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Identifying Active Compounds and Mechanism of Camellia nitidissima Chi on Anti-Colon Cancer by Network Pharmacology and Experimental Validation.

ABSTRACT: Camellia nitidissima Chi (CNC) is a traditional Chinese medicine (TCM) with anticancer property. However, its underlying mechanisms of anti-colon cancer (CC) remain unknown. Therefore, a systematic approach is proposed in the present study to elucidate the anticancer mechanisms of CNC based on network pharmacology and experimental validation. Initially, the potential active ingredients of CNC were verified via the TCMSP database based on the oral bioavailability (OB) and drug-likeness (DL) terms. Hub targets of CNC were acquired from SwissTarget prediction and TCMSP databases, and target genes related to CC were gathered from GeneCards and OMIM databases. Cytoscape was used to establish the compound-target networks. Next, the hub target genes collected from the CNC and CC were parsed via GO and KEGG analysis. Results of GO and KEGG analysis reveal that quercetin and luteolin in CNC, VEGFA and AKT1 targets, and PI3K-Akt pathway were associated with the suppression of CC. Besides, the result of molecular docking unveils that VEGFA demonstrates the most powerful binding affinity among the binding outcomes. This finding was successfully validated using in vitro HCT116 cell model experiment. In conclusion, this study proved the usefulness of integrating network pharmacology with in vitro experiments in the elucidation of underlying molecular mechanisms of TCM.

SUBMITTER: Chen Y

PROVIDER: S-EPMC8413042 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Proteome Analysis of Camellia nitidissima Chi Revealed Its Role in Colon Cancer Through the Apoptosis and Ferroptosis Pathway.

Project description:Colon cancer is the third most common cancer in the world with a high mortality rate. At present, surgery combined with radiotherapy and chemotherapy is the primary treatment, but patient prognosis remains poor. Traditional Chinese medicine (TCM) has become a complementary and alternative source of anti-cancer drugs. Camellia nitidissima Chi (CNC) is a TCM used to treat a variety of cancers. However, the role of CNC in cancer remains elusive, and its effect and mechanism on colon cancer have not been reported. Here, we show that CNC exerts an excellent inhibitory effect on colon cancer proliferation and apoptosis induction in vitro and in vivo. We performed label free-based quantitative proteomic analysis to evaluate the HCT116 cells treated with CNC. Our data revealed a total of 363 differentially expressed proteins, of which 157 were up-regulated and 206 down-regulated. Gene Ontology enrichment analysis showed that these proteins were involved in tumor occurrence and development through multiple biological processes such as cell proliferation, cell apoptosis, cell cycle, and cell death. Interestingly, we also found significant changes in ferroptosis pathways. The role of essential proteins glutathione peroxidase 4 (GPX4) and heme oxygenase-1 (HMOX1) were verified. CNC decreased the expression of GPX4 and increased the expression of HMOX1 at the mRNA and protein levels in vivo and in vitro. Collectively, these findings reveal that CNC regulates colon cancer progression via the ferroptosis pathway and could be an attractive treatment for colon cancer.

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Novel Graphene/In2O3 Nanocubes Preparation and Selective Electrochemical Detection for L-Lysine of Camellia nitidissima Chi.

Project description:In this work, novel graphene/In2O3 (GR/In2O3) nanocubes were prepared via one-pot solvothermal treatment, reduction reaction, and successive annealing technology at 600 °C step by step. Interestingly, In2O3 with featured cubic morphology was observed to grow on multi-layered graphene nanosheets, forming novel GR/In2O3 nanocubes. The resulting nanocomposites were characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction spectroscopy (XRD), etc. Further investigations demonstrated that a selective electrochemical sensor based on the prepared GR/In2O3 nanocubes can be achieved. By using the prepared GR/In2O3-based electrochemical sensor, the enantioselective and chem-selective performance, as well as the optimal conditions for L-Lysine detection in Camellia nitidissima Chi, were evaluated. The experimental results revealed that the GR/In2O3 nanocube-based electrochemical sensor showed good chiral recognition features for L-lysine in Camellia nitidissima Chi with a linear range of 0.23-30 ?mol·L-1, together with selectivity and anti-interference properties for other different amino acids in Camellia nitidissima Chi.

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Antioxidant capacity of phenolics in Camellia nitidissima Chi flowers and their identification by HPLC Triple TOF MS/MS.

