• Journal of Life Science
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• v.32 no.5
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• pp.348-354
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• 2022

The leaves, stems, seeds, and roots of Dendropanax morbifera have been used since ancient times as folk medicines for the treatment of headaches, skin diseases, infectious diseases, and other ailments. This study investigated the antioxidant, alcohol metabolism, and hepatoprotective effects of D. morbifera leaf and stem water extracts. The total polyphenol content of the D. morbifera leaf and stem water extracts was 49.56 mg tannic acid equivalent (TAE)/g, and the 1,1-diphenyl-2-picrylhydrazy (DPPH) radical scavenging activity of the D. morbifera leaf and stem water extracts was 84.09% at 1,000 ㎍/ml concentration. The effects of D. morbifera leaf and stem water extracts on alcohol metabolism were determined by measuring the generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). The ADH and ALDH activities of D. morbifera leaf and stem water extracts were increased in a dose-dependent manner at 37.68% and 41.67%, respectively, at a 1,000 ㎍/ml concentration. The D. morbifera leaf and stem water extracts showed significant protective effects against tacrine-induced cytotoxicity in HepG2 cells at 50 ㎍/ml. Based on our results, we concluded that D. morbifera leaf and stem water extracts may be used as major pharmacological agents, such as antioxidants, alcohol metabolism, and anti-hepatitis remedies.

• Biomolecules & Therapeutics
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• v.31 no.1
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• pp.73-81
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• 2023

Sirtuins (SIRTs) belong to the nicotinamide adenine dinucleotide (NAD+)-dependent class III histone deacetylase family. They are key regulators of cellular and physiological processes, such as cell survival, senescence, differentiation, DNA damage and stress response, cellular metabolism, and aging. SIRTs also influence carcinogenesis, making them potential targets for anticancer therapeutic strategies. In this study, we investigated the anticancer properties and underlying molecular mechanisms of a novel SIRT1 inhibitor, MHY2251, in human colorectal cancer (CRC) cells. MHY2251 reduced the viability of various human CRC cell lines, especially those with wild-type TP53. MHY2251 inhibited SIRT1 activity and SIRT1/2 protein expression, while promoting p53 acetylation, which is a target of SIRT1 in HCT116 cells. MHY2251 treatment triggered apoptosis in HCT116 cells. It increased the percentage of late apoptotic cells and the sub-G1 fraction (as detected by flow cytometric analysis) and induced DNA fragmentation. In addition, MHY2251 upregulated the expression of FasL and Fas, altered the ratio of Bax/Bcl-2, downregulated the levels of pro-caspase-8, -9, and -3 proteins, and induced subsequent poly(ADP-ribose) polymerase cleavage. The induction of apoptosis by MHY2251 was related to the activation of the caspase cascade, which was significantly attenuated by pre-treatment with Z-VAD-FMK, a pan-caspase inhibitor. Furthermore, MHY2251 stimulated the phosphorylation of c-Jun N-terminal kinase (JNK), and MHY2251-triggered apoptosis was blocked by pre-treatment with SP600125, a JNK inhibitor. This finding indicated the specific involvement of JNK in MHY2251-induced apoptosis. MHY2251 shows considerable potential as a therapeutic agent for targeting human CRC via the inhibition of SIRT1 and activation of JNK/p53 pathway.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.199-209
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• 2023

