Pinus Morrisonicola Research Articles

Anti-Inflammatory Effect of Medicinal Fungus Antrodia cinnamomea Cultivated on Pinus morrisonicola Hayata.

The fungus Antrodia cinnamomea, which grows on Cinnamomum kanehirae tree, has many medicinal uses. However, its cultivation using the traditional method of growing on the C. kanehirae tree is costly and time-consuming. A possible alternative method of cultivating A. cinnamomea is to use Pinus morrisonicola Hayata tree, as it contains α-terpineol, which stimulates the synthesis of triterpenoids. To compare the cultivation of A. cinnamomea on P. morrisonicola and C. kanehirae, the contents of triterpenoids and antcin were determined using high-performance liquid chromatography. Anti-inflammatory effects of the extracts of each product were investigated in lipopolysaccharide (LPS)-stimulated BV-2 cells. Their mechanisms on mitogen-activated protein kinase (MAPK) signalling pathways (p38, c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK)) were determined using Western blot analysis. The results showed that the cultivation times of A. cinnamomea on P. morrisonicola and traditional C. kanehirae discs were drastically different, lasting 6 and 18 months, respectively. The concentration of triterpenoids in the corresponding fruiting bodies was (70.0±3.0) and (20.0±4.0) mg/mL, respectively. More antcins were produced in the P. morrisonicola culture. Similar anti-inflammatory effect was obtained by both cultures, which is confirmed by the reduced production of IL-1β, IL-6, COX-2 and nitrogen monoxide. Their mechanisms were confirmed by the suppression of MAPK signalling pathways. Cultivation on P. morrisonicola is an innovative and more cost-effective method for growing A. cinnamomea. The same anti-inflammatory effect is achieved in a shorter production time.

Principle active metabolites of Pinus morrisonicola Hayata synergistically inhibit cell proliferation and autophagy to elevate apoptosis in hepatocellular carcinoma cells.

Hepatocellular carcinoma (HCC), a common primary tumor of liver is a leading cause of cancer-associated deaths. Improving cellular apoptosis and enhancing autophagic clearance is been considered to improve treatment outcomes of HCC. Polyphenols from Pinus morrisonicola (Hayata) have shown various physiological and therapeutic benefits and the flavonoid chrysin is been known for their anticancer effects. However, the main bioactive principle and the mechanism underlying the antitumor activity of pine needle extract are not clear yet. In this study, the effects of ethanol extract from pine needle on HCC cells were determined. The results show that when compared with administration of chrysin alone, a fraction containing pinocembrin, chrysin, and tiliroside significantly reduced autophagy and increased apoptosis. The results also correlated with decrease in cell cycle regulators and the autophagic proteins like LC3-II. Collectively, the results imply the fraction containing pinocembrin, chrysin, and tiliroside as an ideal complementary medicine for an effective antitumor activity.

Anti-Aging Constituents from Pinus morrisonicola Leaves.

Pinus morrisonicola Hayata is a unique plant species found in Taiwan. Previous studies have identified its anti-hypertensive, anti-oxidative, and anti-inflammatory effects. In this study, a bioactivity-guided approach was employed to extract 20 compounds from the ethyl acetate fraction of the ethanol extract of Pinus morrisonicola Hayata's pine needles. The anti-aging effects of these compounds were investigated using HT-1080 cells. The structures of the purified compounds were confirmed through NMR and LC-MS analysis, revealing the presence of nine flavonoids, two lignans, one coumarin, one benzofuran, one phenylic acid, and six diterpenoids. Among them, PML18, PML19, and PML20 were identified as novel diterpene. Compounds 3, 4, and 5 exhibited remarkable inhibitory effects against MMP-2 and showed no significant cell toxicity at 25 μM. Although the purified compounds showed lower activity against Pro MMP-2 and Pro MMP-9 compared to the ethyl acetate fraction, we speculate that this is the result of synergistic effects.

