Network pharmacology identification and in Vivo validation of key pharmacological pathways of Phyllanthus reticulatus (Euphorbiaceae) leaf extract in liver cancer treatment.

ETHNOPHARMACOLOGICAL RELEVANCE: Phyllanthus reticulatus (Euphorbiaceae) is a medicinal plant that has been used in Zhuang medicine since ancient times. Traditionally, it has the effect of removing toxins and detumescence and can be used to treat hepatitis in China and India. Our previous studies have proved that the ethyl acetate extract of its leaves (PRPE) has an anti-hepatoma effect. AIM OF THE STUDY: To predict targets of an ethyl acetate extract of Phyllanthus reticulatus leaves (PRPE) in hepatoma treatment via network pharmacology and verify the predictions in a mouse model of liver cancer. MATERIALS AND METHODS: Chemical constituents and therapeutic targets of P. reticulatus (PRP) were searched and predicted via public databases. A protein-protein interaction network comprising common targets was constructed, and the key gene targets were identified. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used for biological function and pathway enrichment analyses. The effects of PRP on BEL-7404 and HepG2 cells were determined by MTT assay, apoptosis was measured by flow cytometry and hoechst44432/PI. And a nude mouse xenograft model was established to verify the anti-tumour effect in vivo. The histopathology of tumours was observed by staining with haematoxylin and eosin (H&E). Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were used to determine the gene and protein expression levels of phosphoinositide 3-kinase (PI3K), Akt1, p53, caspase-3, Bcl-2 and Bax, respectively. RESULTS: Twenty-seven chemical components and 567 potential therapeutic targets of PRP were identified. GO analysis indicated that these targets are mainly associated with peptidyl-tyrosine phosphorylation and steroid metabolic process. KEGG analysis showed that the targets are mainly located in the PI3K/Akt, apoptosis, mitogen-activated protein kinase (MAPK), Ras and vascular endothelial growth factor (VEGF) signalling pathways. According to the p-adjust value, the PI3K/Akt pathway is the core pathway. In vitro, PRPE could inhibit proliferation and induce apoptosis in hepatoma cells. IC50 values of PRPE were 2.48 and 6.34 mg/mL for BEL-7404 and hepG2 cells, respectively. PRPE significantly reduced tumour volume and weight. H&E results showed that PRPE repaired necrotic areas in hepatoma cells. PRPE reduced the protein expression of PI3K, Akt1 and Bcl-2 and increased the protein expression of p53 and Bax. Meanwhile, PRPE reduced the mRNA expression of PI3K, AKT1 and BCL2 and increased the mRNA expression of TP53, CASP3 and BAX. CONCLUSION: The targets of PRPE are the PI3K/Akt, apoptosis, MAPK, Ras and VEGF signalling pathways. Passing through the PI3K/Akt pathway to induce apoptosis is the main mechanism of PRPE.

Nematotoxic coumarins from Angelica pubescens Maxim. f. biserrata Shan et Yuan roots and their physiological effects on Bursaphelenchus xylophilus.

The ethanol extracts from the roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan was toxic against the pine wood nematode Bursaphelenchus xylophilus. The ethyl acetate-soluble fraction derived from this extract increased its potency with a mortality of 95.25% in 72 hr at 1.0 mg/mL. Four nematotoxic coumarins were obtained from the ethyl acetate extract by bioassay-guided isolation. These were identified as osthole 1, columbianadin 2, bergapten 3 and xanthotoxin 4 by mass and nuclear magnetic resonance spectral data analysis. The LC50 values against B. xylophilus in 72 hr were 489.17, 406.74, 430.08, and 435.66 µM, respectively. These compounds also altered the smooth morphology of the B. xylophilus exoskeleton to a rough and pitted appearance as visualized by electron microscopy. The coumarins 1-4 possessed significant acetylcholinesterase inhibitory activities but had negligible effects on amylase and cellulase. This research provides additional clues to the nematotoxic mechanism of coumarins against the pine wood nematode B. xylophilus. This work will assist in the development of coumarin nematicides with enhanced activity using molecular modifications of the core coumarin structure.The ethanol extracts from the roots of Angelica pubescens Maxim. f. biserrata Shan et Yuan was toxic against the pine wood nematode Bursaphelenchus xylophilus. The ethyl acetate-soluble fraction derived from this extract increased its potency with a mortality of 95.25% in 72 hr at 1.0 mg/mL. Four nematotoxic coumarins were obtained from the ethyl acetate extract by bioassay-guided isolation. These were identified as osthole 1, columbianadin 2, bergapten 3 and xanthotoxin 4 by mass and nuclear magnetic resonance spectral data analysis. The LC50 values against B. xylophilus in 72 hr were 489.17, 406.74, 430.08, and 435.66 µM, respectively. These compounds also altered the smooth morphology of the B. xylophilus exoskeleton to a rough and pitted appearance as visualized by electron microscopy. The coumarins 1-4 possessed significant acetylcholinesterase inhibitory activities but had negligible effects on amylase and cellulase. This research provides additional clues to the nematotoxic mechanism of coumarins against the pine wood nematode B. xylophilus. This work will assist in the development of coumarin nematicides with enhanced activity using molecular modifications of the core coumarin structure.