Background: Due to the interrupted blood supply in cerebral ischemic stroke (CIS), ischemic and hypoxia results in neuronal depolarization, insufficient NAD+, excessive levels of ROS, mitochondrial damages, and energy metabolism disorders, which triggers the ischemic cascades. Currently, improvement of mitochondrial functions and energy metabolism is as a vital therapeutic target and clinical strategy. Hence, it is greatly crucial to look for neuroprotective natural agents with mitochondria protection actions and explore the mediated targets for treating CIS. In the previous study, notoginseng leaf triterpenes (PNGL) from Panax notoginseng stems and leaves was demonstrated to have neuroprotective effects against cerebral ischemia/reperfusion injury. However, the potential mechanisms have been not completely elaborate. Methods: The model of middle cerebral artery occlusion and reperfusion (MCAO/R) was adopted to verify the neuroprotective effects and potential pharmacology mechanisms of PNGL in vivo. Antioxidant markers were evaluated by kit detection. Mitochondrial function was evaluated by ATP content measurement, ATPase, NAD and NADH kits. And the transmission electron microscopy (TEM) and pathological staining (H&E and Nissl) were used to detect cerebral morphological changes and mitochondrial structural damages. Western blotting, ELISA and immunofluorescence assay were utilized to explore the mitochondrial protection effects and its related mechanisms in vivo. Results: In vivo, treatment with PNGL markedly reduced excessive oxidative stress, inhibited mitochondrial injury, alleviated energy metabolism dysfunction, decreased neuronal loss and apoptosis, and thus notedly raised neuronal survival under ischemia and hypoxia. Meanwhile, PNGL significantly increased the expression of nicotinamide phosphoribosyltransferase (NAMPT) in the ischemic regions, and regulated its related downstream SIRT1/2/3-MnSOD/PGC-1α pathways. Conclusion: The study finds that the mitochondrial protective effects of PNGL are associated with the NAMPT-SIRT1/2/3-MnSOD/PGC-1α signal pathways. PNGL, as a novel candidate drug, has great application prospects for preventing and treating ischemic stroke.

• Journal of Nutrition and Health
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• v.57 no.4
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• pp.403-417
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• 2024

Purpose: This study investigated the anti-inflammatory effects of Evodiae Fructus (EF) on hydrochloric acid (HCl)/ethanol-induced gastritis, focusing on its impact on oxidative stress by analyzing inflammatory cytokines and inflammation-related factors. The total polyphenol and flavonoid contents were determined through in vitro experiments, while the radical scavenging activity was confirmed using 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) assays. Methods: In vivo experiments were conducted on rats divided into 5 groups (n = 7/in each group): normal group (Normal), 150 mM HCl/60% ethanol-induced gastritis group (Control), 150 mM HCl/60% ethanol-induced gastritis group administered 10 mg/kg sucralfate (SC), 150 mM HCl/60% ethanol-induced gastritis group administered EF at the doses of 100 mg/kg or 200 mg/kg (EF100 or EF200). The mice were pretreated with the extract (EF) or drug (SC), and after 1 hour, 150 mM HCl/60% ethanol (v/v) mixture was administered orally. Reactive oxygen species (ROS) levels, peroxynitrite (ONOO^-), and pro-inflammatory cytokines including tumor necrosis factor-α and interleukin-1 beta were assessed in serum. Additionally, western blotting of the gastric tissues confirmed the expression of inflammation-related proteins. Results: EF alleviated the gastric mucosal damage caused by 150 mM HCl/60% ethanol. The assessment of oxidative stress in the serum showed that EF significantly reduced ROS and ONOO^- levels and significantly decreased the levels of pro-inflammatory cytokines. Western blot analysis revealed that EF reduced ROS-generating nicotinamide adenine dinucleotide phosphate oxidase subunits, including gp91^phox, p22^phox, and p47^phox. Additionally, EF mitigated the inflammation by inhibiting the mitogen-activated protein kinase signaling pathway. Conclusion: These results indicate that EF is a potential herbal medicine candidate for the treatment of oxidative stress-induced gastritis.

