Comparison between the International Classification of Primary Care and the International Classification of Diseases Classifications in Primary Care in Korea.

BACKGROUND: The International Classification of Primary Care-2 (ICPC-2) is a classification method designed for primary care. Although previous studies have found that ICPC-2 is a useful tool for demonstrating the relationship between patients' expectations and health providers' diagnoses, its utility of ICPC-2 has yet to be fully studied in Korea. This study aimed to evaluate the practicality of ICPC-2 in Korean primary care. METHODS: The study was conducted at primary care clinics in Seoul and Gyeonggi areas from October to November 2015. Third-year family medicine residents examined and analyzed the medical records of patients who visited primary care physicians using ICPC-2, and the results were compared with those obtained using the International Classification of Diseases-10 (ICD-10) (Korean version: Korean Standard Classification of Diseases-7). RESULTS: A total of 26 primary care physicians from 23 primary care clinics participated in the study. Furthermore, 2,458 ICD-10 codes and 6,091 ICPC-2 codes were recorded from the data of 1,099 patients. The common disease codes were vasomotor and allergic rhinitis (J30), according to ICD-10, and acute upper respiratory infection (R74) in ICPC-2. Comparing disease status by body systems, the proportion of gastrointestinal disease with ICD-10 codes was significantly higher than that with ICPC-2 codes (P<0.001). Furthermore, patients with >4 diagnoses accounted for 36% of the ICD-10 classifications, whereas those with >4 diagnoses accounted for 4% of the ICPC-2 classifications. CONCLUSION: Introducing ICPC as a complementary means for diagnosing common diseases could be a practical approach in Korean primary care.

Association between Resting Heart Rate and Colorectal Cancer: Results from a Case-Controlled Study.

Previous studies evaluating associations between resting heart rate (RHR) and cancer-related mortality/prognosis have yielded conflicting results. We investigated whether elevations in RHR are associated with colorectal cancer (CRC). We conducted a case-controlled study involving 1241 CRC patients and 5909 cancer-free controls from the Korean National Health and Nutrition Examination Survey. After propensity score (PS) matching, 1207 CRC patients and 1207 matched controls were analyzed. Associations between RHR and CRC, colon, and rectal cancer were analyzed in appropriate patient subgroups using multiple and conditional logistic regression. Receiver operating characteristics analysis yielded the optimal RHR cut-point to predict CRC. RHR was significantly higher in CRC, colon, and rectal cancer patients than in controls (72.7 bpm in CRC, 72.8 bpm in colon cancer, 72.3 bpm in rectal cancer, and 68.7 bpm in controls; all p < 0.001). Analysis of data prior to PS matching yielded the following odds ratios (ORs) per RHR increment for CRC, colon, and rectal cancer: 1.043 (95% confidence intervals (CIs): 1.036-1.049), 1.045 (95% CI: 1.037-1.053), and 1.040 (95% CI: 1.030-1.051), respectively, in unadjusted models, and 1.043 (95% CI: 1.034-1.051), 1.046 (95% CI: 1.037-1.055), and 1.040 (95% CI: 1.027-1.052), respectively, in multivariable adjusted models. Patients with CRC, colon, and rectal cancer have a significantly higher RHR compared to cancer-free controls.

Relationship Between Handgrip Strength and Pulmonary Function in Apparently Healthy Older Women.

OBJECTIVES: To investigate the relationship between handgrip strength and pulmonary function. DESIGN: Cross-sectional study of a representative sample of older Korean women. SETTING: The Korean National Health and Nutrition Examination Survey. PARTICIPANTS: Community-dwelling women aged 65 and older without chronic diseases or pulmonary disease (N=605). MEASUREMENTS: Handgrip strength was measured using a digital hand dynamometer, and pulmonary function was tested according to guidelines of the American Thoracic Society/European Respiratory Society using a spirometry system. Impaired pulmonary function was defined as a lower limit of normal (LLN) or less of forced vital capacity (FVC) and forced expiratory volume in 1 second (FEV1). Odds ratios (ORs) and 95% confidence intervals (CIs) for impaired pulmonary function according to handgrip strength quartile were calculated using multiple logistic regression analysis. RESULTS: Mean FVC and FEV1 gradually increased in accordance with handgrip strength quartiles (all P <.001). After adjusting for age, body mass index, smoking status, alcohol ingestion, aerobic physical activity, resistance exercise, household income, and education level the odds of impaired pulmonary function were greater for participants in the first quartile of handgrip strength (≤19.25 kg) than for those in the fourth quartile (25.31-37.30 kg) (FVC LLN: OR=3.46, 95 % CI=1.52-7.88; FEV1 LLN: OR=2.62, 95 % CI=1.12-6.15). CONCLUSION: Handgrip strength was positively associated with pulmonary function in a dose-dependent manner. Given the health implications of pulmonary function, timely detection of weaker handgrip strength in older people may be useful in assessing potential pulmonary function impairment.

