Neuroprotective effects of fermented tea in MPTP-induced Parkinson's disease mouse model via MAPK signaling-mediated regulation of inflammation and antioxidant activity.

Parkinson's disease (PD) is a neurodegenerative disorder that is characterized by dopaminergic neuronal damage. In this study, three tea extracts from Hadong, Korea, were evaluated in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity damage model (C57BL/6 mice) for their therapeutic effects against PD: green tea (GT), semi-fermented tea (SFT), and fermented tea (FT). Theaflavin content in the teas increased but catechin content decreased with the degree of fermentation. In addition, SFT showed the highest theanine and γ-aminobutyric acid contents. SFT at a concentration of 25 µg/mL showed the highest activity in the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay among all samples. Furthermore, the 2,2'-azino-bis 3-ethylbenzothiazoline-6-sulfonic acid radical scavenging activity of 25 µg/mL SFT was higher than that of l-ascorbic acid. Fermented tea suppressed the expression of inflammatory cytokines, such as interleukin-6, tumor necrosis factor-alpha, inducible nitric oxide synthase, cyclooxygenase-2, and macrophage-1, as well as inhibited overexpression of apoptotic signals, including p-53, cleaved caspase-3, and poly (ADP-ribose) polymerase-1. Moreover, GT, SFT, and FT regulated the MPTP-induced oxidative stress-related factors, including superoxide dismutase, glutathione-S-transferase, and nicotinamide adenine dinucleotide phosphate oxidase 4. Fermented tea also alleviated MPTP-induced behavioral impairment and dopaminergic neuronal damage and reduced α-synuclein levels. These results indicate that fermented tea is effective for the treatment of neuro-inflammatory, neuro-apoptotic, and neuro-oxidative disorders.

Collagen Extract Derived from Yeonsan Ogye Chicken Increases Bone Microarchitecture by Suppressing the RANKL/OPG Ratio via the JNK Signaling Pathway.

Yeonsan Ogye is a traditional Korean chicken breed (Gallus domesticus, GD), with a dominant gene for fibromelanosis, showing entirely black fluffy head feathers, ear lobes, and pupils. GD collagen extract (78.6 g per 100 g total protein) was derived from the flesh of Yeonsan Ogye. The effects of GD collagen on bone mass, microarchitecture, osteogenic, osteoclastogenic differentiations, and function factor expression were investigated in ovariectomized (OVX) rats. GD collagen stimulated osteogenesis in OVX rats and increased tibial bone strength and calcium content. Micro-computed tomography analysis of tibia cross-sections revealed that GD collagen attenuated the OVX-induced changes in trabecular thickness, spacing, and number. GD collagen stimulated alkaline phosphatase activity, bone-specific matrix proteins (alkaline phosphatase (ALP), osteocalcin, collagen type I (COL-I)) and mineralization by activating bone morphogenetic protein 2 (BMP-2)/mothers against decapentaplegic homolog 5 (SMAD5)/runt-related transcription factor 2 (Runx2). GD collagen inhibited osteoclast differentiation and function gene markers (TRAP, cathepsin K) by interfering with the Wnt signaling, increasing OPG production, and reducing the expression of RANKL, TRAP, and cathepsin K. GD collagen promoted osteogenesis by activating the p38 signal pathway and prevented osteoclastogenesis by lowering the RANKL/OPG ratio and blocking the JNK signaling pathway. Dietary supplementation with GD collagen might inhibit osteoclastogenesis, stimulate osteoblastogenesis, and regulate bone metabolism.

Inhibitory Effect of Centella asiatica Extract on DNCB-Induced Atopic Dermatitis in HaCaT Cells and BALB/c Mice.