Project description:Camellia nitidissima Chi (CNC) is a valuable medicinal and edible plant in China. In this study, CNC flowers were extracted with 95% ethanol, then partitioned into dichloromethane, ethyl acetate, n-butanol, and water fractions, with the antioxidant capacity of flavonoids and other phytochemicals in CNC flowers investigated for the first time. Results showed that the ethyl acetate fraction exhibited the strongest antioxidant capacity and highest total phenolic content (TPC) compared with the other fractions. Furthermore, in the ethyl acetate fraction, the 50% effective concentrations (EC50) of ABTS+ and DPPH radical scavenging activities were 64.24 ± 1.80 and 78.80 ± 0.34 μg/mL, respectively, and the ferric reducing antioxidant power (FRAP) was 801.49 ± 2.30 μM FeSO4 at 1,000 μg/mL. Pearson's correlation coefficients and principal component analyses (PCA) for the TPC and antioxidant capacity of the five fractions indicated that the phenolic compounds were the major antioxidant constituents in the flowers. To exploit the antioxidants in CNC flowers, 21 phenolic compounds in the ethanolic extract fraction were identified by HPLC Triple TOF MS/MS, next, 12 flavonoids were isolated and elucidated, of which compounds 1-5 showed potent antioxidant capacity. In addition, the potential structure-activity relationship among these 12 flavonoids showed that (1) the o-catechol group in the B-ring was primarily responsible for the antioxidant capacity of flavonoids and (2) steric hindrance, produced by glycosides and other groups, could reduce the antioxidant capacity of the flavonoids.

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Linear Graphene Nanocomposite Synthesis and an Analytical Application for the Amino Acid Detection of Camellia nitidissima Chi Seeds.

Project description:Husk derived amino modified linear graphene nanocomposites (aLGN) with a diameter range of 80-300 nm and a length range of 100-300 μm were prepared by a modified Hummers method, ammonia treatment, NaBH₄ reduction and phenylalanine induced assembly processes, etc. The resulting composites were characterized by transmission electron microscopy (TEM), atomic force microscopy (AFM), scanning electron microscopy (SEM), biological microscope (BM), and X-ray diffraction spectroscopy (XRD), etc. Investigations found that the aLGN can serve as the novel coating of stir bar sorptive extraction (SBSE) technology. By combing this technology with gas chromatography-mass spectrometry (GC-MS), the combined SBSE/GC-MS technology with an aLGN coating can detect seventeen kinds of amino acids of Camellia nitidissima Chi seeds, including Ala, Gly, Thr, Ser, Val, Leu, Ile, Cys, Pro, Met, Asp, Phe, Glu, Lys, Tyr, His, and Arg. Compared to a conventional polydimethylsiloxane (PDMS) coating, an aLGN coating for SBSE exhibited a better thermal desorption performance, better analytes fragmentation depressing efficiencies, higher peak intensities, and superior amino acid discrimination, leading to a practicable and highly distinguishable method for the variable amino acid detection of Camellia nitidissima Chi seeds.

| S-EPMC5506894 | biostudies-literature

Functional Diversification of the Dihydroflavonol 4-Reductase from Camellia nitidissima Chi. in the Control of Polyphenol Biosynthesis.

Project description:Plant secondary metabolism is complex in its diverse chemical composition and dynamic regulation of biosynthesis. How the functional diversification of enzymes contributes to the diversity is largely unknown. In the flavonoids pathway, dihydroflavonol 4-reductase (DFR) is a key enzyme mediating dihydroflavanol into anthocyanins biosynthesis. Here, the DFR homolog was identified from Camellia nitidissima Chi. (CnDFR) which is a unique species of the genus Camellia with golden yellow petals. Sequence analysis showed that CnDFR possessed not only conserved catalytic domains, but also some amino acids peculiar to Camellia species. Gene expression analysis revealed that CnDFR was expressed in all tissues and the expression of CnDFR was positively correlated with polyphenols but negatively with yellow coloration. The subcellular localization of CnDFR by the tobacco infiltration assay showed a likely dual localization in the nucleus and cell membrane. Furthermore, overexpression transgenic lines were generated in tobacco to understand the molecular function of CnDFR. The analyses of metabolites suggested that ectopic expression of CnDFR enhanced the biosynthesis of polyphenols, while no accumulation of anthocyanins was detected. These results indicate a functional diversification of the reductase activities in Camellia plants and provide molecular insights into the regulation of floral color.

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Neuroprotective effects of Camellia nitidissima Chi leaf extract in hydrogen peroxide-treated human neuroblastoma cells and its molecule mechanisms.

