is used to treat depression in China, Japan, and Korea. However, its effective ingredients and antidepressant mechanisms remain unclear. We attempted to reveal the potential mechanisms of GPD in the treatment of MDD by network pharmacology and molecular docking. In addition, we conducted an enzymatic activity assay to validate the results of molecular docking. METHODS: GPD-related compounds and targets, and MDD-related targets were retrieved from databases and literature. The herb-compound-target network was constructed by Cytoscape. The protein- protein interaction network was built using the STRING database to find key targets of GPD on MDD. Enrichment analysis of shared targets was analyzed by MetaCore database to obtain the potential pathway and biological process of GPD on MDD. The main active compounds treating MDD were screened by molecular docking. The PDE4s inhibitors were screened and verified by an enzyme activity assay. RESULTS: GPD contained 1222 ingredients and 190 potential targets for anti-MDD. Possible biological processes regulated by GPD were neurophysiological processes, blood vessel morphogenesis, Camp Responsive Element Modulator (CREM) pathway, and Androgen Receptor (AR) signaling crosstalk in MDD. Potential pathways in MDD associated with GPD include neurotransmission, cell differentiation, androgen signaling, and estrogen signaling. Fumarine, m-cresol, quercetin, betasitosterol, fumarine, taraxasterol, and lupeol in GPD may be the targets of SLC6A4, monoamine oxidase A (MAOA), DRD2, OPRM1, HTR3A, Albumin (ALB), and NTRK1, respectively. The IC50 values of trifolin targeting Phosphodiesterase (PDE) 4A and girinimbine targeting PDE4B1 were 73.79 µM and 31.86 µM, respectively. The IC50 values of girinimbine and benzo[a]carbazole on PDE4B2 were 51.62 µM and 94.61 µM, respectively. CONCLUSION: Different compounds in GPD may target the same protein, and the same component in GPD can target multiple targets. These results suggest that the effects of GPD on MDD are holistic and systematic, unlike the pattern of one drug-one target.

Effects of Qi-Fu-Yin on aging of APP/PS1 transgenic mice by regulating the intestinal microbiome.

Introduction: Alzheimer's disease is the most common form of dementia and closely related to aging. Qi-Fu-Yin is widely used to treat dementia, but its anti-aging effects is unknown. Methods: We used 11-month-old APP/PS1 transgenic mice for behavioral tests to observe the changes in cognitive function and age-related symptoms after Qi-Fu-Yin treatment. Fecal samples were collected for 16sRNA sequencing and metagenomic sequencing. Differences among the groups of intestinal microbiota and the associations with aging and intestinal microbiota were analyzed based on the results. Results: Here we found that Qi-Fu-Yin improved the ability of motor coordination, raised survival rate and prolonged the survival days under cold stress stimulation in aged APP/ PS1 transgenic mice. Our data from 16sRNA and metagenomic sequencing showed that at the Family level, the intestinal microbiota was significantly different among wild-type mice, APP/PS1 transgenic mice and the Qi-Fu-Yin group by PCA analysis. Importantly, Qi-Fu-Yin improved the functional diversity of the major KEGG pathways, carbohydrate-active enzymes, and major virulence factors in the intestinal flora of APP/PS1 transgenic mice. Among them, the functions of eight carbohydrate-active enzymes (GT2_Glycos_transf_2, GT4, GT41, GH2, CE1, CE10, CE3, and GH24) and the functions of top three virulence factors (defensive virulence factors, offensive virulence factors and nonspecific virulence factors) were significantly and positively correlated with the level of grasping ability. We further indicated that the Qi-Fu-Yin significantly reduced the plasma levels of IL-6. Conclusion: Our results indicated that the effects of Qi-Fu-Yin anti-aging of APP/PS1 transgenic mice might be through the regulation of intestinal flora diversity, species richness and the function of major active enzymes.

Immune Effect of Active Components of Traditional Chinese Medicine on Triple-Negative Breast Cancer.

Triple-negative breast cancers are heterogeneous, poorly prognostic, and metastatic malignancies that result in a high risk of death for patients. Targeted therapy for triple-negative breast cancer has been extremely challenging due to the lack of expression of estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Clinical treatment regimens for triple-negative breast cancer are often based on paclitaxel and platinum drugs, but drug resistance and side effects from the drugs frequently lead to treatment failure, thus requiring the development of new therapeutic platforms. In recent years, research on traditional Chinese medicine in modulating the immune function of the body has shown that it has the potential to be an effective treatment option against triple-negative breast cancer. Active components of herbal medicines such as alkaloids, flavonoids, polyphenols, saponins, and polysaccharides have been shown to inhibit cancer cell proliferation and metastasis by activating inflammatory immune responses and can modulate tumor-related signaling pathways to further inhibit the invasion of triple-negative breast cancer. This paper reviews the immunomodulatory mechanisms of different herbal active ingredients against triple-negative breast cancer and provides an outlook on the challenges and directions of development for the treatment of triple-negative breast cancer with herbal active ingredients.