Atopic dermatitis (AD) is a chronic inflammatory skin disease caused mainly by immune dysregulation. This study explored the anti-inflammatory and immunomodulatory effects of the Centella asiatica ethanol extract (CA) on an AD-like dermal disorder. Treatment with CA inhibited the expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in a dose-dependent manner in inflammatory stimulated HaCaT cells by interferon-γ (IFN-γ) and TNF-α-triggered inflammation. Eight-week-old BALB/c mice treated with 2,4-dinitrochlorobenzene (DNCB) were used as a mouse model of AD. In AD induce model, we had two types treatment of CA; skin local administration (80 µg/cm2, AD+CA-80) and oral administration (200 mg/kg/d, AD+CA-200). Interestingly, the CA-treated groups exhibited considerably decreased mast cell infiltration in the ear tissue. In addition, the expression of IL-6 in mast cells, as well as the expression of various pathogenic cytokines, such as TNF-α, IL-4, IL-5, IL-6, IL-10, IL-17, iNOS, COX-2, and CXCL9, was reduced in both AD+CA-80 and AD+CA-200 groups. Collectively, our data demonstrate the pharmacological role and signaling mechanism of CA in the regulation of allergic inflammation of the skin, which supports our hypothesis that CA could potentially be developed as a therapeutic agent for AD.

Calcium Supplement Derived from Gallus gallus domesticus Promotes BMP-2/RUNX2/SMAD5 and Suppresses TRAP/RANK Expression through MAPK Signaling Activation.

The present study evaluated the effects of a calcium (Ca) supplement derived from Gallus gallus domesticus (GD) on breaking force, microarchitecture, osteogenic differentiation and osteoclast differentiation factor expression in vivo in Ca-deficient ovariectomized (OVX) rats. One percent of Ca supplement significantly improved Ca content and bone strength of the tibia. In micro-computed tomography analysis, 1% Ca supplement attenuated OVX- and low Ca-associated changes in bone mineral density, trabecular thickness, spacing and number. Moreover, 1% Ca-supplemented diet increased the expression of osteoblast differentiation marker genes, such as bone morphogenetic protein-2, Wnt3a, small mothers against decapentaplegic 1/5/8, runt-related transcription factor 2, osteocalcin and collagenase-1, while it decreased the expression of osteoclast differentiation genes, such as thrombospondin-related anonymous protein, cathepsin K and receptor activator of nuclear factor kappa B. Furthermore, 1% Ca-supplemented diet increased the levels of phosphorylated extracellular signal-regulated kinase and c-Jun N-terminal kinase. The increased expression of osteoblast differentiation marker genes and activation of mitogen-activated protein kinase signaling were associated with significant increases in trabecular bone volume, which plays an important role in the overall skeletal strength. Our results demonstrated that 1% Ca supplement inhibited osteoclastogenesis, stimulated osteoblastogenesis and restored bone loss in OVX rats.

Hederagenin Supplementation Alleviates the Pro-Inflammatory and Apoptotic Response to Alcohol in Rats.

In this study, we determined the effects of hederagenin isolated from Akebia quinata fruit on alcohol-induced hepatotoxicity in rats. Specifically, we investigated the hepatoprotective, anti-inflammatory, and anti-apoptotic effects of hederagenin, as well as the role of AKT and mitogen-activated protein kinase (MAPK) signaling pathways in ethanol-induced liver injury. Experimental animals were randomly divided into three groups: normal (sham), 25% ethanol, and 25% ethanol + hederagenin (50 mg/kg/day). Each group was orally administered the respective treatments once per day for 21 days. Acetaldehyde dehydrogenase-2 mRNA expression was higher and alcohol dehydrogenase mRNA expression was lower in the ethanol + hederagenin group than those in the ethanol group. Pro-inflammatory cytokines, including TNF-α, IL-6, and cyclooxygenase-2, significantly increased in the ethanol group, but these increases were attenuated by hederagenin. Moreover, Western blot analysis showed increased expression of the apoptosis-associated protein, Bcl-2, and decreased expression of Bax and p53 after treatment with hederagenin. Hederagenin treatment attenuated ethanol-induced increases in activated p38 MAPK and increased the levels of phosphorylated AKT and ERK. Hederagenin alleviated ethanol-induced liver damage through anti-inflammatory and anti-apoptotic activities. These results suggest that hederagenin is a potential candidate for preventing alcoholic liver injury.

Radical Scavenging Activities of Tannin Extracted from Amaranth (Amaranthus caudatus L.).