In Vivo Hypolipidemic Effects and Antioxidant Capacity of Pinus morrisonicola Hay Extracts by Supercritical Carbon Dioxide Extraction

Pinus morrisonicola hay (PM) is a pine tree unique to Taiwan, whose needles are used as traditional medicine and as functional drink. PME3-1 was made using supercritical extraction to evaluate the prevention of hyperlipidemia. This study explored the hypolipidemic effect of PME3-1 on hamsters on a high fat and cholesterol (HFC) diet. Three groups of hamsters were fed with PME3-1 (0.2, 1.0, and 5.0 mg/kg bw). After feeding for eight weeks, PME3-1 reduced the serum cholesterol, triglyceride levels, the low-density lipoprotein cholesterol/high-density lipoprotein cholesterol (LDL/HDL) ratio, and the swelling of the liver and kidney significantly (p < 0.05). In addition, feeding the hamsters with 5.0 mg/kg bw of PME3-1 could significantly reduce their total lipid (TL) content, total cholesterol (TC) content, total triglyceride (TG) content, and the HMG-CoA reductase activity in the liver (p < 0.05). Meanwhile, the antioxidant enzymes in the liver, glutathione peroxidase (GPx) and glutathione S-transferase (GST), can also improve, promoting the excretion of triglycerides (TG) and total cholesterol (TC) in the feces (p < 0.05). Therefore, these results confirm that PME3-1 hypolipidemic and antioxidant regulating functions in vivo.

Composition analysis of exudates produced by conifers grown in Taiwan and their antifungal activity

Exudates are involved in the defense mechanism of trees; they could work against insects or microorganisms through a physical or chemical system. The main components of exudates are terpenoids. This study identified the main compounds of exudates from 13 conifers of Taiwan using gas chromatogram–mass spectrometry (GC–MS) and spectroscopic analysis. The results revealed that the main volatiles were α-pinene, β-ocimene, β-pinene, sabinene, and caryophyllene. On the other hand, the main nonvolatile compounds were diterpenoids, which were classified into three skeletons (abietane-, labdane-, and pimarane-types). Among these, abietane-type presented in Pinaceae and in most of Cupressaceae; labdane-type presented in Pinaceae and in all of Cupressaceae and Araucariaceae; pimarane-type existed in both Pinaceae and Cupressaceae. Furthermore, the epigenetics of conifers analysis results by GC–MS and heteronuclear single quantum coherence (HSQC) of nuclear magnetic resonance (NMR) fingerprints were similar to traditional taxonomy classification; it indicated that exudates chemotaxonomy by using GC–MS and HSQC profiling is a useful technology to classify the conifers. Besides, the exudates of Pinus elliottii, Pinus taiwanensis, Calocedrus macrolepis and Chamaecyparis formosensis possessed the strong antifungal activity. For white-rot fungus, Trametes versicolor, Pinus morrisonicola, Chamaecyparis obtusa, and Araucaria heterophylla exhibited the higher antifungal index. For brown-rot fungus, Laetiporus sulphureus, Pinus elliottii, Pinus morrisonicola, and Chamaecyparis formosensis revealed a good antifungal activity.

Pinus morrisonicola

This datasheet on Pinus morrisonicola covers Identity.

Unraveling microbiomes associated with decomposition of needles of two Pinus species with contrasting fire-adaptive strategies

Pines evolve with fire and have diverged into fire-avoiders and fire-adapters. Pine needles are resistant to microbial decomposition, and their accumulation serves as a major fuel source for forest fires. Facilitated needle decomposition reduces forest fuel loads and probably interacts with the adaptive strategies of pines. However, there is little information available regarding differences in microbiomes associated with needle decomposition of the two types of pine. In this study, we monitored the microbial communities in three compartments associated with needle decomposition, namely, fresh needles, needle litter, and humus, along altitudinal gradients at three sites of the fire-tolerator Pinus taiwanensis and one site of the fire-avoider Pinus morrisonicola in Taiwan. The humus microbiome was dominated by an overwhelming proportion of bacteria. As a reservoir of decomposers, humus bacteria accounted for > 70% of the litter communities and showed marked differences in composition between the two Pinus species, as determined by non-metric multidimensional scaling. Such differences were associated with the abundance of bacterial genes encoding lignin-degrading enzymes, which were highest in the P. morrisonicola stand. Additionally, the humus accumulating under this fire-avoider was characterized by low ammonium concentrations and lower pH compared with that under the fire-tolerator, which is indicative of conditions conducive to litter decomposition. Conclusively, our findings suggested that P. morrisonicola grows in more favorable conditions for needle decomposition compared with the habitats where the P. taiwanensis lives.

