Neuroprotective Effects of Ethanol Extract of Polyscias fruticosa (EEPF) against Glutamate-Mediated Neuronal Toxicity in HT22 Cells.

In traditional herbal medicine, the Polyscias fruticosa has been frequently used for the treatment of ischemia and inflammation. Oxidative stress mediated by elevated glutamate levels cause neuronal cell death in ischemia and various neurodegenerative diseases. However, so far, the neuroprotective effects of this plant extract against glutamate-mediated cell death have not been investigated in cell models. The current study investigates the neuroprotective effects of ethanol extracts of Polyscias fruticosa (EEPF) and elucidates the underlying molecular mechanisms of EEPFs relevant to neuroprotection against glutamate-mediated cell death. The oxidative stress-mediated cell death was induced by 5 mM glutamate treatment in HT22 cells. The cell viability was measured by a tetrazolium-based EZ-Cytox reagent and Calcein-AM fluorescent dye. Intracellular Ca2+ and ROS levels were measured by fluorescent dyes, fluo-3 AM and 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA), respectively. Protein expressions of p-AKT, BDNF, p-CREB, Bax, Bcl-2, and apoptosis-inducing factor (AIF) were determined by western blot analysis. The apoptotic cell death was measured by flow cytometry. The in vivo efficacy of EEPF was evaluated using the Mongolian gerbil mouse by surgery-induced brain ischemia. EEPF treatment showed a neuroprotective effect against glutamate-induced cell death. The EEPF co-treatment reduced the intracellular Ca2+ and ROS and apoptotic cell death. Furthermore, it recovered the p-AKT, p-CREB, BDNF, and Bcl-2 levels decreased by glutamate. The EEPF co-treatment suppressed the activation of apoptotic Bax, the nuclear translocation of AIF, and mitogen-activated protein kinase (MAPK) pathway proteins (ERK1/2, p38, JNK). Further, EEPF treatment significantly rescued the degenerative neurons in the ischemia-induced Mongolian gerbil in vivo model. EEPF exhibited neuroprotective properties that suppress glutamate-mediated neurotoxicity. The underlying mechanism of EEPF is increasing the level of p-AKT, p-CREB, BDNF, and Bcl-2 associated with cell survival. It has therapeutic potential for the treatment of glutamate-mediated neuropathology.

Kinetic Study on Chlorophyll and Antioxidant Activity from Polyscias fruticosa (L.) Harms Leaves via Microwave-Assisted Extraction.

Polyscias fruticosa (L.) leaves contain significant bioactive compounds with high antioxidant activity such as chlorophylls, total polyphenols, etc. but these have still been underutilized. In this study, the kinetics of chlorophyll and antioxidant activity extraction from P. fruticosa leaves by microwave-assisted extraction (MAE) were investigated. Microwave power was 300, 450, or 600 (W); the ratio of material/solvent varied from 1:40 to 1:80 (g/mL). In this study, the second-order kinetic model successfully predicted the change of chlorophyll and antioxidant activity during MAE. The increase of microwave power or/and the solvent amount increased saturated extraction efficiency and the extraction rate constant. However, the saturated concentration of chlorophyll and antioxidant activity increased with the increment of microwave power and the decrease in solvent amount.

Antioxidant and Anti-Inflammatory Mechanisms of Lipophilic Fractions from Polyscias fruticosa Leaves Based on Network Pharmacology, In Silico, and In Vitro Approaches.

Polyscias fruticosa leaf (PFL) has been used in food and traditional medicine for the treatment of rheumatism, ischemia, and neuralgia. However, the lipophilic components of PFL and their biological properties remain unknown. This study, integrating network pharmacology analysis with in silico and in vitro approaches, aimed to elucidate the antioxidant and anti-inflammatory capacities of lipophilic extracts from PFL. A total of 71 lipophilic compounds were identified in PFL using gas chromatography-mass spectrometry. Network pharmacology and molecular docking analyses showed that key active compounds, mainly phytosterols and sesquiterpenes, were responsible for regulating core target genes, such as PTGS2, TLR4, NFE2L2, PRKCD, KEAP1, NFKB1, NR1l2, PTGS1, AR, and CYP3A4, which were mostly enriched in oxidative stress and inflammation-related pathways. Furthermore, lipophilic extracts from PFL offered powerful antioxidant capacities, as evident in our cell-free antioxidant assays. These extracts also provided a protection against oxidative stress by inducing the expression of catalase and heme oxygenase-1 in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Additionally, lipophilic fractions from PFL showed anti-inflammatory potential in downregulating the level of pro-inflammatory factors in LPS-treated macrophages. Overall, these findings provide valuable insights into the antioxidant and anti-inflammatory properties of lipophilic extracts from PFL, which can be used as a fundamental basis for developing nutraceuticals and functional foods.

Growth and Oleanolic Acid Accumulation of Polyscias fruticosa Cell Suspension Cultures.