This study investigates the bioactivity of tannin from amaranth (Amaranthus caudatus L.) extracts. The antioxidant activities of the extracts from amaranth leaves, flowers, and seeds were evaluated. Tannin from leaves of amaranth has been evaluated for superoxide scavenging activity by using DPPH and ABTS(+) analysis, reducing power, protective effect against H2O2-induced oxidative damage in L-132 and BNL-CL2 cells, and inhibition of superoxide radical effects on HL-60 cells. At a concentration of 100 µg/ml, tannin showed protective effects and restored cell survival to 69.2% and 41.8% for L-132 and BNL-CL2 cells, respectively. Furthermore, at the same concentration, tannin inhibited 41% of the activity of the superoxide radical on HL-60 cells and 43.4% of the increase in nitric oxide levels in RAW 264.7 cells. The expression levels of the antioxidant-associated protein SOD-1 were significantly increased in a concentration-dependent manner in RAW 264.7 cells treated with tannin from amaranth leaves. These results suggest that tannin from the leaves of Amaranthus caudatus L. is a promising source of antioxidant component that can be used as a food preservative or nutraceutical.

Effects of TGFbeta1 and extracts from Cervus korean TEMMINCK var. mantchuricus Swinhoe on acute and chronic arthritis in rats.

AIM OF THE STUDY: To elucidate the pharmacological activities of deer antler acupuncture and TGF61538;1 on the acute and chronic phases of rheumatoid arthritis diseases. MATERIALS AND METHODS: Polyarthritis rats were administered with TGF61538;1 and water extract of deer antler acupunture (DAA), prepared from the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe. TGF61538; (0.1 to 2 61549;g/animal) and DAA (5-100 61549;g/kg animal) were initiated 1 day before an arthritogenic dose of streptococcal cell wall fragments to see the effects on the joint swelling and distortion during the acute phase and the chronic phase of the disease. Arthritic index suppression of rat arthritis model was examined by TGF61538; and DAA administrations. RESULTS: TGF61538;1 and DAA diminished the polyarthritis development in rats. TGF61538; and DAA eliminated the joint swelling and distortion observed during the acute phase and the chronic phase of the disease. The TGF61538; and DAA suppressed the arthritis progress when administration was begun after acute phase of arthritis. DISCUSSION: Consistent with the inhibition of inflammatory cell recruitment into the synovium, TGF61538;1 and DAA reversed the leukocytosis associated with the chronic phase of the arthritis, respectively.

Enhancement of learning and memory by a medicinal formulation, Saenhyetang, in mice.

The effects on memory and learning ability of the Korean herbal medicine, Saenhyetang (SHT), which is consisted of nine herbs, were investigated. Hot water extracts (HWE-SHT) and ethanol extracts (EE-SHT) were used for the studies. It was shown that N-methyl-d-aspartate (NMDA) receptor 2B (NR2B) was increased in the forebrains of SHT-administrated mice (HWE-SHT), leading to enhanced activation of NMDA receptors, facilitating synaptic potentiation in response to stimulation at 10-100Hz. These HWE-SHT-treated mice exhibit superior ability in learning and memory in various behavioral tasks, showing that NR2B is enhanced by HWE-SHT treatment and also is critical in gating the age-dependent threshold for plasticity and memory formation. NMDA receptor-dependent modifications, which were mediated in part by HWE-SHT administration, of synaptic efficacy, therefore, represent a mechanism for associative learning and memory. Results suggest that oriental medical enhancement of NR2B attributes such as intelligence and memory in mammals is feasible. On the other hand, to examine the effects of EE-SHT on the learning and memory in experimental mice, the passive and active avoidance responses were studied. The EE-SHT ameliorated the memory retrieval deficit induced by ethanol, but not other memory impairment in mice. EE-SHT (10, 20mg/100g, p.o.) did not affect the passive avoidance responses of normal mice in the step through and step down tests, the conditioned and unconditioned avoidance responses of normal mice in the shuttle box and lever press performance tests, and the ambulatory activity of normal mice in normal condition. However, EE-SHT was shown to significantly decrease the spontaneous motor activity during the shuttle box test, and also to prolong the sleeping time induced by pentobarbital in mice at 20mg/kg. These results suggest that EE-SHT has an ameliorating effect on memory retrieval impairment and a weak tranquilizing action.