Pinus morrisonicola needles essential oil nanoemulsions as a novel strong antioxidant and anticancer agent

Regarding the anticancer effects of pine (Pinus morrisonicola, PM) needle essential oil (PNEO) and its rich bioactive contents we aimed to improve PNEO bio accessibility and efficiency by nano emulsification and evaluate its biological effects. For this purpose, ultrasound nano emulsification was utilized to synthesis of PNEO Nano emulsions (PNEO-NE). Then the PNEO-NE was characterized and evaluated its biological properties including cytotoxic, antioxidant and pro apoptotic effects by cellular and molecular methods. The results confirmed the formation of PNEO Nano emulsion with an average diameter of 41.16 nm and dispersion index of 0.31. The PNEO-NE showed a greater inhibitory effect against cancer cells compared to normal cell (HFF). Moreover, the significantly up-regulated genes (Cas-3, Cas-9, VEGF/VEGFR, CAT and SOD) and increased subG1 peaks indicate the apoptotic death in HT-29 cells. PNEO-NE inhibited ABTS and DPPH free radicals (IC50 4 and 40 µg/ml). According to results, the safe efficient PNEO-NE system displays a notable cancer cell-sensitive cytotoxic impact by inducing apoptotic mediated pathways.

Anti-inflammatory properties of fermented pine (Pinus morrisonicola Hay.) needle on lipopolysaccharide-induced inflammation in RAW 264.7 macrophage cells.

This study was aimed to explore the antioxidant and anti-inflammatory activity of various pine needle products (non-fermented, fermented, and commercial) extracted with different solvents (hexane, ethyl acetate, and water) in lipopolysaccharide (LPS) induced RAW 264.7 cells. The phenolic/flavonoid contents of ethyl acetate extract of fermented pine needle (EAE-FPN) is higher than other pine product extracts (hexane/water). The levels of antioxidant indices (TEAC, DPPH) as well as free radical scavenging activity (H2 O2 ) were significantly improved in EAE-FPN than other pine needle product extracts. The levels of ROS and various inflammatory markers (NO, PGE2, TNF-α, and IL-1β/6) were considerably abolished by EAE-FPN in a dose-dependent manner (50-200μg/ml). Moreover, the protein expressions of inducible NO synthase (iNOS), cyclooxygenase 2 (COX-2), and nuclear factor Kappa B (NF-κB) p65 subunit were also markedly downregulated by EAE-FPN. Collectively, EAE-FPN with phenolic/flavonoid content showed excellent antioxidant and anti-inflammatory properties via modulating NF-κB signaling pathway. PRACTICAL APPLICATIONS: Pine needle drink (Pinus morrisonicola Hay) has been used as a functional beverage for many years due to its various biological properties in Asia especially in Taiwan, China, and Korea. Many researchers hinted various biological activity of fermented pine needle product but none of them explored the in-depth mechanism underpinning its antioxidant and anti-inflammatory properties in LPS-induced RAW 264.7 cell model. Hence, the current cell line study was designed to assess the underlying mechanism behind the antioxidant and anti-inflammatory activity of Pine needles extract (both non-fermented and fermented) in LPS-induced RAW 264.7 cells (macrophage). The outcome of this study distinctly showed that EAE-FPN displayed potent antioxidant and anti-inflammatory activities by regulating NF-κB signaling pathway. Therefore, pine needle could be developed into functional drink to abolish the progression of inflammatory responses in various disease condition.

Characterization of the complete chloroplast genome of Pinus wangii (Pinaceae), an endangered and endemic species in China

The complete cp genome of Piuns wangii was 117,014 bp in size, containing a large single copy (LSC, 64,061 bp) region and a small single copy (SSC, 52,007 bp) region separated by two reduced inverted repeats (IRs, 473 bp). A total of 107 genes were predicted, including 73 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Most of the 51 simple sequence repeats (SSRs) were mononucleotides motifs of A/T types and were found to be located in non-coding regions. Phylogenetic analysis of Asian Pinus showed P. wangii clustered a strongly-supported clade with Pinus morrisonicola, Pinus sibirica, Pinus wallichiana, and Pinus pumila.