Toward the Development of Personalized Syndrome Discriminant Systems: A Discriminant System for Hypertension with Liver Yang Hyperactivity Syndrome.

Traditional Chinese medicine has shown promising results in treating the symptoms of hypertension, a major global health concern not yet fully managed by modern medicine. It is, therefore, of high priority to clarify the altered pathophysiology of hypertension in individuals with liver Yang hyperactivity syndrome (HLYH) in response to effective treatments to better understand this disorder. The primary aim of this study was to construct a personalized syndrome discriminant system based on data capable of informing management strategies prior to the initiation of antihypertensive therapy or the implementation of screening strategies in at-risk HLYH. Based on the successful replication of HLYH rat models, we extracted the core discriminant factors of the disorder through the integration of physical signs, biochemical indicators, and metabolic markers. Macro and micro information was correlated to construct a syndrome discriminant system. At the macroscopic level, HLYH rat models characterized by elevated blood pressure were found to be associated with significant changes in water intake, pain threshold, retention time on a rotating platform, and body surface temperature. A total of 27 potential biomarkers and 14 metabolic pathways appeared to reflect the primary metabolic characteristics. Through the integration of these data, we successfully constructed a combined macro-micro personalized syndrome discriminant system, which provides a foundation for research regarding the risk loci of HLYH. Our findings also broaden our understanding of the biological pathways involved in HLYH.

The Saggy Ink Cap Medicinal Mushroom, Coprinus comatus (Agaricomycetes), Protein Attenuates Acute Alcoholic Liver Injury in Association with Changes in the Gut Microbiota of Mice.

This study aims to investigate the hepatoprotective effects of Coprinus comatus protein (CCP) in a mouse model of acute alcoholic liver injury by regulating gut microbiota dysbiosis. Mice were divided into four groups, including the control group (CG), alcohol group (AG), biphenyldicarboxylate group (BG), and protein group (PG). The results showed that alcohol can increase the liver organ index, which could be adjusted by CCP. At the same time, analysis of serum biochemical indexes (alanine aminotransferase, aspartate aminotransferase) and liver oxidative stress levels (glutathione) revealed that CCP significantly alleviated alcohol-induced hepatic inflammation. Sequencing of 16S rRNA showed that gut microbiota composition was changed significantly by alcohol treatment. However, CCP could mitigate dysbiosis of gut microbiota, such as increasing the proportion of Muribaculaceae, Lachnospiraceae, and Lactobacillaceae and reducing the proportion of Burkholderiaceae, Deferribacteraceae, Enterococcaceae, and Enterobacteriaceae. In conclusion, CCP can maintain gut microbiota stability to improve liver injury and is potentially a good candidate for dietary supplements against acute alcoholic liver injury.

Clickable amino acid derivative tuned self-assembly of antigen and adjuvant for cancer immunotherapy.

Amino acid-tuned self-assembly has become an attractive strategy for constructing various functional materials. Here, a series of dibenzocyclooctyne (DIBO) functionalized amphiphilic amino acid derivatives are designed and screened as building blocks of functional supramolecular self-assembly nanoparticles for cancer immunotherapy. One top-performing supramolecular self-assembly material (named DA6C1) is identified through combinatorial screening, and spherical nanoparticles can be easily prepared by this material tuned multicomponent synergistic self-assembly of ovalbumin (OVA) and CpG oligonucleotide. DA6C1 based nanovaccine can significantly enhance the cellular uptake of OVA and CpG into the same bone marrow derived dendritic cells (BMDCs) and greatly improve the activation of DCs. Moreover, after subcutaneous injection, this nanovaccine flows rapidly to the lymph nodes and elicits strong immune responses to achieve effective prophylactic and therapeutic effect. Therefore, our work highlights the great potential of clickable amino acid derivatives as a convenient and powerful tool to construct nanovaccine for effective immunotherapy.

