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OBJECTIVE To study the inhibitory effect and mechanism of total flavonoids from Melicope pteleifolia （TF-MPL） on transplanted tumor of colorectal cancer in nude mice. METHODS The transplanted tumor model of colorectal cancer was induced by injecting 0.2 mL colorectal cancer cell LoVo subcutaneously via the right armpit of nude mice. After successful modeling， nude mice were randomly divided into model group， 5-fluorouracil group （positive control， 10 mg/kg）， TF-MPL high- dose and low-dose groups （25， 12.5 mg/kg）； a normal group （normal saline containing 0.3% carboxymethyl cellulose sodium） without modeling was additionally set up， with 6 mice in each group. Each group was intraperitoneally injected with the corresponding drug solution/solvent for 21 consecutive days. The inhibitory rate of the transplanted tumor， liver and spleen index， and the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum were detected after the last medication； the morphological changes of tumor tissue were observed； immunohistochemical staining was used to detect protein expressions of Toll- like receptor 4 （TLR4） and nuclear factor-κB subunit p65 （NF-κB p65） in tumor tissue of nude mice. Western blot assay was used to detect protein expressions of TLR4， myeloid differentiation factor 88 （MyD88）， TNF receptor-associated factor 6 （TRAF6）， interleukin-1 receptor-associated kinase 1 （IRAK-1）， NF-κB p65 and caspase-3 in tumor tissue of nude mice. RESULTS Compared with the model group， TF-MPL high-dose group showed a significant decrease in tumor weight （inhibitory rate of 36.91%）， liver and spleen index， serum levels of TNF-α and IL-6 and protein expressions of TLR4， MyD88， TRAF6，IRAK-1 and NF- κB p65 （P＜0.05 or P＜0.01）； the expression of caspase-3 protein was increased significantly （P＜0.05）， and more tumor cell shrinkage and deformation， nuclear pyknosis and fragmentation were observed. CONCLUSIONS TF-MPL can significantly inhibit the growth of transplanted tumor of colorectal cancer in nude mice， the mechanism of which may be associated with reducing inflammatory response， inhibiting TLR4/MyD88/NF-κB signaling pathway， and promoting apoptosis in colorectal cancer cells.
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Objective: To obtain the transcriptome sequence database of Melicope pteleifolia. Methods: The transcriptome sequencing and systematic bioinformatics analysis were carried out using the second generation high-throughput sequencing platform Illumina HiSeqTM 2000 with mixed root, stem and leaf samples of M. pteleifolia. Results: A total of 47 045 040 high quality sequences (clean reads) were obtained by transcriptome sequencing analysis. A total of 67 956 unigenes were assembled by Trinity de novo, with an average length of 787 nt. BLAST analysis showed that 42 749 (61.92%), 31 152 (45.84%), 26 563 (39.09%), and 17 481 (25.72%) unigenes were annotated in NR, Swiss port, KOG and KEGG databases respectively, and 47 groups were involved in three GO classification: biological process, cellular component and molecular function. A total of 9807 unigenes were annotated to 130 KEGG metabolic pathways, 19 secondary metabolic pathways were screened. Twenty-five different KOG functional groups were obtained by the analysis of KOG functional classification. It was predicted that there were 56 families of higher plant transcription factors. A total of 7 748 simple sequence repeats (SSRs) were found by MISA software. The number (4 117) of the tri-nucleotide SSRs was the richest, with a frequency of 53.1%, and the number of the penta-nucleotide SSRs was relatively small, accounting for 2.2%. Conclusion: The transcriptome information characteristics of root, stem, and leaf of M. pteleifolia can be obtained by high-throughput Illumina sequencing technology and bioinformatics analysis, which will lay a foundation for further research on functional gene mining, secondary metabolic pathway analysis and regulation mechanism of M. pteleifolia.
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Melicope pteleifolia belongs to the family Rutaceae, Sect. Tetradium (Lour.) Engl. plants, which is widely distributed in Guangxi, Guangdong, Hainan, Jiangxi, Fujian, Guizhu, Yunnan, and other provinces. The main components are volatile oils and alkaloids. M. pteleifolia has been reported that it has antipyretic, analgesic, anti-inflammatory effects, and so on. In this paper, the investigation of documents, chemical components, and pharmacological effects is reviewed, which can provide a theoretical basis for further research of M. pteleifolia.
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Objective To study the constituents in Melicope pteleifolia. Methods Plant material was isolated with 80% EtOH. Compounds were separated with chromatographic methods and their structures were elucidated on the basis of spectral analysis (EI-MS, 1H-NMR, and 13C-NMR) and chemical evidence. Results Five compounds were isolated from petrol ether or ethyl acetate soluble fraction. Their structures were identified as 3,5,3'-trihydroxy-4'-methoxy-7-(3-methylbut-2-enyloxy) flavone (pteleifolosin C, 1), 3,7-dimethoxyl kaempferol (kamatakenin, 2), vanillic acid (3), tricosanoic acid tetradecyl ester (4), and p-sitosterol (5), respectively. Conclusion Compound 1 is a new structure named pteleifolosin C. Compounds 2-4 are isolated from this plant for the first time.