KiHwan Bae

Cholesterol-Lowering Activity of Naringenin via Inhibition of 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase and Acyl Coenzyme A:Cholesterol Acyltransferase in Rats

The effects of dietary supplementation of a citrus bioflavonoid, naringenin, on the cholesterol metabolism were studied. For 42 days male rats were fed a 1% (wt/wt) high-cholesterol diet with or without a naringenin supplementation (0.1%, wt/wt) to study its effect on plasma lipid levels, hepatic lipid contents, activities of hepatic acyl coenzyme A:cholesterol O-acyltransferase (ACAT) and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, and the excretion of fecal neutral sterols. Naringenin did not significantly alter the concentration of plasma triglycerides, but lowered the plasma cholesterol (3.80 vs. 3.12 mmol/l) concentration and the hepatic cholesterol content (70.3 vs. 54.0 mg/g) significantly (p < 0.05) compared to those of the controls. HMG-CoA reductase (1,879.0 vs. 1,715.0 pmol/min/mg) and ACAT activities (806.0 vs. 563.0 pmol/min/mg) were significantly lower in the naringenin-supplemented group than in controls. Naringenin supplementation caused a marked decrease in the excretion of fecal neutral sterols (242.9 mg/day) compared to the controls (521.9 mg/day). These results show that naringenin lowers the plasma and hepatic cholesterol concentrations by suppressing HMG-CoA reductase and ACAT in rats fed a high-cholesterol diet.

Anti-inflammatory activity of flavonoids from Populus davidiana.

Anin vitro bioassay-guide revealed that the methanol (MeOH) extract of the stem bark ofPopulus davidiana showed considerable inhibitory activity against cyclooxygenase (COX-1, COX-2). Continuous phytochemical study of the MeOH extract of this plant led to the isolation of ten flavonoids; sakuranetin (1), rhamnocitrin (2), 7-O-methylaromadendrin (3), naringenin (4), eriodictyol (5), aromadendrin (6), kaempferol (7), neosakuranin (8), sakuranin (9) and sakurenetin-5,4′-di-β-D-glucopyranoside (10). Their structures were identified on the basis of their physicochemical and spectroscopic analyses. The isolated compounds,1–10, were tested for their inhibitory activities against COX-1 and COX-2. Compound7 was found to have potent inhibitory effect on COX-1 and a moderate effect on COX-2, meanwhile, compounds1–6 showed moderate inhibition against COX-1 only. Moreover, compounds5–8 exhibited suppressive effects on xanthine oxidase (XO). These results may explain, in part, the traditional uses ofP. davidiana in ethnomedicine.

Antioxidant activities of coumarins from Korean medicinal plants and their structure-activity relationships.

The aim of this work was to study the structure–activity relationships of the antioxidant activity of natural coumarins isolated from four Korean medicinal plants (1–17) and four purchased coumarins (18–21). The free radical scavenging and lipid peroxidation assays revealed that five phenolic coumarins, scopoletin (1), aesculetin (2), fraxetin (3), umbelliferone (18) and daphnetin (19), possessed considerable antioxidant activities. The coumarins having a catechol group, 2, 3 and 19, showed significant free radical scavenging activity and inhibitory effects on lipid peroxidation, indicating that the catechol group significantly contributed to the antioxidant activities of coumarins. In contrast, the sugar moiety markedly reduced the activities of coumarin glycosides. The results also demonstrate that the α‐pyrone ring of coumarins significantly enhanced the capacity of inhibiting oxidative reactions of coumarins. Copyright

Down-regulation of c-Src/EGFR-mediated signaling activation is involved in the honokiol-induced cell cycle arrest and apoptosis in MDA-MB-231 human breast cancer cells

Honokiol is a naturally occurring neolignan abundant in Magnoliae Cortex and has showed anti-proliferative and pro-apoptotic effects in a wide range of human cancer cells. However, the molecular mechanisms on the anti-proliferative activity in cancer cells have been poorly elucidated. In this study, we evaluated the growth inhibitory activity of honokiol in cultured estrogen receptor (ER)-negative MDA-MB-231 human breast cancer cells. Honokiol exerted anti-proliferative activity with the cell cycle arrest at the G0/G1 phase and sequential induction of apoptotic cell death in a concentration-dependent manner. The honokiol-induced cell cycle arrest was well correlated with the suppressive expression of CDK4, cyclin D1, CDK2, cyclin E, c-Myc, and phosphorylated retinoblastoma protein (pRb) at Ser780. Apoptosis caused by honokiol was also concomitant with the cleavage of caspases (caspase-3, -8, and -9) and Bid along with the suppressive expression of Bcl-2, but it was independent on the expression of Bax and p53. In addition, honokiol-treated cells exhibited the cleavage of poly (ADP-ribose) polymerase (PARP) and DNA fragmentation. In the analysis of signal transduction pathway, honokiol down-regulated the expression and phosphorylation of c-Src, epidermal growth factor receptor (EGFR), and Akt, and consequently led to the inactivation of mTOR and its downstream signal molecules including 4E-binding protein (4E-BP) and p70 S6 kinase. These findings suggest that honokiol-mediated inhibitory activity of cancer cell growth might be related with the cell cycle arrest and induction of apoptosis via modulating signal transduction pathways.