Project description:Camellia nitidissima Chi (CNC) is a famous medicinal and edible plant with the name of "Tea for Longevity" in Guangxi province of China. In present study, we determined the protective effect of extract from CNC leaves on H2O2-induced cell injury and its underlying mechanisms in human neuroblastoma (SH-SY5Y) cells. The ethyl acetate fraction of CNC leaves (CLE, 50-200 μg/ml) treatment significantly increased the cell viability of H2O2-treated SH-SY5Y cells and reduced the leakage of LDH in a reversed "U"-shape manner. It was confirmed by Hoechst 33,342 staining that CLE attenuated H2O2-induced apoptosis in SH-SY5Y cells. The CLE (100 and 150 μg/ml) treatment significantly relieved H2O2-induced oxidative stress by decreasing intracellular ROS level, and increasing the activities of superoxide dismutase (SOD) and catalase (CAT). Western blot analysis demonstrated that the CLE treatment reserved H2O2-induced decrease of pCREB (Ser133) expression, and its downstream protein BDNF. In addition, 37 phenolic compounds in CLE were identified by UPLC-TOF MS/MS, and the main active phytochemicals seemed to be catechins, quercetin, kaempferol, and their derivatives. In conclusion, the data analysis showed that the neuroprotective effect of CNC leaves might be achieved via synergistically boosting endogenous antioxidant defenses and neurotrophic signaling pathway. These results suggest that CNC leaves are valuable resources for functional foods and beverages.

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Nuclear Magnetic Resonance-Based Metabolomics Approach to Evaluate the Prevention Effect of Camellia nitidissima Chi on Colitis-Associated Carcinogenesis.

Project description:Colorectal cancer (CRC) is one of the most common malignant tumors worldwide, occurring in the colon or rectum portion of large intestine. With marked antioxidant, anti-inflammation and anti-tumor activities, Camellia nitidissima Chi has been used as an effective treatment of cancer. The azoxymethane/dextran sodium sulfate (AOM/DSS) induced CRC mice model was established and the prevention effect of C. nitidissima Chi extracts on the evolving of CRC was evaluated by examination of neoplastic lesions, histopathological inspection, serum biochemistry analysis, combined with nuclear magnetic resonance (NMR)-based metabolomics and correlation network analysis. C. nitidissima Chi extracts could significantly inhibit AOM/DSS induced CRC, relieve the colonic pathology of inflammation and ameliorate the serum biochemistry, and could significantly reverse the disturbed metabolic profiling toward the normal state. Moreover, the butanol fraction showed a better efficacy than the water-soluble fraction of C. nitidissima Chi. Further development of C. nitidissima Chi extracts as a potent CRC inhibitor was warranted.

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Project description:Camellia nitidissima cultivar:Camellia nitidissima Transcriptome or Gene expression

| PRJNA392948 | ENA

Active fractions of golden-flowered tea (Camellia nitidissima Chi) inhibit epidermal growth factor receptor mutated non-small cell lung cancer via multiple pathways and targets in vitro and in vivo

Project description:As a medicine-food homology (MFH) plant, golden-flowered tea (Camellia nitidissima Chi, CNC) has many different pharmacologic activities and is known as “the queen of the tea family” and “the Panda of the Plant world”. Several studies have revealed the pharmacologic effects of CNC crude extract, including anti-tumor, anti-oxidative and hepatoprotective activity. However, there are few studies on the anti-tumor active fractions and components of CNC, yet the underlying mechanism has not been investigated. Thus, we sought to verify the anti-non-small cell lung cancer (NSCLC) effects of four active fractions of CNC. Firstly, we determined the pharmacodynamic material basis of the four active fractions of CNC (Camellia. leave. saponins, Camellia. leave. polyphenols, Camellia. flower. saponins, Camellia. flower. polyphenols) by UPLC-Q-TOF-MS/MS and confirmed the differences in their specific compound contents. Then, MTT, colony formation assay and EdU incorporation assay confirmed that all fractions of CNC exhibit significant inhibitory on NSCLC, especially the Camellia. leave. saponins (CLS) fraction on EGFR mutated NSCLC cell lines. Moreover, transcriptome analysis revealed that the inhibition of NSCLC cell growth by CLS may be via three pathways, including “Cytokine-cytokine receptor interaction,” “PI3K-Akt signaling pathway” and “MAPK signaling pathway.” Subsequently, quantitative real-time PCR (RT-qPCR) and Western blot (WB) revealed TGFB2, INHBB, PIK3R3, ITGB8, TrkB and CACNA1D as the critical targets for the anti-tumor effects of CLS in vitro. Finally, the xenograft models confirmed that CLS treatment effectively suppressed tumor growth, and the key targets were also verified in vivo. These observations suggest that golden-flowered tea could be developed as a functional tea drink with anti-cancer ability, providing an essential molecular mechanism foundation for MFH medicine treating NSCLC.

| S-EPMC9649924 | biostudies-literature

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Project description:Camellia nitidissima Raw sequence reads

| PRJNA899601 | ENA