Korean Red Ginseng Protects Against Mitochondrial Damage and Intracellular Inflammation in an Animal Model of Type 2 Diabetes Mellitus.

Korean red ginseng (KRG), a heat-processed Korean ginseng (Panax ginseng C.A. Meyer), has been used as a traditional medicine for its beneficial effects on hyperglycemia. This study aimed to investigate whether the antidiabetic action of KRG in an animal model of type 2 diabetes mellitus (DM) is partly mediated by prevention of mitochondrial dysfunction and intracellular inflammation. Four-week-old C57BL/KsJ db/db mice (a genetic animal model of obese type 2 DM) and C57BL/KsJ db/+ mice were divided into three groups: db/+ mice (normoglycemic control group, n = 8), db/db mice (untreated DM group, n = 8), and db/db mice with KRG administration (KRG-treated DM group, n = 8). After 12 weeks, metabolic parameters of fasting blood glucose concentrations, hemoglobin A1c (HbA1c) level, insulin level, lipid profile, and leukocyte count were determined using high-performance liquid chromatography. Mitochondrial DNA (mtDNA) copy number and inflammatory marker (interleukin-6, cyclooxygenase-2, and C-reactive protein) expression levels were measured in skeletal muscle tissue using quantitative real-time PCR analysis. After 12 weeks of KRG treatment at 100 mg/kg, the fasting glucose, HbA1c, insulin, and low-density lipoprotein cholesterol concentrations were lower, whereas mtDNA copy numbers were higher in the KRG-treated DM group than in the untreated DM group. Compared with the untreated DM group, the messenger RNA expression levels of mitochondrial biogenesis-related transcription factors and inflammatory markers were lower in the KRG-treated DM group. In conclusion, KRG had a beneficial effect on the metabolic profile by preserving mitochondrial function and protecting against intracellular inflammation.

3'-Sialyllactose as an inhibitor of p65 phosphorylation ameliorates the progression of experimental rheumatoid arthritis.

BACKGROUND AND PURPOSE: 3'-Sialyllactose (3'-SL) is a safe compound that is present in high levels in human milk. Although it has anti-inflammatory properties and supports immune homeostasis, its effect on collagen-induced arthritis (CIA) is unknown. In this study, we investigated the prophylactic and therapeutic effect of 3'-SL on the progression of rheumatoid arthritis (RA) in in vitro and in vivo models. EXPERIMENTAL APPROACH: The anti-arthritic effect of 3'-SL was analysed with fibroblast-like synoviocytes in vitro and an in vivo mouse model of CIA. RT-PCR, Western blotting and ELISA were performed to evaluate its effects in vitro. Histological analysis of ankle and knee joints of mice with CIA was performed using immunohistochemistry, as well as safranin-O and haematoxylin staining. KEY RESULTS: 3'-SL markedly alleviated the severity of CIA in the mice by reducing paw swelling, clinical scores, incidence rate, serum levels of inflammatory cytokines and autoantibody production. Moreover, 3'-SL reduced synovitis and pannus formation and suppressed cartilage destruction by blocking secretion of chemokines, pro-inflammatory cytokines, matrix metalloproteinases and osteoclastogenesis via NF-κB signalling. Notably, phosphorylation of p65, which is a key protein in the NF-κB signalling pathway, was totally blocked by 3'-SL in the RA models. CONCLUSIONS AND IMPLICATIONS: 3'-SL ameliorated pathogenesis of CIA by suppressing catabolic factor expression, proliferation of inflammatory immune cells and osteoclastogenesis. These effects were mediated via blockade of the NF-κB signalling pathway. Therefore, 3'-SL exerted prophylactic and therapeutic effects and could be a novel therapeutic agent for the treatment of RA.

Serum ferritin level is positively associated with insulin resistance and metabolic syndrome in postmenopausal women: A nationwide population-based study.