A network pharmacology-based study on key pharmacological pathways and targets of Qi Fu Yin acting on Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is a central nervous system (CNS) disease characterized by progressive cognitive dysfunction and memory loss. Qi Fu Yin is mainly used to treat dementia, particularly AD, in the clinic, but its comprehensive mechanisms are not known. OBJECTIVE: In this research, we aimed to investigate the mechanisms of Qi Fu Yin in AD by network pharmacology and molecular docking. METHODS: First, the chemical constituents in Qi Fu Yin were obtained from five databases and classified according to their structure. Targets of chemical constituents and AD-related targets were also collected from the databases. Then, overlapping genes between Qi Fu Yin and AD were identified by intersection analysis. MetaCore was used to gather enrichment information. Combination synergy analysis was performed by Cytoscape. After ligand-receptor docking, the binding affinity was verified by ADP-Glo™ kinase assay and fluorescence resonance energy transfer (FRET) assay. RESULTS: We found 12 classes with 977 components in Qi Fu Yin. A total of 511 compounds and 577 potential target proteins in Qi Fu Yin were found to be related to AD. The pathways of Qi Fu Yin in AD included oxidative stress and immune response. There was the best binding affinity between 11 pairs of genes and compounds. Furthermore, CDK5 was inhibited by nepetin with an IC50 of 3.172 µM and kaempferol with an IC50 of 2.659 µM. Ceanothic acid and 18 beta-glycyrrhetinic acid inhibited GSK3ß, and the IC50 values were 8.732 µM and 8.06 µM, respectively. CONCLUSION: Qi Fu Yin might alleviate Tau hyperphosphorylation by nepetin, kaempferol, ceanothic acid and 18 beta-glycyrrhetinic acid.

Identification of Chemical Components of Qi-Fu-Yin and Its Prototype Components and Metabolites in Rat Plasma and Cerebrospinal Fluid via UPLC-Q-TOF-MS.

Qi-Fu-Yin, a traditional Chinese medicine formula, has been used to treat Alzheimer's disease (AD, a neurodegenerative disorder) in clinical setting. In this study, the chemical components of Qi-Fu-Yin and its prototype components and metabolites in rat plasma and cerebrospinal fluid, after oral administration, were preliminarily characterized via ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS). A total of 180 compounds, including saponins, flavonoids, organic acids, sucrose esters, oligosaccharide esters, phthalides, phenylethanoid glycosides, alkaloids, xanthones, terpene lactones, ionones, and iridoid glycoside, were tentatively characterized. For the first time, 51 prototypical components and 26 metabolites, including saponins, phthalides, flavonoids, sucrose esters, organic acids, alkaloids, ionones, terpene lactones, iridoid glycoside, and their derivatives, have been tentatively identified in the plasma. Furthermore, 10 prototypical components (including butylidenephthalide, butylphthalide, 20(S)-ginsenoside Rh1, 20(R)-ginsenoside Rh1, and zingibroside R 1) and 6 metabolites were preliminarily characterized in cerebrospinal fluid. These results were beneficial to the discovery of the active components of Qi-Fu-Yin anti-AD.

Ameliorative effects of ginsenosides on myelosuppression induced by chemotherapy or radiotherapy.

BACKGROUND: and ethnopharmacological relevance: As the major side effect of radiotherapy or chemotherapy, myelosuppression usually leads to anemia, hemorrhage, immunosuppression, and even fatal infections, which may discontinue the process of cancer treatment. As a result, more and more attention is paid to the treatment of myelosuppression. Ginseng, root of Panax ginseng Meyer (Panax ginseng C. A. Mey), is considered as the king of herbs in the Orient, particularly in China, Korea and Japan. Ginsenosides, the most important active ingredients of ginseng, have been shown to have a variety of therapeutic effects, such as neuroprotective, anti-cancer and anti-diabetic properties. Considering that ginsenosides are closely associated with the pathogenesis of myelosuppression, researchers have carried out a few experiments on ginsenosides to attenuate myelosuppression induced by chemotherapy or radiotherapy in recent years. AIM OF THE STUDY: To summarize previous studies about the effects of ginsenosides on alleviating myelosuppression and the mechanisms of action. METHODS: Literatures in this review were searched in PubMed, China National Knowledge Infrastructure (CNKI), Web of Science, and ScienceDirect. RESULTS: Ginsenosides play an important role in relieving myelosuppression predominantly by restoring hematopoiesis and immunity. CONCLUSION: Ginsenosides might be potential candidates for the treatment of myelosuppression induced by chemotherapy or radiotherapy.

The Prebiotic-Like Effects of Coprinus comatus Polysaccharides on Gut Microbiota in Normal Mice and Those with Acute Alcoholic Liver Injury: A Comparative Study.