Characterization of Vasorelaxant Principles from the Needles of Pinus morrisonicola Hayata.

Pinus morrisonicola Hayata, usually called Taiwan five-leaf pine (5LP), is an endemic species in Taiwan and is traditionally used to relieve hypertension symptoms and improve cardiovascular function. In this study, the needle extract of 5LP was fractionated and analyzed by LC/MS/MS to search for possible antihypertensive candidates. In addition, bioassay-guided purification of the bioactive components was performed by Ca2+ fluorescent signal (Fluo 4-AM) assays. Two dihydrobenzofuran lignans, pinumorrisonide A (1) and icariside E4 (2), and one acylated flavonoid glycoside, kaempferol 3-O-α-(6‴-p-coumaroylglucosyl-β-1,4-rhamnoside) (3) were characterized from the active fractions. The structure of a new compound 1 was established on the basis of 2D NMR spectroscopic and mass spectrometric analyses, and the known compounds 2 and 3 were identified by comparison of their physical and spectroscopic data with those reported in the literature. The purified compounds 1–3 exhibited significant inhibition of Ca2+ fluorescence with IC50 values of 0.71, 0.36, and 0.20 mM, respectively. A mechanism study showed that these compounds showed vasorelaxant effects by blocking the voltage-operated Ca2+ channel (VOCC) and inhibiting Ca2+ influx to the cytoplasmic. These results suggested that 5LP and the three characterized components could be promising antihypertensive candidates for the use as VOCC blockers.

Extracting antioxidant phenolic compounds from compressional-puffing pretreated Pinus morrisonicola : Effects of operational parameters, kinetics and characterization

Abstract Pinus morrisonicola has been reported bio-benefit and contains high amount of antioxidant bioactive compounds. This study followed a previous work to pretreat the Pinus morrisonicola by compression-puffing process and then extracted the antioxidant phenolic compounds from the pretreated samples for the measurements. The effects of ethanol concentration, extraction temperature and liquid–solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (DPPH scavenging activity, FRAP and ABTS scavenging activity) are reported for demonstration the pretreatment efficiencies. Semi-qualitative characterization of the extract compounds was conducted.

Antioxidant phenolic compounds from Pinus morrisconicola using compressional-puffing pretreatment and water–ethanol extraction: Optimization of extraction parameters

This study is aimed to increase the extraction yield and efficiency of the antioxidant phenolic compounds from Pinus morrisonicola. The optimum extraction conditions for the phenolic compounds from pine needles are determined by using response surface methodology with a central-composite design. Effects of ethanol concentration, extraction temperature and liquid–solid ratio on the extraction yield (total flavonoids content and total phenolic content) and antioxidant activity (2,2′-diphenyl-1-picryhydrazyl (DPPH) scavenging activity, ferric reducing antioxidant power (FRAP) and 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) scavenging activity) are investigated. The optimum conditions to maximize the extraction yields and antioxidant activity of the pine needles extracts are: 36.1% v/v of ethanol, 70ºC and 50ml/g of liquid–solid ratio. The pine needles extracts are shown to be free of 311 tested pesticide residues or certain heavy metals and exhibit comparable quantities of total flavonoids contents, total phenolic contents and the antioxidant activities with commercial products.

Antioxidant Properties of Essential Oil Extracted from Pinus morrisonicola Hay Needles by Supercritical Fluid and Identification of Possible Active Compounds by GC/MS.

