• Natural Product Sciences
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• v.16 no.2
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• pp.123-132
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• 2010

The aim of the present study was to investigate whether self-fermented pine extract for 2 years (SFPE2) and ethyl acetate (EtOAc) fraction from self-fermented pine needle extract may affect the contractility of the isolated aortic strips and blood pressure of normotensive rats. SFPE2 ($360-1440\;{\mu}g/mL$) significantly depressed both phenylephrine ($10\;{\mu}M$)- and high potassium (56 mM)-induced contractile responses of the isolated rat aortic strips in dose-dependent fashion. The EtOAc-fraction ($400\;{\mu}g/mL$) also inhibited both phenylephrine ($10\;{\mu}M$)- and high potassium (56 mM)-induced contractile responses. Also, in anesthetized normotensive rats, intravenous injection of the EtOAc fraction (1.0~10.0 mg/kg) dose-dependently elicited hypotensive responses. The EtOAc fractions (1.0 and 3.0 mg/kg/30 min) inhibited norepinehrine-induced pressor responses. Intravenous infusion of SFPE2 fraction (3.0 and 10.0 mg/kg/30 min) also inhibited norepinehrine-induced pressor responses in both anesthetized spontaneously hypertensive rats (SHRs) and normotensive rats. In conclusion, these results suggest that both SFPE2 and the EtOAc fraction cause vascular relaxation in the aortic strips isolated from normotensive rats and SHRs as well as vasodepressor responses. Based on these experimental data, it seems that SFPE2 or the EtOAc fraction possesses active antihypertensive components, which are available to prevent or treat hypertension in future.

• Natural Product Sciences
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• v.19 no.4
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• pp.290-296
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• 2013

In preliminary tests, we examined the effect of several fractions isolated from fermented pine needle extract on pacemaker potentials in cultured interstitial cells of Cajal (ICCs) from the mouse colon using a whole cell patch clamp technique. Among these fractions, Fraction 3 (F3) elicited the most powerful depolarization of membrane. Therefore, the aim of the present study was to investigate the effect of F3 obtained from fermented extract of Pinus densiflora needle on pacemaker potentials in ICCs and to establish its mechanism of action. Colonic ICCs generated spontaneous periodic pacemaker potentials in the current-clamp mode. F3 depolarized the membrane and decreased the frequency and amplitude of pacemaker potentials in a dose-dependent fashion. The F3-induced effects on pacemaker potentials were blocked by methoctramine, a muscarinic $M_2$ receptor antagonist, and by glycopyrrolate, a muscarinic $M_3$ receptor antagonist. The F3-induced effects on pacemaker potentials were blocked by external $Na^+$-free solution and by flufenamic acid, a non-selective cation channel blocker, as well as by the removal of external $Ca^{2+}$ and in the presence of thapsigargin, a $Ca^{2+}$-ATPase inhibitor in the endoplasmic reticulum. Taken together, these results suggest that F3 of pine needle extract modulates the pacemaker activity of colonic ICCs by the activation of non-selective cation channels via muscarinic $M_2$ and $M_3$ receptors. And external $Ca^{2+}$ influx and intracellular $Ca^{2+}$ release are involved in F3 actions on ICCs.

• Korean Journal of Food Science and Technology
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• v.27 no.6
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• pp.978-984
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• 1995

This study was conducted to investigate the physiological functionality of the hot water and 70% acetone extracts obtained from the pine needle and mugwort. Flavanol tannin content was above 60% of the total polyphenol in pine needle extracts. 70% Acetone extract from pine needle exhibited inhibition percentage of about 82.2% on the mutagenicity of Trp-P-1. Angiotensin I converting enzyme(ACE) inhibition activity was 61% and 50% in the hot water extract and in the 70% acetone extract respectively. Electron donating ability(EDA) of the hot water and 70% acetone extracts obtained from the pine needle was significantly good above 80%. 70% Acetone extract from pine needle showed inhibitary effect against the polyphenol oxidase(PPO) of water dropwort. The nitrite scavenging ability was appeared in all the extracts examined and it showed $80{\sim}90%$ at pH 3.0. The oxidative stability was determined by POV. Results showed that ethyl acetate fraction is better antioxidants than chloroform and butanol fractions in the soybean oil. Then, in the mugwort extracts, antimutagenicity of 70% acetone extract was similar to that of pine needle. Ratio of ACE inhibition activity was higher than that of pine needle extracts. EDA of hot water and 70% acetone extracts was 45%. Extracts from the mugwort were lower in nitrite scavenging ability about $20{\sim}30%$ than extracts from pine needle.