Immunosuppressive activity of deer antler extracts of Cervus korean TEMMINCK var. mantchuricus Swinhoe, on type II collagen-induced arthritis.

Unossified horn or pilose antler cut from deer, which belong to the Cervidae generally is termed Nokyong. Nokyong is one of the most famous Korean traditional medicines and has been considered to possess sexual-reinforcing and antiaging actions. In this study, water extract of deer antler extract (DAA) prepared from the growing antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe was used to investigate the efficacy of the DAA on the development of type II collagen (CII)-induced arthritis (CIA) in rats. Male rats were immunized with an emulsion of 200 microg of CII and complete Freund's adjuvant (CFA). The rats then were administered by injection a suspension of DAA or phosphate-buffered saline. The effect of DAA on cellular responses to CII was examined. The injection of DAA suppressed the CII-specific secretion of interferon (IFN)-gamma from splenocytes ex vivo. The influence of DAA also was evaluated on the incidence and development of arthritis in rat CIA. Rats were immunized twice at a 3-wk interval with bovine CII, with DAA being given by injection once a d for 14 d with four different regimens. A 14-d course of DAA treatment at a daily dose of 100 microg/kg, which began on the d of the first CII immunization, suppressed the development of arthritis, as well as antibody formation and delayed-type hypersensitivity to CII. Treatment with DAA resulted in inhibition of development of arthritis and immune responses to CII.

Effects of traditional herbal medicine, Hwaotang, on atherosclerosis using the spontaneous familial hypercholesterolemia model, Kurosawa and Kusanagi-hypercholesterolemic rabbits and the venous thrombosis rats.

Hwaotang (HOT), a traditional Korean medicinal formulation, is a dried decoctum of a mixture of seven herbal medicines, consisting of Angelica gigantis Radix, Rehmanniae Radix, Paeoniae Radix, Ciniamomi Cortex, Cnidii Rhizoma, Persicae Semen and Carthami Flos. In the present study, the inhibitory effects and anti thrombic properties of HOT on the progression of atherosclerotic lesions were studied using the spontaneous familial hypercholesterolemia (FH) model, Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits and rats. Changes in blood chemistry, pathology and low-density lipoprotein (LDL) oxidation were measured in a control and HOT group. In the control group, the area of atheromatous plaques of the aorta progressed between week 12 (36.65%) and week 14 (46.22%). This progression of atherosclerotic lesions did not occur in the HOT-treated group after 12 (24.24%) and 14 (23.34%) weeks. Antioxidative effects on LDL were seen in the HOT in weeks 12 and 14. HOT improved the hypercholesterolemia in the KHC rabbits. On the other hand, HOT and five of the seven herbs, except Cnidii Rhizoma and Carthami Flos, inhibited the endotoxin-induced hepatic venous thrombosis in high cholesterol diet-treated rats. However, Ciniamomi Cortex showed a very weak inhibitory effect on the endotoxin-induced hepatic venous thrombosis. The extract also inhibited the endotoxin-induced decrease in blood platelets and fibrinogen, and endotoxin-induced increase in fibrin degradation products (FDP) on disseminated intravascular coagulation in normal rats. In conclusion, these results suggest that HOT has inhibitory effects on the development of atheromatous plaque formation in spontaneous FH rabbits. It is also suggested that the antioxidative effects of HOT on LDL led to the beneficial effects observed in this study. The protection by HOT and its herbs on the artificially induced ischemic infarction might be related to their inhibitory effects on disseminated intravascular coagulation, platelet coagulation and thrombotic action.

Neuroprotective effect of Chuk-Me-Sun-Dan on NMDA- and AMPA-evoked nitric oxide synthase activity in mouse brain.