Chemical constituents of the root of Dystaenia takeshimana and their anti-inflammatory activity.

In our ongoing search for bioactive compounds originating from the endemic species in Korea, we found that the hexane and EtOAc fractions of the MeOH extract from the root ofDystaenia takeshimana (Nakai) Kitagawa (Umbelliferae) showed cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) dual inhibitory activity by assessing their effects on the production of prostaglandin D2 (PGD2) and leukotriene C4 (LTC4) in mouse bone marrow-derived mast cells. By activity-guided fractionation, five coumarins, viz. psoralen (2), xanthotoxin (3), scopoletin (4), umbelliferone (5), and (+)-marmesin (6), together with β-sitosterol (1), were isolated from the hexane fraction, and two phenethyl alcohol derivatives, viz. 2-methoxy-2-(4′-hydroxyphenyl)ethanol (7) and 2-hydroxy-2-(4′-hydroxyphenyl)ethanol (8), three flavonoids, viz. apigenin (9), luteolin (10), and cynaroside (11), as well as daucosterol (12) were isolated from the EtOAc fraction using silica gel column chromatography. In addition, D-mannitol (13) was isolated from the BuOH fraction by recrystallization. Two of the coumarins, scopoletin (4) and (+)-marmesin (6), the two phenethyl alcohol derivatives (7, 8) and the three flavonoids (9–11) were isolated for the first time from this plant. Among the compounds isolated from this plant, the five coumarins as well as the three flavonoids showed COX-2/5-LOX dual inhibitory activity. These results suggest that the anti-inflammatory activity ofD. takeshimana might in part occurvia the inhibition of the generation of eicosanoids.

Vitexin, orientin and other flavonoids from Spirodela polyrhiza inhibit adipogenesis in 3T3‐L1 cells

To investigate the adipogenesis inhibitory effect on lipid accumulation, 3T3‐L1 cells were treated with fractions and isolated flavonoids of Spirodela polyrhiza. An ethanol extract of S. polyrhiza was fractionated into three fractions. The butanol soluble fraction (SPB) exhibited potent antiadipogenesis activity and decreased C/EBPα and PPARγ protein expression level in 3T3‐L1 cells without significant cytotoxicity. The flavonoids were isolated from SPB and their chemical structures were identified as chrysoeriol (1), apigenin (2), luteolin (3), vitexin (4), cosmosin (5), orientin (6) and luteolin‐7‐O‐β‐d‐glucoside (7) by spectroscopic analysis. Studies on the adipogenesis and intracellular triglyceride accumulation inhibitory effect showed that compounds 4 and 6 had the highest activity and decreased C/EBPα and PPARγ protein expression level in 3T3‐L1 cells. These results suggest that the flavonoids isolated from SPB, especially compounds 4 and 6, contribute to the inhibitory activity of S. polyrhiza in 3T3‐L1 cells. Copyright

Antioxidant and lipoxygenase inhibitory activity of oligostilbenes from the leaf and stem of Vitis amurensis.

ETHNOPHARMACOLOGICAL RELEVANCE The root and stem of Vitis amurensis (Vitaceae) have popularly used as traditional medicine for treatment of cancer and various pains in Korea and Japan. Recent studies, its root and stem possess anti-inflammatory, anti-tumor activities, and protective effects against beta-amyloid-induced oxidative stress. AIM OF THE STUDY This study deals with the isolation, structural identification of the potent bioactive compounds from the leaf and stem, and their antioxidant capacity, as well as anti-inflammatory effect via lipoxygenase inhibitory assay. MATERIALS AND METHODS All isolated compounds yielded after using column chromatography were identified base on the physico-chemical properties and 1D, 2D NMR spectra. The scavenge ability against DPPH and ABTS(+) radicals, and to inhibit lipid peroxidation, as well as lipoxygenase type I inhibitory activity of all isolates were performed using in vitro assays. RESULTS Eleven resveratrol derivatives (1-11), including a new oligostilbene cis-amurensin B (9), whose structures were determined on the basis of extensively spectral analyses, were isolated from the leaf and stem of Vitis amurensis. The isolates (1-11) were examined for their antioxidant activities by evaluating scavenge ability against DPPH and ABTS(+) radicals, and to inhibit lipid peroxidation. Stilbenes 1 and 4, and oligostilbenes 5-10 displayed moderate anti-lipid peroxidation activities, but all the isolates exhibited strong ABTS(+) radical scavenging activity in the dose-dependent manner. In addition, the isolates showed stronger inhibitory capacity against soybean lipoxygenase type I than that of baicalein, a positive control. Of the isolates, r-2-viniferin (8) exhibited the strongest scavenging activity against ABTS(+) radical with TEAC value of 5.57, and the most potential inhibitory effect on soybean lipoxygenase with the IC(50) value of 6.39 microM. CONCLUSION This is the first report on the potential antioxidant and LOX-1 inhibitory effects of oligostilbenes isolated from the leaf and stem of Vitis amurensis. In addition, chemical compositions isolated from the leaf and stem are almost similar to those isolated from the root of Vitis amurensis. Therefore, the results may explain, in part, the uses of the leaf and stem, as well as the root of Vitis amurensis in the Korean traditional medicine.