• Preventive Nutrition and Food Science
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• v.15 no.1
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• pp.36-43
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• 2010

The purpose of this study was to investigate the phenolic acid, proanthocyanidin (PAs), and flavonol glycoside contents, as well as the antioxidant activities of pine needle extracts from six species of young pine trees. The extracts were prepared from Section Pinus (Diploxylon): P. densiflora, P. sylvestris, P. pinaster and P. pinea, and Section Strobus (Haploxylon): P. koraiensis and P. strobus. Phenolics were extracted from pine needles with 80% acetone to obtain the soluble free fraction, and insoluble residues were digested with 4 M NaOH to obtain bound ethyl acetate and bound water fractions. Phenolics were analyzed by HPLC, and the hydrophilic antioxidant activity was measured using oxygen radical absorbance capacity (ORAC). Total phenolic and flavonoid contents of the soluble free fraction were higher than those of the bound ethyl acetate and bound water fractions. The main phenolics were monomers and polymers of PAs in the soluble free fraction, and phenolic acids and flavonol glycosides in bound ethyl acetate fraction. Flavonol glycosides found in different species of pine needles were qualitatively similar within fractions, but composition varied among Pinus sections. High levels of kaempferol arabinoside and an unknown compound were present in all Strobus species. The soluble free fraction had the highest antioxidant activity, followed by bound ethyl acetate and bound water fractions.

• Journal of Microbiology and Biotechnology
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• v.31 no.1
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• pp.137-143
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• 2021

Most cervical cancers are associated with high-risk human papillomavirus (HPV) infection. Currently, cervical cancer treatment entails surgical removal of the lesion, but treatment of infection and preventing tissue damage are issues that still remain to be addressed. Herbal medicine and biological studies have focused on developing antiviral drugs from natural sources. In this study, we analyzed the potential antiviral effects of Pinus densiflora Sieb. et Zucc. leaf extracts against HPV. The pine needle extracts from each organic solvent were analyzed for antiviral activity. The methylene chloride fraction (PN-MC) showed the highest activity against HPV pseudovirus (PV). The PN-MC extract was more effective before, rather than after treatment, and therefore represents a prophylactic intervention. Mice were pre-treated with PN-MC via genital application or oral administration, followed by a genital or subcutaneous challenge with HPV PV, respectively. The HPV challenge results showed that mice treated via genital application exhibited complete protection against HPV. In conclusion, PN-MC represents a potential topical virucide for HPV infection.

• Natural Product Sciences
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• v.19 no.1
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• pp.36-48
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• 2013