[Chemical constituents from Embelia laeta].

OBJECTIVE: To study the chemical constituents from Embelia laeta. METHODS: The constituents from the EtOAc fraction of ethanol extract of Embelia laeta were separated and purified by column chromatography with silica gel and polyamide. The compounds were identified by their physiochemical proerties and spectral data. RESULTS: Eleven compounds were isolated and identified as p-sitosterol (1), beta-daucosterel (2), gallic acid (3), vanillic acid (4), rutin (5), hyperin (6), quercetin (7), kaemperol ( 8), chrysoeriol (9), physcion(10) and apigenin-7-O-glucoside(ll). CONCLUSION: Compounds 5-11 are isolated from this genus for the first time.

[Studies on chemical constituents in n-butyl alcohol extract of roots of Actinidia deliciosa in Guangxi].

OBJECTIVE: To study the chemical constituents of n-butyl alcohol extract in the roots of Actinidia deliciosa in Guangxi. METHOD: The constituents were separated with various chromatographic techniques and their structures were elucidated by means of physicochemical properties and the analysis of their spectral data. RESULT: Six compounds were isolated and identified as eriantic acid B (1), 2alpha, 3beta, 24-trihydroxyursa-12-en-28-oic acid (2), 2alpha, 3alpha, 24-trihydroxyursa-12-en-28-oic acid (3), 2alpha, 3alpha, 23-tri-hydroxyursa-12, 20 (30)-dien-28-oic acid (4), 2alpha, 3alpha, 24-trihydroxyursa-12, 20 (30)-dien-28-oic acid (5), n-butyl-O-beta-D-fruto-pyranoside (6). CONCLUSION: Compounds 1-4, 6 were obtained from this plant for the first time. Compound 6 was obtained from the genus Actinidia for the first time.

[Studies on the chemical constituents of the ethyl acetate portion of Nervilia fordii].

OBJECTIVE: To study the chemical constituents of the ethyl acetate portion in the herb of Nervilia fordii from guangxi. METHODS: The constituents were separated and purified by using column chromatography with silica gel. These compounds were identified by their physical and spectral data. RESULTS: Five compounds were isolated and identified as norleucine (crystal I), 24 (S/beta)-dihydrocycloeucalenol-(E)-p-hydroxy cinnamate (crystal II) , rhamnocitrin (crystal III), rhamnazin (crystal IV), daucosterol (crystal V). CONCLUSION: Compounds I , II, III, IV, V were isolated from this plant for the first time.

[Study on anticancer effect in vivo of active fraction from Nervilia fordii].

OBJECTIVE: To determine the active fraction with anticancer effect in vivo from Nervilia fordii. METHODS: The effective petroleum ether extract and ethyl acetate extract parts preliminary were selected in vitro, then anticancer experiments in vivo were done by S180-mice and H22-mice models. RESULTS: Petroleum ether extract and ethyl acetate extract parts both had obvious anticancer effects to S180-mice and H22-mice, and could prolong H22-mice life. Meanwhile, they could improve the immunoloregulation of mice. CONCLUSION: It is the first time that the petroleum ether extract and ethyl acetate extract of Nervilia foadii are proved to be the effective anticancer fractions in vivo. On this basis, the further studies are needed on active principles or principle group with anticancer effect and the characteristics of this effect in Nervilia foadii.