Chukmesundan (CMSD) is composed of 8 medicinal herbs including Panex ginseng C.A. MEYER, Atractylodes macrocephala KOID, Poria cocos WOLF, Pinellia ternata BREIT, Brassica alba BOISS, Aconitum carmichaeli DEBX, Cynanchum atratum BGE, and Cuscuta chinensis LAM and used for the treatment of various symptoms accompanying hypertension and cerebrovascular disorders. This study was carried out to examine the effects of CMSD on N-methyl-D-aspartate (NMDA)-evoked, and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-evoked nitric oxide synthase (NOS) activity in mouse brain. In adult forebrain, CMSD influences neuronal maintenance and is neuroprotective in several injury models through mechanisms that are incompletely understood. Interaction is observed between CMSD and nitric oxide (NO). Because NO affects both neural plasticity and degeneration, we hypothesized that CMSD might rapidly modulate NO production. Using in vivo microdialysis we measured conversion of L-[14C] arginine to L-[14C] citrulline as an accurate reflection of NOS activity in adult mouse hippocampus. CMSD significantly reduced NOS activities to 62% of basal levels within 2 days of onset of delivery and maintained NOS activity at less than 45% of baseline throughout 3 days of delivery. These effects did not occur with control (distilled water) and were not mediated by effect of CMSD on glutamate levels. In addition, simultaneous delivery of CMSD treatment prevented significant increases in NOS activity triggered by the glutamate receptor agonists NMDA and AMPA. Rapid suppression by CMSD of basal and glutamate-stimulated NOS activity may regulate neuromodulatory functions of NO or protect neurons from NO toxicity and suggests a novel mechanism for rapidly mediating functions of CMSD. It is shown that NMDA receptor stimulation leads to activation of p21ras (Ras) through generation of NO via neuronal NOS. The competitive NOS inhibitor, L-nitroarginine methyl ester, and CMSD prevents Ras activation elicited by NMDA, thus supporting the physiologic relevance of endogenous NO regulation of Ras. These results suggest that Ras is a physiologic target of endogenously produced NO and indicates a signaling pathway for NMDA receptor activation that may be important for long-lasting neuronal responses.

Inhibitory effect of the Korean herbal medicine, Dae-Jo-Whan, on platelet-activating factor-induced platelet aggregation.

The anti-thrombic properties of the Korean herbal medicine, Dae-Jo-Hwan (DJW) were investigated. Water extracts, a 70% methanol (MeOH) extract and an ethyl acetate (EtOAc) soluble fraction (III) from DJW inhibited platelet-activating factor (PAF)-induced platelet aggregation in vitro and in vivo assays. The extracts of DJW and eleven herbs from which it is derived, except for Panax ginseng Meyer, Angelica sinensis (OLIV.) DIELS and Schisandra chinensis Baill., inhibited AA-induced blood platelet aggregation to various extents. The effects observed with total DJW was synergistic over-additive rather that additive since the sum of single contributions was lower than the effect of the total extract. Fraction III was specially protected against the lethality of PAF, while verapamil did not afford any protection. Exogenously applied arachidonic acid (AA) (100 microM) led to a 89% platelet aggregation, the release of 14 pmol of ATP, and the formation of either 225 pg of thromboxane A2 (TXA2) or 45 pg of prostaglandin E2 (PGE2), each parameter being related to 10(6) platelets. An application of DJW 5 min before AA, dose-dependently diminished aggregation, ATP-re lease, and the synthesis of TXA2 and PGE2, with IC(50) values of 70, 87, 65 and 72 microg/ml, respectively. The similarity of the IC(50) values suggests the inhibition of cyclooxygenase (COX) by DJW as the primary target, thus suppressing the generation of TXA2, which induces platelet aggregation and the exocytosis of ATP by its binding on TXA2-receptors. These results indicate that DJW shows anti-thrombotic action on human platelets and inhibits the action of PAF in vivo by an antagonistic effect on PAF. Therefore, it may be useful in treating disorders caused by PAF.

Caffeic acid phenyl ester in propolis is a strong inhibitor of matrix metalloproteinase-9 and invasion inhibitor: isolation and identification.

BACKGROUND: Propolis has been used as a folk medicine and has several proven biological activities. Herbal remedies recommended for cancer therapies in Korea. METHODS: Matrix metalloproteinase (MMP)-9-inhibitory activity of propolis has been assessed. CAPE as an acting compound was isolated and molecular structure was determined. Anti-invasion activity of CAPE was assayed using hepatocarcinoma cells. RESULTS: Propolis ethanol extracts showed a strong inhibitory effect of MMP-9 activity, which is known to be involved in tumor cell invasion and metastasis in a concentration-dependent manner on zymography. Assay guided fractionation led to the isolation of a caffeic acid phenyl ester (CAPE) as the compound responsible for the anti-MMP-9 activity. CAPE was obtained by reversed-phase HPLC, and its structure was elucidated by fast atom bombardment mass spectrometry and tandem mass spectrometry. The purified CAPE inhibited MMP-9 activity with the IC(50) of 1.0-2.0 nmol/l. CONCLUSIONS: CAPE possesses selective antiproliferative activity toward hepatocaricoma cell line Hep3B, but not primary cultured mouse hepatocytes.