Pine (Pinus morrisonicola Hay, PM) needles have been used as folk medicine for their antihypertension and lipid-lowering effects. As supercritical fluid extraction (SFE) is considered an ideal technique for the extraction of essential oil from plant materials, the present work investigated the optimal SFE conditions and the protective effects of different resulting fractions of PM needles on lipid peroxidation and foam cell production in macrophages. Nine PM needle extracts (PME1–9) were obtained in 1%–4% yields using different SFE conditions, of which PME1 had the lowest yield (1.1%) and PME3 the highest (3.9%). PME3 exhibited lower cytotoxic effects and stronger inhibition of lipid peroxidation and formation of foam cell in RAW 264.7 macrophages than those of other PME extracts. PME3-1 purified from PME3 by column and thin layer chromatography inhibited LDL oxidation more effectively than did PME3 in a cell-free system oxidized by Cu2+. PME3-1 dose-dependently (25–100 μg/mL) decreased conjugated diene levels and foam cell formation induced by ox-LDL. GC/MS analyses revealed that 1-docosene, neophytadiene, and methyl abietate were increased 5.2-, 1.7- and 4.3-fold in PME3-1 relative to PME3. A new hydrocarbon compound, cedrane-8,13-diol, was identified in PME3-1. Overall, the present study demonstrates the optimal extraction conditions of SFE of PM and identifies the most potent antioxidant fractions and possible active compounds in PM.

Pine (Pinus morrisonicola Hayata) needle extracts sensitize GBM8901 human glioblastoma cells to temozolomide by downregulating autophagy and O(6)-methylguanine-DNA methyltransferase expression.

Pine needle extracts of Pinus morrisonicola (Hayata) are commonly used as a functional health beverage. However, it remains unclear what the mechanism is underlying the antitumor activity of pine needle extract. The aims of present study were to investigate the anti-glioblastoma effects of pine needle extracts as well as its bioactive compounds. From three different solvent extracts of pine needles, the water extract displayed the strongest cytotoxicity effects on GBM8901 glioblastoma cells. The isolated compounds were identified as pinocembrin, chrysin, and tiliroside. Chrysin was the most active ingredient of pine needle extract for the induction of apoptosis and suppression of migration and invasion. It also markedly inhibited temozolomide (TMZ)-induced autophagy and O(6)-methylguanine-DNA methyltransferase (MGMT) expression. Because both autophagy and MGMT overexpression have been implicated to TMZ-induced drug resistance in glioblastoma, our results showed that pine needle extract and chrysin may serve as a potential anticancer agent against glioblastoma, especially with regard to sensitizing glioblastoma cells resistant to TMZ.

192: Pinus morrisonicola Hayata extracts inhibit cell proliferation and promote apoptosis of human promyelocytic HL-60 leukemia cells

192: Pinus morrisonicola Hayata extracts inhibit cell proliferation and promote apoptosis of human promyelocytic HL-60 leukemia cells

Carboxylic acids in PM2.5 over Pinus morrisonicola forest and related photoreaction mechanisms identified via Raman spectroscopy

Abstract The PM 2.5 aerosol from within an area of Pinus morrisonicola Hayata in Taiwan was collected and analyzed for its low molecular weight carboxylic acid (LMWCAs) content. Oxalic acid was the major LMWCA in the aerosol, followed by acetic, tartaric and maleic acids. This differs significantly from the LMWCA composition of PM 2.5 aerosol reported for a southern Taiwan suburban region (oxalic > succinic > malonic) [Atmospheric Environment 42, 6836–6850 (2008)]. P. morrisonicola Hayata emits oxalic, malic and formic acids and yet there was an abundance of maleic and tartaric acids in the PM 2.5 forest aerosol, indicating that tartaric acid is derived from the transformation of other P. morrisonicola Hayata emissions. Raman spectroscopy was applied and 28 species of LMWCAs and inorganic species were identified. The photochemical mechanisms of maleic and tartaric acids were studied and it was found that the abundant tartaric acid in forest aerosol is most probably the photochemical product from reactions of maleic acid. Furthermore, tartaric acid is photochemically transformed into formic acid and ultimately into CO 2 .