BACKGROUND: Oleanolic acid is an oleanane triterpene found in many plant species all over the world. This compound is also a major saponin in leaves of Polyscias fruticosa and possesses several promising pharmacological activities, such as hepatoprotective effects, and antiinflammatory, antioxidant, or anticancer activities. OBJECTIVE: The objective of the present work is to establish cell suspension culture of P. fruticosa, investigate the influence of several factors such as plant growth regulators and carbon source on cell growth, and determine their oleanolic acid content. METHODS: Cell culture was established by using 2 g fresh weight of 30 day old friable callus derived from in vitro stem segment in 50 mL of liquid medium with a shaking speed of 220 rpm. The culture was then incubated at 25±2ºC with a shaking speed of 120 rpm in the period of 12 h daylight at a light intensity of about 6.75 µmol/m2/s. Cell growth was measured by fresh and dry biomass at 16 h day. Oleanolic acid content was determined using HPLC analysis. RESULTS & DISCUSSION: The study results showed that MS medium containing 2% sucrose as a carbon source, supplemented with 1 mg/L 6-benzylaminopurine and 0.5 mg/L 2,4-dichlorophenoxyacetic acid was the most appropriate growth medium. Cell biomass and oleanolic acid content reached the highest values of 0.43 g dry weight/flask and 25.4 mg/g dry weight, respectively. CONCLUSION: These results indicated the potential production of oleanolic acid, a compound with high pharmacological value, from P. fruticosa cell culture.

Polysciosides J and K, two new oleanane-type triterpenoid saponins from the leaves of Polyscias fruticosa (L.) harms. cultivating in An Giang Province, Viet Nam.

For the first time, the phytochemical constituents of the leaves of Polyscias fruticosa (L.) Harms. cultivating in An Giang Province, Viet Nam were investigated and led to purify two new oleanane-type triterpenoid saponins, named polyscioside J (1) and polyscioside K (2) together with two known saponins, ladyginoside A (3) and chikusetsusaponin IVa (4) using variously chromatographic methods. Saponin (4) was reported for the first time from this species. Their structures were verified by IR, UV, HR-ESI-MS, NMR 1D and 2D experiments and compared with previous literatures.

Multiplication Coefficient of Polyscias fruticosa (L.) Harms under influence of the growth regulators

The study was carried out to examine the proliferation in vitro of Polyscias fruticosa (L.) Harms. Results: Using HgCl 2 with concentration of 0.1% within 15 minutes and H2O2 within 30 minutes was the best of decontamination time. With the time, the live rate was >80%. Meristems of the branch of Polyscias fruticosa (L.) Harms were induced to develop into plantlets in a medium containing MS-minerals and 2.0 mg/l kinetin. In this medium, an average of 3.4+-0.62 shoots in about 60 days. 100% rooting of the shoots were induced in MS medium supplemented with 1.0 mg/l IBA in 30 days. Rooted planted were grown in the green house.

In vitro propagation of Polyscias fruticosa (L.) Harms by direct regeneration from shoot tips

Study on preserved propagation of Polyscias fruticosa (L.) Harms – Araliaceae by direct regeneration from 3-years-old shoot tips. Shoot tips of 1.0 to 1.5cm long taken from field growing Polyscias fruticosa (L.) Harms were sterilized and cultured in Murashige and Skoog (MS) medium supplemented with 1.0mg/l BAP and 0.5mg/l IBA. After 8 weeks, these shoots began to regenerate directly from axil but quantity was not numerous and gave 2-3 new shoots after 4 more weeks. The results of rapid propagation of Polyscias fruticosa (L.) Harms’s shoots after 1, 3 and 4 months culture in MS + 2.0mg/l BAP + 0.5mg/l IBA environment showed that it was the best combination for shoot proliferation. The single shoots gained 2-3cm long were separated from shoot bushes and cultured in MS + 1.0mg/l IBA + 0.5mg/l BAP environment to make root the best. In this environment, shoots began to rooting after 11 days, gained 100% of rooting after 20 days. The plantlets were successfully transferred to PE bags. After 2 weeks, plantlets grew new roots with the rate of live 80-95%

Studies on Antioxidant Activity of Polyscias fruticosa Harms. (Araliaceae)

Extracts from roots and leaves of Polyscias fruticosa Harms. (Araliaceae) and their combination have been studied on lipid peroxidation in mouse brain using an auto-oxidation and a free radical generating system: the Fenton’s reagent (ferrous iron + hydrogen peroxide). All the root, leaf and combined extracts have inhibitory action on peroxidation of lipids and trolox, a water-soluble analogue of vitamin E used as a reference antioxidant compound. The results also indicate that the leaf extract is the strongest of all.

Studies on hepatoprotective activity of Polyscias fruticosa based on antioxidant mechanism

Experimentally, hepatoprotective effect of DinhLang was expressed in mice. The extract of Dinh Lang leaf suppressed the increase of MDA level in brain and in liver of mice suffered from CCl4- induced hepatitis with antioxydant activity stronger than that of mixed extract and extract prepared from the root. Dinh Lang had manifested liver protective effect in tetrachlorurcarbone –induced acute hepatitis. Antioxydant effect expressed in the inhibition against the processes of peroxydation of cell membrane can be one of the mechanism of liver protective effect of Dinh Lang