• Journal of Acupuncture Research
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• v.20 no.5
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• pp.118-132
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• 2003

Objective: The aim of this study was to investigate the effects of Joksamni(ST36) combination on NAD PH-diaphorase, neuronal nitric oxide synthase(nNOS), neuropeptide Y(NPY) and vasoactive intestinal peptide (VIP) in the cerebral cortex of spontaneously hypertensive rat. Methods: The experimental groups were divided into four groups: Normal, Joksamni(ST36), Joksamni(ST36)+Eumneungcheon(SP9), and Joksamni(ST36)+Gokji(LI11). Needles were inserted into acupoints at the depth of 0.5cm with basic insertion method. Electroacupuncture was done under the condition of 2Hz electrical biphasic pulses with continuous rectangular wave lasting for 0.2ms until the muscles produced visible contractions. Such stimulation was applied continuously for 10 minutes, 1 time every 2 days for 10 sessions of treatments. Thereafter we evaluated changes in NADPH-d positive neurons histochemically and changes in nNOS, NPY and VIP positive neurons immunohistochemically. Results: The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in primary somatosensory cortex, visual cortex, auditory cortex, perirhinal cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of NADPH-d positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group with the exception of primary somatosensory cortex. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group in all areas of cerebral cortex and Joksamni(ST36)+Gokji(LI11) group in auditory cortex, perirhinal cortex, insular cortex were significantly increased as compared to the Joksamni(ST36) group. The optical densities of nNOS positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. The optical densities of NPY positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in primary motor cortex, primary somatosensory cortex, cingulate cortex as compared to the Joksamni (ST36) and Joksamni(ST36)+Eumneungcheon(SP9) groups. The optical densities of VIP positive neurons of the Joksamni(ST36)+Eumneungcheon(SP9) group were significantly increased in all areas of cerebral cortex except for cingulate cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in auditory cortex, cingulate cortex, perirhinal cortex as compared to the Joksamni(ST36) group. The optical densities of VIP positive neurons of the Joksamni(ST36)+Gokji(LI11) group were significantly decreased in all areas of cerebral cortex as compared to the Joksamni(ST36)+Eumneungcheon(SP9) group. Conclusions: The result demonstrated that electroacupuncture on Joksamni(ST36) and its combination change the activities of the NO system and peptidergic system in the cerebral cortex of SHR and that acupoint combination is one of the important parameters for the effects.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.5
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• pp.655-661
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• 2010

The nutraceutical role of fermented omija (Schizandra chinensis) beverage (FOB) was determined through the analysis of fibrinolytic and alcohol metabolizing activities, nitrite scavenging activity, and angiotensin converting enzyme and $\alpha$-glucosidase inhibitory effects. Firstly, FOB increased fibrinolytic activity in a dose-dependent manner and indicated angiotensin converting enzyme inhibitory activity of 94.8% at 20% FOB (0.6 mg/mL). In addition, the inhibitory activities of FOB on $\alpha$-amylase and $\alpha$-glucosidase were determined to be 100% at 100% FOB (3 mg/mL) and 49% at 60% FOB (1.8 mg/mL), respectively. Nitrite scavenging activity of FOB was about 96.1%, 72.3%, and 68.3% on pH 1.2, 3.0, and 6.0 at 100% FOB, respectively. To determine influence of FOB on alcohol metabolism, the generating activities of reduced-nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) were measured. Facilitating rate of ADH activity was 70.3% at 50% FOB, but ALDH activity was not affected. These results revealed that FOB has a strong alcohol metabolizing activity, and fibrinolytic and nitrite scavenging activities and exhibits the angiotensin converting enzyme, $\alpha$-amylase, and $\alpha$-glucosidase inhibitory activities.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1096-1101
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• 2008

Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.6
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• pp.819-827
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• 2016