OBJECTIVE: Serum ferritin, a marker of iron metabolism, has recently emerged as a biomarker of chronic low-grade inflammation. After menopause, there is a remarkable increase in insulin resistance (IR) and metabolic syndrome (MetS), which is increasingly being viewed as an inflammatory disease. Thus, we examined the associations of serum ferritin with insulin resistance and MetS in postmenopausal women. METHODS: A nationwide cross-sectional study was conducted to examine the relationship between serum ferritin and IR and MetS in 2734 postmenopausal women using data from the 2010-2012 Korean National Health and Nutrition Examination Survey. The odds ratios (ORs) and 95% confidence intervals (CIs) for insulin resistance (HOMA-IR≥75th percentile, 3.04) and MetS were calculated using multiple logistic regression analyses across serum ferritin quartiles (Q1,≤36.25; Q2, 36.56-56.56; Q3, 56.57-85.98; and Q4≥85.99ng/ml). RESULTS: The mean values of most cardiometabolic variables tended to increase proportionally with serum ferritin quartiles. The proportion of women with IR and MetS significantly increased in accordance with serum ferritin quartiles. Compared to individuals in the lowest quartile, the ORs (95% CIs) in the highest quartile were 2.06 (1.23-3.45) for IR and 1.92 (1.44-2.55) for MetS after adjusting for age, cigarette smoking, alcohol intake, and regular exercise. CONCLUSION: Serum ferritin levels were positively and independently associated with IR and MetS in postmenopausal women. These findings suggest that serum ferritin level in postmenopausal women may help to identify the presence of IR and MetS.

Factors Positively Influencing Health Are Associated with a Lower Risk of Development of Metabolic Syndrome in Korean Men: The 2007-2009 Korean National Health and Nutrition Examination Survey.

BACKGROUND: The prevalence of metabolic syndrome (MetS) has risen rapidly worldwide, including in South Korea. Factors related to lifestyle are closely associated with the development of MetS. The aim of this study was to investigate the association between MetS and a number of factors positively influencing health, namely non-smoking, low-risk drinking, sufficient sleep, regular exercise, and the habit of reading food labels, among Korean men. METHODS: This cross-sectional study included 3,869 men from the 2007-2009 Korean National Health and Nutrition Examination Survey. Information on five factors positively influencing their health was obtained using a self-reported questionnaire. We categorized subjects into four groups, depending on the number of positive factors reported (group I, 0-1 factor; group II, 2 factors; group III, 3 factors; group IV, 4-5 factors). RESULTS: Men who reported a greater number of positive health factors had better laboratory and anthropometric values than men who reported fewer positive health factors. The prevalence of MetS was 29.1, 27.2, 20.7, and 14.6% in groups I to IV, respectively. Compared to group I, odds ratios (95% confidence intervals) for MetS were 0.96 (0.78-1.19) in group II, 0.67 (0.52-0.87) in group III, and 0.52 (0.35-0.76) in group IV, after adjusting for confounding factors. Odds ratios for abdominal obesity, glucose intolerance, and hypertriglyceridemia were statistically significant. CONCLUSION: A greater number of positive lifestyle factors influencing health were associated with a lower risk of developing MetS, in a nationally representative sample of Korean men.

Recombinant Adeno-Associated Virus Expressing Truncated IK Cytokine Diminishes the Symptoms of Inflammatory Arthritis.

IK can downregulate interferon-gamma-induced major histocompatibility complex (MHC) class II expression through the MHC class II transactivator, which suggests that IK can inhibit the interactions between immune cells. We delivered adeno-associated virus serotype 2 (AAV2) encoding the genes for truncated IK (tIK) or green fluorescent protein (GFP) to DBA1/J mice via intravenous injection. Seven weeks after injection, collagen-induced arthritis was induced in the AAV2-treated mice. AAV2-tIK injection reduced the severity of arthritis and the percentage of pathogenic Th17 cells compared with AAV2-GFP injection. These results suggest a novel gene therapy strategy for treatment of inflammatory arthritis.

Antiplatelet activity of [5-(2-methoxy-5-chlorophenyl)furan-2-ylcarbonyl]guanidine (KR-32570), a novel sodium/hydrogen exchanger-1 and its mechanism of action.