This study aims to investigate the prebiotic-like effects of Coprinus comatus polysaccharides (CCP) on gut microbiota. Mice were divided into four groups: normal group (NG), alcohol group (AG), polysaccharides group (PG), and alcohol + polysaccharides group (APG). The gut microbiota structure of feces was analyzed by determining the V3-V4 region sequence in 16S rDNA. The results showed CCP could increase the diversity of gut microbiota. Compared with NG, PG had a significantly higher relative abundance of Firmicutes and Lactobacillaceae and a lower abundance of Rikenellaceae. These changes in gut microbiota result in positive effects on gut due to a series of prebiotic-like effects of CCP. At the same time, CCP could improve some adverse changes in gut microbiota caused by acute alcohol intake, such as the increased proportion of Firmicutes, Bacteroidetes, Muribaculaceae, and Lachnospiraceae and the decreased proportion of Rikenellaceae. In conclusion, the CCP has certain prebiotic effects not only on normal mice but also on mice with acute alcoholic liver injury.

A novel time-dimension and circadian rhythm-dependent strategy for pharmacodynamic evaluation of Uncaria in the regulation of neurotransmitter circadian metabolic homeostasis in spontaneously hypertensive rats.

In the present study, we aimed to use metabolomics platforms to examine circadian-regulated neurotransmitters across a 24-h day and the effects of Uncaria administration on daily rhythmicity in order to establish a strategy for evaluating the spatiotemporal efficacy evaluation strategy of Uncaria. By using targeted ultrahigh performance liquid chromatography-mass spectrometry metabolomics, we quantified 32 neurotransmitter metabolites every 4 h over 24 h. To assess 24-h metabolite rhythmicity, we performed cosinor analysis. The expression of hypothalamic rhythm genes was detected by reverse transcription polymerase chain reaction (RT-PCR). Data revealed circadian loss of many neurotransmitters during the entire circadian cycle in the serum of group M, indicating that hypertension causes circadian rhythm disorders. Cosinor analysis of the rhythmic oscillations of the levels of 32 neurotransmitters in the three groups showed that the metabolite rhythms peaked at approximately the same time of day (ZT4 and ZT16). Moreover, the amplitudes of the metabolite rhythms were altered. After treatment with Uncaria, the amplitudes of 13 neurotransmitters reverted to normal. The change trends in the hypothalamic rhythm genes confirmed the rhythm changes in neurotransmitters. Collectively, a novel pharmacodynamic evaluation strategy was established to dynamically observe the holistic effects of Uncaria on 32 circulating neurotransmitters within 24 h from the perspective of time dimension.

Metabonomics study of essential hypertension and its chinese medicine subtypes by using gas chromatography-mass spectrometry and nuclear magnetic resonance spectroscopy.

A metabonomic study was performed to investigate the metabolic mechanism of essential hypertension and its Chinese medicine subtypes, including "Yin-deficiency and Yang-hyperactivity syndrome" (YDYHS) and "Yin-Yang deficiency syndrome" (YYDS). Plasma samples from 22 healthy volunteers, 31 hypertensive patients with YDYHS, and 29 hypertensive patients with YYDS were analyzed by (1)H-NMR spectroscopy and gas chromatography coupled with mass spectrometry (GC-MS). The three groups were distinctly classified by principal components analysis (PCA) and partial least squares discriminant analysis (PLS-DA). According to identified biomarkers and their related pathways, abnormal glucose metabolism might be the main common pathway from YDYHS to YYDS, and sympathetic nervous system activation would play an important role in the pathogenesis of YDYHS, while a low metabolic rate usually occurred in YYDS.

[Effects of rhynchophylla alkaloids on vascular adventitial fibroblast apoptosis and proliferation in the thoracic aorta of spontaneously hypertensive rats].