Antioxidant properties and mutagenicity of Pinus morrisonicola and its vinegar preparation

Pine ( Pinus morrisonicola Hayata) needles was tested for toxicity and mutagenicity by Ames test and reducing power and scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals. Based on the tests on Salmonella typhimurium TA97 and TA100, P. morrisonicola extract was not toxic and mutagenic. The reducing powers of P. morrisonicola extract and ascorbic acid were comparable with both effective concentration (EC 50) values being 0.06 mg/mL. The extract was more effective in scavenging ability and EC 50 values were 0.02 and 0.04 mg/mL for P. morrisonicola extract and ascorbic acid, respectively. With P. morrisonicola powder (50 mg/mL) added to the cultured saccharified broth at day 1, the wine with an ethanol content of 8.23% was produced at day 12. After 11 days of fermentation, the vinegar was obtained with an acidity of 5.21 g/100 mL. Accordingly, P. morrisonicola could be used in the development of alcoholic and vinegar products to provide its beneficial health effects.

Enzymatic hot pressurized fluids extraction of polyphenolics from Pinus taiwanensis and Pinus morrisonicola

This study was engaged in application of enzymatic hydrolysis on the hot pressurized fluid extraction (HPFE) to isolate the polyphenols from the needles of Pinus taiwanensis (PT) and Pinus morrisonicola (PM). This HPFE process was investigated using an L 18 orthogonal experimental design. Our experimental results indicate that a mixed enzyme of cellulase, hemicellulase, pectinase and protease is able to enhance cell breakage during the sample pretreatment and the HPFE process. A suitable dynamic enzymatic HPFE conditions were 1.5% wt of mixed enzymes in 70% ethanol (pH 6) at a flow rate of 10 mL/min, under pressure 689 kPa and 343 K for 180 min. The content of polyphenols obtained from the dynamic mixed enzymatic hydrolysis HPFE process is higher than that obtained from the static enzymatic hydrolysis HPFE process at 4137 kPa and 343 K for 180 min. Anti-oxidative activities of the ethyl acetate fraction from partition of the resultant of the dynamic enzymatic HPFE process determined by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, peroxide radical release inhibition, nitrite oxide radical scavenging and photochemiluminescence were highly correlated with the content of polyphenols in the extracts. The extracts of PT needles have higher chemical antioxidant activities than that obtained from PM needles . However, in terms of antioxidant abilities from water-soluble and lipid-soluble fractions determined by photochemiluminescence method, the PM needles extracts demonstrated better activity than that from the PT needles extract. In the bioassay tests, the extracts of PM and PT needles demonstrate only a little viability of normal control cells at doses of 8.59 μg/mL and 5.86 μg/mL, respectively, but exhibit 10% and 16% of growth inhibition effect on DBTRG-glioblastoma cell of the human brain.

Protective effect of pine ( Pinus morrisonicola Hay.) needle on LDL oxidation and its anti-inflammatory action by modulation of iNOS and COX-2 expression in LPS-stimulated RAW 264.7 macrophages

The protective effects of pine ( Pinus morrisonicola Hay.) needle on low-density lipoprotein (LDL) oxidation and nitric oxide production in macrophages as well as its bioactive compounds were investigated. Of the four solvent extracts, the ethyl acetate extract of pine needle (EAE-PN) exhibited the strongest scavenging action on free radicals. EAE-PN significantly inhibited copper-induced LDL oxidation through prolonging the lag phase of conjugated dienes formation and decreasing the relative electrophoretic mobility of LDL. Lipid accumulation and foam cell formation were significantly reduced when EAE-PN (75 μg/mL) was added to the medium co-incubated with macrophages cells and copper-induced LDL. EAE-PN also markedly inhibited reactive oxygen species production in RAW 264.7 cells stimulated with lipopolysaccharide (LPS). As regards NO production in cells, EAE-PN showed dose-dependent inhibitory effect on nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. The inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein expressions in LPS-stimulated RAW 264.7 cells were inhibited by EAE-PN. RT-PCR analysis indicated that the iNOS and COX-2 mRNA expression were suppressed by EAE-PN. The major phenolic compounds in EAE-PN were epicatechin and p-coumaric acid by HPLC analysis. The presence of epicatechin and p-coumaric acid in EAE-PN may be partially responsible for the biological action of EAE-PN. Taken together, these results suggest that EAE-PN may provide potential protective effects against LDL oxidation and attenuating excessive NO generation at inflammatory sites; consequently, this may contribute to anti-atherosclerotic and anti-inflammatory effects of EAE-PN.