This study was carried out to investigate anti-inflammatory, anti-diabetic, alcohol metabolizing, and hepatoprotective effects of hot water (MOW) and 80% ethanol (MOE) extracts from moringa (Moringa oleifera Lam.) leaf. The total phenol content of MOW and MOE were 45.49 and 63.06 mg tannic acid equivalents/g, respectively. 1,1-Diphenyl-2-picrylhydrazyl radical scavenging activities of MOW and MOE were remarkably elevated in a dose-dependent manner, and about 60.8% and 71.3% at 1 mg/mL, respectively (P<0.01). Superoxide dismutase-like activities of MOW and MOE were 2.8% and 7.4% at 5 mg/mL, respectively (P<0.05). ${\alpha}-Glucosidase$ inhibitory activity also increased in a dose-dependent manner in both extracts, and MOE was higher about two times than MOW at 5 mg/mL (P<0.001). The effects of MOW and MOE on alcohol metabolizing activity were determined by measuring generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). ADH and ALDH activities significantly increased upon addition of MOW and MOE (P<0.05). Anti-inflammatory activity was examined in lipopolysaccharide-stimulated RAW 264.7 cells. Nitric oxide production was reduced to 32.1% and 81.2% by addition of MOW and MOE at 1 mg/mL, respectively (P<0.05). MOW and MOE showed significant protective effects against tacrine-induced cytotoxicity in Hep3B cells at $100{\mu}g/mL$. These results suggest that moringa leaf extracts have great potential as natural health products.

• Analytical Science and Technology
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• v.23 no.6
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• pp.531-536
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• 2010

A quantitative analytical method has been established for the measurement of inosine 5'-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a ${\beta}$-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5'-monophosphate (IMP) into xanthosine 5'-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and $NAD^+$ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear ($r^2$=0.999999) in the range of $0.2-50.0\;{\mu}M$ and the limit of quantification (LOQ) was $0.2\;{\mu}M$. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%, respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = $27.70{\pm}6.28\;nmol/h/mg$ protein).

• The Korean Journal of Nuclear Medicine
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• v.36 no.6
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• pp.333-343
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• 2002

Purpose: A new linker, hydrazino nicotinamide (HYNIC), was recently introduced for labelling of liposome with $^{99m}Tc$. In this study we synthesized HYNIC derivatized PEG (polyethylene glycol)-liposomes radiolabeled with $^{99m}Tc$. Materials and Methods: In order to synthesize HYNIC-DSPE (distearoyl phosphatidyl ethanolamine) which is a crucial component for $^{99m}Tc$ chelation, first of all succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized from 6-chloronicotinic acid by three sequential reactions. A DSPE derivative of succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was transformed into HYNIC-DSPE by HCI/dioxane. HYNIC-PEG-liposomes were prepared by hydration of the dried lipid mixture of EPC (egg phosphatidyl choline): PEG-DSPE : HYNIC-DSPE:cholesterol (1.85:0.15:0.07:1, molar ratio). The HYNIC-PEG-liposomes were labeled with $^{99m}Tc$ in the presence of $SnCl_2{\cdot}2H_2O$ (a reducing agent) and tricine (a coligand). To investigate the level of in vivo transchelation of $^{99m}Tc$ in the liposomes, the $^{99m}Tc$-HYNiC-PES-liposomes were incubated with a molar excess of DTPA, cysteine or glutathione solutions at $37^{\circ}C$ for 1 hour. The radiolabeled liposomes were also incubated in the presence of human serum at $37^{\circ}C$ for 24 hours. Results: 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized with 77.3% overall yield. The HYNIC concentration in the PEG-coated liposome dispersion was 1.08 mM. In condition of considering the measured liposomal size of 106 nm, the phospholipid concentration of $77.5\;{\mu}mol/m{\ell}$ and the liposomal particle number of $5.2{\times}10^{14}$ liposomes/ml, it is corresponded to approximate 1,250 nicotinyl hydrazine group per liposome in HYNIC-PEG-liposome. The removal of free $^{99m}Tc$ was not necessary because the labeling efficiency were above 99%. The radiolabeled liposomes maintained 98%, 96% and 99%, respectively, of radioactivity after incubation with transchelators. The radiolabeled liposomes possessed above 90% of the radioactivity in serum. Conclusion: These results suggest that the HYNIC can be synthesized easily and applied in labelling of PEG-liposomes with $^{99m}Tc$.