OBJECTIVE: To study the effects of rhynchophylline, isorhynchophylline, and rhynchophylla alkaloids on the vascular adventitial fibroblasts (VAF) apoptosis and proliferation in thoracic aorta of spontaneously hypertensive rats (SHR), and on the Bcl-2, Bax, c-Fos, c-Myc, laminin (LN), and fibronectin (FN). METHODS: Forty 8-week old male SHR were randomly divided into five groups, i. e., the model group, the captopril group (17.5 mg/kg), the isorhynchophylline group (5.0 mg/kg), the rhynchophylline group (5.0 mg/kg), and the rhynchophylla alkaloids group (50.0 mg/kg), 8 in each group. In addition, eight 8-week old male Wistar rats were selected as the normal group. Equal volume of normal saline was given to rats in the normal group and the model group by gastrogavage. Rats in the rest groups were perfused with isovolumic medication solution (10 mL/kg), six days per week for eight successive weeks. The dosage of drugs was adjusted according to the change of body weight. The VAF apoptosis rate of the thoracic aorta was measured by Annexin V-FITC combined with PI dyeing and flow cytometry. The protein expressions of thoracic aortic Bcl-2, Bax, c-Myc, c-Fos, FN, and LN were detected by immunohistochemical assay. The adventitial transforming growth factor beta1 (TGF-beta1) mRNA expression in the thoracic aorta was detected by in situ hybridization method. RESULTS: Compared with the model group, the tail arterial systolic pressure decreased, the VAF apoptosis and the protein expression of Bax increased, Bcl-2, c-Fos, FN, LN, and TGF-beta1 mRNA all decreased in the thoracic aorta of SHR in each treatment group after 4-and 8-week of intervention. Rhynchophylline, isorhynchophylline, and rhynchophylla alkaloids could inhibit the protein expression of c-Myc with statistical difference (P<0.05, P<0.01). Compared with the captopril group, there was no statistical difference in decreasing the tail arterial systolic pressure, the protein expression of c-Fos and the mRNA expression of TGF-beta1 among the rhynchophylline group, the isorhynchophylline group, and the rhynchophylla alkaloids group (P>0.05). There was statistical difference in increased VAF apoptosis and decreased protein expressions of Bcl-2, c-Myc, and LN (P<0.05, P<0.01). There was statistical difference in increased protein expression of Bax between the rhynchophylline group and the isorhynchophylline group (P<0.05, P<0.01). There was statistical difference in decreased protein expression of FN in the isorhynchophylline group (P<0.05). There was no significant difference among the rhynchophylline group, the isorhynchophylline group, or the rhynchophylla alkaloids group (P>0.05). CONCLUSIONS: Rhynchophylline, isorhynchophylline, and rhynchophylla alkaloids might promote the VAF apoptosis in the thoracic aorta of SHR by regulating the protein expressions of Bcl-2 and Bax. They might inhibit the VAF proliferation by restraining protein expressions of c-Fos, c-Myc, and TGF-beta1 mRNA. They also might improve the thoracic aorta wall reconstruction and decrease the tail arterial systolic pressure by down-regulating the protein expressions of FN and LN, and attenuating the deposition of extracellular matrix.

Aß damages learning and memory in Alzheimer's disease rats with kidney-yang deficiency.

Previous studies demonstrated that Alzheimer's disease was considered as the consequence produced by deficiency of Kidney essence. However, the mechanism underlying the symptoms also remains elusive. Here we report that spatial learning and memory, escape, and swimming capacities were damaged significantly in Kidney-yang deficiency rats. Indeed, both hippocampal Aß(40) and 42 increases in Kidney-yang deficiency contribute to the learning and memory impairments. Specifically, damage of synaptic plasticity is involved in the learning and memory impairment of Kidney-yang deficiency rats. We determined that the learning and memory damage in Kidney-yang deficiency due to synaptic plasticity impairment and increases of Aß(40) and 42 was not caused via NMDA receptor internalization induced by Aß increase. ß-Adrenergic receptor agonist can rescue the impaired long-term potential (LTP) in Kidney-yang rats. Taken together, our results suggest that spatial learning and memory inhibited in Kidney-yang deficiency might be induced by Aß increase and the decrease of ß(2) receptor function in glia.

[Determination of theanine in tea leaves by HPLC-inhibited chemiluminescence with Co2+ catalyst].

A method for the separation and determination of theanine in tea leaves has been developed, based on inhibited chemiluminescence of cobalt-catalyzed luminol-H2O2 reaction by theanine. Separations were performed on a YWG-C18 column using a mobile phase of 0.01 mol.L-1 sodium acetate-acetic acid buffer at pH 5.5 at a flow rate of 0.8 mL.min-1. The optimum concentrations of Co2+, luminol, and H2O2 were 2 micrograms.L-1, 0.25 mmol.L-1, and 0.5 mmol.L-1, respectively. Under the conditions mentioned above, the relative peak area (Y) of inhibited chemiluminescence of theanine was linear with the concentration (X) of theanine in the range of 0.2 g.L-1-5.0 g.L-1 and its linear regression equation was Y = 33,862 x + 1.0605 (r = 0.9983). The average recovery was 97.82% with a RSD of 2.16%.