The antiplatelet effects of a novel guanidine derivative, KR-32570 ([5-(2-methoxy-5-chlorophenyl) furan-2-ylcarbonyl]guanidine), were investigated with an emphasis on the mechanisms underlying its inhibition of collagen-induced platelet aggregation. KR-32570 significantly inhibited the aggregation of washed rabbit platelets induced by collagen (10 microg/mL), thrombin (0.05 U/mL), arachidonic acid (100 microM), a thromboxane (TX) A2 mimetic agent U46619 (9,11-dideoxy-9,11-methanoepoxy-prostaglandin F2, 1 microM) and a Ca2+ ATPase inhibitor thapsigargin (0.5 microM) (IC50 values: 13.8 +/- 1.8, 26.3 +/- 1.2, 8.5 +/- 0.9, 4.3 +/- 1.7 and 49.8 +/- 1.4 microM, respectively). KR-32570 inhibited the collagen-induced liberation of [3H]arachidonic acid from the platelets in a concentration dependent manner with complete inhibition being observed at 50 microM. The TXA2 synthase assay showed that KR-32570 also inhibited the conversion of the substrate PGH2 to TXB2 at all concentrations. Furthermore, KR-32570 significantly inhibited the [Ca2+]i mobilization induced by collagen at 50 microM, which is the concentration that completely inhibits platelet aggregation. KR-32570 also decreased the level of collagen (10 microg/mL)-induced secretion of serotonin from the dense-granule contents of platelets, and inhibited the NHE-1-mediated rabbit platelet swelling induced by intracellular acidification. These results suggest that the antiplatelet activity of KR-32570 against collagen-induced platelet aggregation is mediated mainly by inhibiting the release of arachidonic acid, TXA2 synthase, the mobilization of cytosolic Ca2+ and NHE-1.

Relationship between Blood Mercury Concentration and Bone Mineral Density in Korean Men in the 2008-2010 Korean National Health and Nutrition Examination Survey.

BACKGROUND: The results of previous studies on the association between blood mercury (Hg) and bone mineral density (BMD) are inconsistent. We therefore used a large-scale nationwide representative sample of Korean men to investigate the relationship between these two parameters. METHODS: A nationwide cross-sectional study was conducted using data from the 2008 to 2010 Korean National Health and Nutrition Examination Survey to evaluate the relationship between blood Hg and BMD and the prevalence of osteopenia or osteoporosis in 1,190 men over 50 years of age. BMD was measured by dual-energy X-ray absorptiometry. Osteopenia and osteoporosis were diagnosed for each body site according to World Health Organization T-score criteria. RESULTS: After adjusting for age, body mass index, caloric energy and calcium intake, vitamin D levels, fish consumption, alcohol consumption, smoking, and exercise, quartiles of blood Hg were positively associated with femur neck T-scores in multiple linear regression analysis (ß=0.06, P-value=0.03). Compared with the lowest blood Hg quartile, the odds ratios and 95% confidence intervals for diagnosis of osteopenia or osteoporosis in the second and fourth quartiles were 0.63 (0.41-0.99) and 0.57 (0.36-0.91), respectively, in the femur neck after adjusting for the same co-variables. CONCLUSION: High blood Hg levels were associated with reduced odds of decreased femur neck BMD in Korean men. However, subgroup analysis did not show a significant protective effect of blood Hg on osteoporotic fractures. Further research is necessary to clarify the association between blood Hg and BMD.

An antithrombotic agent, NQ301, inhibits thromboxane A2 receptor and synthase activity in rabbit platelets.

In previous studies we have reported that NQ301, a synthetic 1,4-naphthoquinone derivative, displays a potent antithrombotic activity, and that this might be due to antiplatelet effect, which was mediated by the inhibition of cytosolic Ca(2+) mobilization in activated platelets. In the present study, the effect of NQ301 on arachidonic acid cascade in activated platelets has been examined. NQ301 concentration-dependently inhibited washed rabbit platelet aggregation induced by collagen (10 microg/ml), arachidonic acid (100 microM) and U46619 (1 microM), a thromboxane A2 receptor agonist, with IC50 values of 0.60+/-0.02, 0.78+/-0.04 and 0.58+/-0.04 microM, respectively. NQ301 also produced a shift to the right of the concentration-effect curve of U46619, indicating a competitive type of antagonism on thromboxane A2/prostaglandin H2 receptor. NQ301 slightly inhibited collagen-induced arachidonic acid liberation. In addition, NQ301 potently suppressed thromboxane B2 formation by platelets that were exposed to arachidonic acid in a concentration-dependent manner, but had no effect on the production of prostaglandin D2, indicating an inhibitory effect on thromboxane A2 synthase. This was supported by thromboxane A2 synthase activity assay that NQ301 concentration-dependently inhibited thromboxane B2 formation converted from prostaglandin H2. Moreover, NQ301 concentration-dependently inhibited 12-hydroxy-5,8,10,14-eicosatetraenoic acid formation by platelets that were exposed to arachidonic acid. Taken together, these results suggest that NQ301 has a potential to inhibit thromboxane A2 synthase activity with thromboxane A2/prostaglandin H2 receptor blockade, and modulate arachidonic acid liberation as well as 12-hydroxy-5,8,10,14-eicosatetraenoic acid formation in platelets. This may also be a convincing mechanism for the antithrombotic action of NQ301.