The aim of the present study was to investigate the effects of several fractions obtained from methylene chloride ($CH_2Cl_2$) extract of self-fermented pine needle (SFPNE) on the acetylcholine (ACh)-evoked CA release from the isolated perfused model of the rat adrenal medulla and to establish the mechanism of the most active fraction (Fr.)-induced inhibitory action on the CA release. We obtained 6 fractions from $CH_2Cl_2$ extract of self-fermented pine needle. For the ACh (5.32 mM)-evoked CA release, the following rank order of inhibitory potency was obtained: Fr.4-5 > Fr.8-11 ${\gg}$ Fr.3 > Fr.6 = Fr.7 > Fr.1-2. Fr. 4 - 5 (60 ${\mu}g/mL$) perfused into an adrenal vein for 90 min produced relatively time-dependent inhibition of the CA secretory responses to ACh (5.32 mM), DMPP (100 ${\mu}M$), McN-A-343 (100 ${\mu}M$) and high $K^+$ (56 mM). Fr. 4 - 5 itself did not affect basal CA secretion. Also, in the presence of Fr. 4 - 5 (60 ${\mu}g/mL$), the CA secretory responses to angiotensin II (AngII, 0.1 ${\mu}M$), veratridine (50 ${\mu}M$), Bay-K-8644 (10 ${\mu}M$), and cyclopiazonic acid (10 ${\mu}M$) were significantly reduced, respectively. In the simultaneous presence of Fr. 4 - 5 (60 ${\mu}g/mL$) and L-NAME (30 ${\mu}M$), the inhibitory responses of Fr. 4 - 5 on the CA secretion evoked by ACh, DMPP, high $K^+$, AngII, Bay-K-8644 and veratridine were considerably recovered to the extent of the corresponding control secretion compared with that of Fr. 4 - 5-treatment alone. The level of NO released from adrenal medulla after the treatment of Fr. 4 - 5 (60 ${\mu}g/mL$) was greatly elevated compared with the basal level. Taken together, these results demonstrate that Fr. 4 - 5 inhibits the CA secretion from the isolated perfused rat adrenal medulla evoked by stimulation of cholinergic receptors as well as by direct membrane-depolarization. It seems that this inhibitory effect of Fr. 4 - 5 is mediated by blocking the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells as well as by inhibition of $Ca^{2+}$ release from the cytoplasmic calcium store, which is evoked at least partly through the increased NO production due to the activation of NO synthase. Based on these results, it is also thought that Fr. 4 - 5 isolated from $CH_2Cl_2$ extract of pine needle may contain beneficial antihypertensive components to prevent or treat hypertension.

• Journal of Life Science
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• v.13 no.5
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• pp.684-691
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• 2003

This study was designed to investigate the effects of ethyl acetate (EtOAc) fraction of pine (Pinus densiflora Sieb et Zucc) needle extract on membrane fluidity (MF), basal and induced oxygen radical (BOR and IOR), lipid peroxide(LPO) and oxidized protein (OP) as an oxidative stress, and lipofuscin(LF) in liver membranes of male Sprague-Dawley rats. Rats were fed basic diets (control) and experimental diets (EtOAc-25, EtOAc-50 and EtOAc-100) for 45days. MFs were significantly increased (about 10%) in mitochondria of EtOAc-100 group compared with control group. BOR and IOR formations in mitochondria were significantly inhibited (about 12∼18% and 9 ∼l2%, respectively) in EtOAc-50 and EtOAc-100 groups, while BOR and IOR formations in microsomes were significantly inhibited (about 9∼l3% and 18∼19%, respectively) compared with control group. LPO levels were significantly inhibited (about 10% and 12∼13%, respectively) in mitochondria of EtOAc-100 and microsomes of EtOAc-50 and EtOAc-100 groups, whereas OP levels were significantly inhibited (about 13∼14%) in mitochondria of EtOAc-50 and EtOAc.-100 groups compared with control group. LF formations were significantly inhibited (about 10∼14%) in these three EtOAc groups. These results suggest that ethyl acetate fraction of pine needle may play an effective role in attenuating an oxidative stress and increasing a membrane fluidity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.7
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• pp.1082-1087
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• 2003

This study was designed to investigate the effects of butanol (BuOH) fraction of pine (Pinus densiflora Sieb et Zucc) needle extract on membrane fluidity (MF), basal and induced oxygen radicals (BOR and IOR), lipid peroxide (LPO) and oxidized protein (OP) as an oxidative stress, and lipofuscin (LF) in liver membranes of Sprague-Dawley male rats. The rats were fed basic diets (control group) and experimental diets (BuOH-25, BuOH-50 and BuOH-100) prepared with 25, 50 and 100 mg added to basic diet for 45 days. MFs were significantly increased (about 16∼22%) in mitochondria of BuOH-50 and BuOH-100 groups compared with control group (p<0.01∼0.001) BOR and IOR formations in mitochondria were significantly decreased (11∼17% and 11∼28%, respectively) in these three BuOH groups (p<0.05∼0.001), while BOR and IOR formations in microsomes were significantly decreased (11∼24%) in BuOH-50 and BuOH-100 groups, and (15∼24%) in these three BuOH groups compared with control group (p<0.05∼0.001; p<0.01-0.001). LPO levels were significantly decreased (9% and 9∼13%, respectively) in mitochondria of BuOH-100 and microsomes of BuOH-50 and BuOH-100 groups (p<0.05∼0.01), whereas OP levels were significantly decreased (10∼12%) in mitochondria of BuOH-50 and BuOH-100 groups compared with control group (p<0.05). LF formations were significantly decreased (8∼9%) in BuOH-50 and BuOH-100 groups (p<0.05). These results suggest that butanol fraction of pine needle extract may playa effective role in an attenuating an oxidative stress and increasing a membrane fluidity.