Effect of bee venom on aromatase expression and activity in leukaemic FLG 29.1 and primary osteoblastic cells.

The effect of bee venom aqua-acupuncture (BVA) (api-toxin), a traditional immunosuppressive Korean aqua-acupuncture, on the bone function in human osteoblastic cells was studied. To provide insights into the effect of BVA on aromatase activity in bone-derived cells, we examined the human leukaemic cell line FLG 29.1, which is induced to differentiate toward the osteoclastic phenotype by TPA and TGF-beta1, and the primary first-passage osteoblastic cells (hOB). Southern blot of RT-PCR products with a 32P-labeled cDNA probe for the human aromatase demonstrated that FLG 29.1 and hOB cells express aromatase mRNA. Gene expression and enzyme activity were stimulated in a time-dependent fashion by 5.0 microl/ml BV and by either 1-50 nM TPA or 0.01-0.5 ng/ml TGF-beta1, with maximal responses after 2-3 h exposure. After 24 h incubation of the cells in the absence of these stimuli the aromatase mRNA and the protein were barely detectable. These findings demonstrate that cells of the osteoclastic lineage synthesize aromatase in vitro by the local cytokine of TGF-beta1 and BVA. These can offer an explanation for the lack of development of osteoarthritis in BVA-treated patients.

Zedoariae rhizoma and curcumin inhibits platelet-derived growth factor-induced proliferation of human hepatic myofibroblasts.

During the course of liver fibrogenesis, hepatic myofibroblast cells (hMF), mostly derived from hepatic stellate cells (HSC), proliferate and synthesize excessive amounts of extracellular matrix (ECM) components. To evaluate the antiproliferative effect of a traditional herbal medicine, Zedoariae rhizoma water extracts (ZR) was examined on the growth inhibition of human hMF since proliferation of hMF is known to be central for the development of fibrosis during liver injury, and factors that may limit their growth are potential antifibrotic agents. The aim of this study was to test the effects of ZR on the proliferation and to clarify the molecular mechanisms of ZR inhibition of HSC proliferation in cultured human hMF. The cells were stimulated by platelet-derived growth factor (PDGF)-BB in the presence or absence of ZR. Proliferation was determined by bromodeoxy-uridine (BrdU) incorporation. The mRNA expressions of collagen alpha1(I) and (IV) were evaluated by a quantitative reverse transcription-polymerase chain reaction (RT-PCR). PDGF-receptor tyrosine phosphorylation was detected using anti-phosphotyrosine antibody. PDGF-receptor radioligand binding assay was performed by [125I]PDGF-BB. ZR inhibited the PDGF-BB-induced cell-proliferation and collagen alpha1(I) and (IV) mRNA expressions. ZR reduced the autophosphorylation of the PDGF-receptor. ZR blocked PDGF-BB binding to its receptor in a non-competitive manner. Furthermore, the 80% aqueous acetone extract of ZR was also found to show a decreasing effect against the proportion of S phase cells after PDGF stimulation. To clarify the active compounds, the principal constituents of seven sesquiterpenes (curdione, dehydrocurdione, germacrone, curcumenol, isocurcumenol, zedoarondiol and curcumenone) and a diarylheptanoid (curcumin) were examined. Among them, curcumin was found to decrease the proportion of S phase cells after PDGF stimulation at a dose of 30-50 microM. Potent antiproliferative and antifibrogenic effects of ZR toward hMF indicated that ZR might have therapeutic implications in chronic liver disease, indicating a novel role for ZR as a growth inhibitory mediator and pointing out its potential involvement in the negative regulation of liver fibrogenesis. In conclusion, ZR has an inhibitory effect on PDGF-induced proliferation of hMF and the blocking of PDGF-BB binding to its receptor may be the mechanism behind this effect.