Stilbenes and oligostilbenes from leaf and stem of Vitis amurensis and their cytotoxic activity

Chromatographic separation of the EtOAc fraction from the leaf and stem of Vitis amurensis led to the isolation of six oligostilbenoids (i.e., r-2-viniferin (1), trans-amurensin B (2), trans-ɛ-viniferin (3), gnetin H (4), amurensin G (5), (+)-ampelopsin A (8)) and four stilbenoids (i.e., trans-resveratrol (6), (+)-ampelopsin F (7), piceatannol (9), and trans-piceid (10)). The structures have been identified on the basis of spectroscopic evidence and physicochemical properties. The isolates were investigated for cytotoxic activity against three cancer cell lines in vitro using the MTT assay method. Amurensin G (5) and trans-resveratrol (6) showed significant cytotoxic activity against L1210, K562 and HTC116 cancer cell lines with IC50 values ranging from 15.7 ± 2.1 to 30.9 ± 1.8 μM. (+)-Ampelopsin A (8) and trans-piceid (10) exhibited considerable cytotoxic activity against L1210 (IC50 values of 30.6 ± 4.1 and 28.7 ± 2.81 μM, respectively) and K562 (IC50 values of 38.6 ± 0.82 and 24.6 ± 0.76 μM, respectively). Gnetin H (4) showed only weak cytotoxic activity against L1210 with an IC50 value of 40.1 ± 4.23 μM. On the other hand, r-2-viniverin (1), trans-amurensin B (2), trans-ɛ-viniferin (3), (+)-ampelopsin F (7), and piceatannol (9) exhibited no activity on three cancer cell lines.

Discrimination of cinnamon bark and cinnamon twig samples sourced from various countries using HPLC-based fingerprint analysis

A simple and efficient HPLC method was developed to evaluate the quality of traditional herbal medicines made from cinnamon bark (CB) and cinnamon twig (CT). Seven major bioactive ingredients in 56 samples (24 CB and 32 CT) collected from China, Vietnam, and Indonesia were separated and quantified. The method was validated following the International Conference on Harmonisation (ICH) guidelines. A fingerprint analysis method to discriminate between CB and CT using major component content levels was developed. The discrimination process included the use of similarity indices and partial least-squares discriminant analysis (PLS-DA). Classification accuracy by the PLS-DA method was about 98%. The pattern analysis method was specific and could be readily used for the comprehensive evaluation of cinnamon samples. Therefore, an HPLC fingerprint in combination with pattern analysis provides a very flexible and reliable method for quality assessment of herbal drugs.

Cholinesterase inhibitory and anti-amnesic activity of alkaloids from Corydalis turtschaninovii.

In the course of screening plants used in Korean folk medicine as memory enhancers, a 70% ethanol extract of tuber from Corydalis turtschaninovii Besser (Papaveraceae) showed significant acetylcholinesterase (AChE) inhibitory activity. Repeated column chromatography led to the isolation of a new aporphine alkaloid, oxoglaucidaline (9), and a new protoberberine, pseudodehydrocorydaline (13) together with 14 known compounds (1-8, 10-12, and 14-16). The chemical structures of isolated compounds were elucidated base on extensive 1D and 2D NMR spectroscopic data. Compounds 1-16 were investigated in vitro for their anti-cholinesterase activity using the mice cortex AChE enzyme. In further study, the anti-amnesic activities of pseudoberberine (16) in mice on the learning and memory impairments induced by scopolamine (1.0 mg/kg, i.p.) were examined. This alkaloid (5.0 mg/kg, p.o.) administration significantly reversed cognitive impairments in mice by passive avoidance test (P<0.05). It also reduced escape latencies in training trials and prolonged swimming times in the target quadrant during the probe trial in the water maze task (P<0.05). These results indicated that Corydalis turtschaninovii due to its alkaloids have anti-cholinesterase activity and pseudoberberine and other alkaloids have anti-amnesic activities that may be useful for cognitive impairment treatment.