• Korean Journal of Food Science and Technology
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• v.30 no.2
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• pp.450-455
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• 1998

Pine has been known as a traditional medicinal plant and as showing a physically beneficial function to a human being. Therefore, this study was performed to investigate the physiological activities of main pine neddles. Ethanol extracts from pinus needles did net exhibit any mutagenicity. On the contrary, inhibitory effects of ethanol extract were observed on mutagenicity induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide (4NQO), 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1) and benzo(a)pyrene $(B({\alpha})P)$ using Salmonella typhimurium reversion assay. On direct-acting mutagen (MNNG, 4NQO) and indirect-acting mutagen (Trp-P-1, $(B({\alpha})P)$, we observed higher inhibitory effect. Stepwise fractionation of the ethanol extract was done by using ether, chloroform, ethyl acetate, butanol and water to obtain effective fraction. Among them, water fractions $(100\;{\mu}g/plate)$ of Pinus thunbergii, Pinus rigida, Pinus densiflora and Pinus koraiensis showed high inhibition of 91.65%, 94.7%, 84.22% and 79.02%, respectively, on the mutagenicity of MNNG in Salmonella typhimurium TA100.

• Journal of Nutrition and Health
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• v.35 no.3
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• pp.296-302
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• 2002

This study was designed to investigate the effects of a butanol extract of pine needles (Pinus densiflora Sieb et Zucc) on lipid metabolism and oxidative stress in rats. Twenty-eight male Sprague-Dawley (SD) rats were divided into four groups over a 45 days study period: the control group on a basic diet, and three experimental groups on three different dietary levels of the butanol fraction, specifically 25 mg (BuOH-25), 50 mg (BuOH-50), and 100 mg (BuOH-100) butanol fraction/kg body weight/day, thereby 0.025%, 0.05%, 0.1% of butanol extract of pine needles was added to basic diet respectively. At the end of the experimental period, body weights and food intakes were food intakes were not different among the four groups. Total and LDL-cholesterol levels were markedly decreased in the BuOH-25, BuOH-50, and BuOH-100 groups, respectively, as follows: 12.8%, 19.1% and 21.6% reductions in total cholesterol; and 10.2%, 15.6% and 23.7% reductions in LDL-cholesterol. However, HDL-cholesterol levels were significantly increased (by approximately 20%) in the serum of the BuOH-100 group only, compared with the control and other experimental groups. Atherogenic indices were also markedly decreased in the three experimental groups, by 24.8%, 30.4% and 36.2%, for each of the BuOH-25, BuOH-50, and BuOH-100 groups, respectively, compared with the control group. The levels of the hydroxyl radical (·OH) and of lipid peroxide (LPO) in the serum of the three experimental groups were significantly reduced, by 9.8%, 19.7% and 21.2%; and by 13.3%, 13.3% and 16.7%, for the BuOH-25, BuOH-50, and BuOH-100 groups, respectively Significant increases in serum superoxide dismutase (SOD) were observed in the BuOH-50 and BuOH-100 groups; specifically, 12.1% in the BuOH-50 group and 23.3% in the BuOH-100 group, compared with the control group. Significant increases in catalase (CAT) avtivities, of 24.7% in the BuOH-50 group and 29.2% in the BuOH-100 group, were also observed, compared to the control group. These results suggest that a butanol extract of pine needles could inhibit chronic degenerative disease through improving lipid metabolism, and could also effectively modulate the aging process attenuating oxidative stress.