Suppressive effects of a water extract of Ulmus davidiana Planch (Ulmaceae) on collagen-induced arthritis in mice.

Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammatory and protective effects on damaged tissue, inflammation and bone resorption. Therefore, this study was undertaken to address (1) whether the water extract of the bark of Ulmus davidiana Planch (Ulmaceae) (UD) can modulate the expression of inducible inflammatory cytokines in mice; (2) in order to assess the therapeutic effects of UD in collagen-induced arthritis (CIA) in mice. DBA/1 mice were immunized with bovine type II collagen. After a second collagen immunization, mice were treated with UD orally at 100mg/kg once a day for 3 weeks. Paws were evaluated macroscopically for redness, swelling and deformities. The levels of TNF-alpha and IL-1beta in the ankle were examined. The severity of arthritis within the knee joints was evaluated by histological assessment of cartilage destruction and pannus formation. Administration of UD significantly suppressed the progression of CIA and inhibited the production of TNF-alpha and IL-1beta in the paws. The erosion of cartilage was dramatically reduced in mouse knees after treatment with UD. In the serum of UD-treated mice, the levels of IL-4 and IL-10, anti-inflammatory cytokines, were increased. From the results, it was concluded that administration of UD has therapeutic effects on CIA including protection of cartilage and RA for a potential therapy.

Stimulative effects of Drynariae Rhizoma extracts on the proliferation and differentiation of osteoblastic MC3T3-E1 cells.

Pharmacological factors are needed to prevent bone loss that occurs with increasing age. The chemical compounds that act on bone metabolism in herbal medicines, however, are poorly understood. Effects of traditional Korean medicine, Drynariae Rhizoma [Drynaria fortunei (kunze) J. Sm] extract (DR), on the osteoblastic proliferation and differentiation were investigated. The effect of DR, a natural phyto herb, on the proliferation and osteoblastic differentiation in non-transformed osteoblastic cells (MC3T3-E1) was studied. DR dose-dependently increased DNA synthesis (significant at 50-150 microg/ml). DR increased alkaline phosphatase (ALP) activity and prolyl hydroxylase activity of MC3T3-E1 cells (50-150 microg/ml). Antiestrogen tamoxifen eleminated the stimulation of proliferation and ALP activity of MC3T3-E1, which were induced by DR. DR at concentrations ranged from 30-100 microg/ml inhibited prostaglandin E2 production in MC3T3-E1. These results indicate that DR directly stimulates cell proliferation and differentiation of osteoblasts. These results also suggest and DR is effective for bone anti-resorptive action in bone cells.

Anti-bone resorption activity of deer antler aqua-acupunture, the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe (Nokyong) in adjuvant-induced arthritic rats.

Effect of deer antler aqua-acupunture (DAA), prepared from the pilose antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe, a traditional immunosuppressive acupuncture, was evaluated to assess the reductions in bone mass, strength, and turnover in adjuvant-induced arthritic rats. For measuring the above parameters, a 20-day dosing experiment was performed using 6-week-old female Lewis rats. Arthritis was induced by injecting the adjuvant into the hind paw of the Lewis rats. The age-dependent increases in the body weight, lumbar bone mineral content and density (BMC and BMD) and compressive strength were disturbed in the arthritic rats. At 10 days, the histomorphometric parameters of bone formation (BFR/BS and BFR/BV) and the serum osteocalcin levels were significantly reduced compared with the baseline controls of Lewis rats. However, the BMC values corrected for body weight did not differ significantly between the arthritic and normal rats, and the bone minerals were not reduced when they were compared with the baseline controls. At 20 days, the parameters of bone minerals and strength of the lumbar body in the arthritic rats, both with and without correction for body weight, were significantly reduced compared with the baseline controls. The trabecular mineralizing surface remained significantly reduced and the osteoclast numbers were increased. DAA at the doses of 10, 20, 50 and 100 microg/kg, administered by Shinsu (B23) acupuncture daily from the start of the experiment, significantly prevented the development of the chronic paw edema at 20 days. The reductions in the parameters such as bone minerals, strength, and trabecular bone formation, and the increase in osteoclast number were alleviated by this DAA. Age-dependent increases in the lumbar height, disturbed by the adjuvant injection, were also maintained. These results indicated that a 20-day-period is necessary to obtain sufficient reductions in the bone mass and strength of the lumbar body concerning the model of secondary osteoarthritis in adjuvant arthritic rats. DAA was able to prevent these reductions by modulating the bone turnover in this arthritis model.

Attenuating effect of a traditional korean formulation, Paeng-Jo-Yeon-Nyeon-Baek-Ja-In-Hwan (PJBH), on hydrogen peroxide-induced injury in PC12 cells.

The Paeng-Jo-Yeon-Nyeon-Baek-Ja-In-Hwan (PJBH) prescription is a dried decoctum consisting of a mixture of 18 medicinal herbs that include Semen Biotae, Fructus Torilis seu cnidii, Fructus Rubi, Herba Dendrobii, Radix Morindae officinalis, Cortex Eucommiae, Radix Aspragi, Radix Polygalae, Radix Dipsaci, Ramulus Cinnamomi, Rhizoma Acori graminei, Rhizoma Alismatis, Rhizoma Dioscoreae, Radix Ginseng, Radix Rehmanniae preparata, Fructus corni, Fructus Schisandrae and Herba Cistanches. The effect of PJBH extracts on H2O2-induced toxicity in the rat pheochromocytoma line PC12 was examined by measurements of cell lesion, level of lipid peroxidation and antioxidant enzyme activities, since free radicals are involved in neurodegeneration in Alzheimer's disease (AD). After a 30 min exposure of the cells to H2O2 (150 microM), a marked decrease in cell survival, activities of glutathione peroxidase and catalase as well as an increased production of malondialdehyde (MDA) were found. Pretreatment of the cells with PJBH (0.5-10 microg/ml) prior to H2O2 exposure significantly elevated cell survival, antioxidant enzyme activities and resulted in a decrease in the level of MDA. The effects of the PJBH on hydrogen peroxide-induced injury in PC12 cells were also examined. PJBH had a remarkable elevating effect on catalase and GSH-Px activities as well as cell survival, suggesting that cytoprotective effects of the PJBH are involved in stimulation against intermediate concentrations of H2O2-induced PC12 cell injury. The above-mentioned neuroprotective effects were also compared with the effect of tacrine. The results suggest that PJBH has potential for use as a novel neuronal therapeutic agent.

Inhibitory effects of deer antler aqua-acupuncture, the pilose antler of Cervus Korean TEMMINCK var mantchuricus Swinhoe, on type II collagen-induced arthritis in rats.

Water extract of deer antler aqua-acupunture (DAA) prepared from the growing antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe, was used to investigate the efficacy of a traditional immunosuppressive and immuno-activating Korean aqua-acupuncture, on the development of type II collagen (CII)-induced arthritis (CIA) in rats. The onset of arthritis was observed at the 24th day after the CII-immunization in rats, and the severity of CIA was gradually developed. As compared with rats treated with saline, DAA i.p. injected at doses of more than 50 microg/kg once a day for 14 days inhibited the ability of inguinal lymph node cells to produce T cell cytokines interleukin 2 and interferon-gamma when the cells were obtained from rats 24 days after immunization and cultured in vitro with CII. Treatment with DAA also inhibited the production of macrophage cytokines interleukin-1beta, IL-6 and tumor necrosis factor alpha in response to in vitro stimulation of lymph node and macrophage cells with CII. In addition, in order to evaluate the influence of DAA on the incidence and development of arthritis in rat CIA, rats were immunized twice at a 3-week interval with bovine CII, with DAA being given i.p. once a day for 14 days with four different regimens. A 14-day course of DAA treatment at a daily dose of 100 microg/kg, which began on the day of the first CII immunization, suppressed the development of arthritis, as well as antibody formation and delayed-type hypersensitivity to CII. Treatment with DAA, which started on the same day as the booster immunization, also resulted in inhibition of development of arthritis and of immune responses to CII. However, treatment with DAA, which was prophylactically started prior to a primary immunization, did not inhibit the development of arthritis and immune response to CII. Furthermore, DAA extract did not affect the established diseases.