BMS-191095, a cardioselective mitochondrial K(ATP) opener, inhibits human platelet aggregation by opening mitochondrial K(ATP) channels.

We evaluated the antiplatelet effects of two classes of ATP-sensitive potassium channel openers (K(ATP) openers) on washed human platelets, and the study's emphasis was on the role of mitochondrial K(ATP) in platelet aggregation. Collagen-induced platelet aggregation was inhibited in a dose dependent manner by lemakalim and SKP-450, which are potent cardio-nonselective K(ATP) openers, and also by cardioselective BMS-180448 and BMS-191095 (IC50: 1,130, >1,500, 305.3 and 63.9 microM, respectively), but a significantly greater potency was noted for the cardioselective K(ATP) openers. The latter two K(ATP) openers also inhibited platelet aggregation induced by thrombin, another important blood-borne platelet activator, with similar rank order of potency (IC50: 498.0 and 104.8 microM for BMS-180448 and BMS-191095, respectively). The inhibitory effects of BMS-191095 on collagen-induced platelet aggregation were significantly blocked by a 30-min pretreatment of platelets with glyburide (1 microM) or sodium 5-hydroxydecanoate (5-HD, 100 microM), a nonselective and selective mitochondrial K(ATP) antagonist, respectively, at similar magnitudes; this indicates the role of mitochondrial K(ATP) in the antiplatelet activity of BMS-191095. However, glyburide and 5-HD had no effect when they were added to the platelet cuvette immediately prior to the addition of BMS-191095. These findings indicate that cardioselective mitochondrial K(ATP) openers like BMS-191095 are able to exert cardioprotective effects in cardiac ischemia/reperfusion injury via dual mechanisms directed at the inhibition of platelet aggregation and the protection of cardiomyocytes, and both these mechanisms are mediated by mitochondrial K(ATP).

Antiplatelet effect of green tea catechins: a possible mechanism through arachidonic acid pathway.

We have previously reported that green tea catechins (GTC) showed an antithrombotic activity, which might be due to antiplatelet effect rather than anticoagulation. The present study was performed to investigate the effect of GTC on the arachidonic acid (AA) metabolism in order to elucidate a possible antiplatelet mechanism. GTC inhibited the collagen-, AA- and U46619-induced rabbit platelet aggregation in vitro in a concentration-dependent manner, with IC50 values of 61.0+/-2.5, 105.0+/-4.9 and 67.0+/-3.2 microg/ml, respectively. Moreover, GTC administered orally into rats inhibited the AA-induced platelet aggregation ex vivo by 46.9+/-6.1% and 95.4+/-2.2% at the doses of 25 and 50 mg/kg, respectively. [3H]AA liberation induced by collagen in [3H]AA incorporated rabbit platelets was significantly suppressed by GTC compared to the control. GTC also significantly inhibited the thromboxane A2 (TXA2) and prostaglandin D2 (PGD2) generations induced by addition of AA in intact rabbit platelets. GTC significantly inhibited TXA2 synthase activity in a concentration-dependent manner. Moreover, adenosine triphosphate (ATP) release from dense granule was inhibited by GTC in washed platelets. These results suggest that the antiplatelet activity of GTC may be due to the inhibition of TXA2 formation through the inhibition of AA liberation and TXA2 synthase.

Antiplatelet and antithrombotic activities of CP201, a newly synthesized 1,4-naphthoquinone derivative.

The antiplatelet and antithrombotic activities of a newly synthesized CP201, 2-(3,5-di-tert-butyl-4-hydroxyl)-3-chloro-1,4-naphthoquinone on human platelet aggregation in vitro and murine pulmonary thrombosis in vivo were examined. In addition, the antiplatelet activity of CP201 involved in calcium-signaling cascade was also investigated. CP201 showed concentration-dependent inhibitory effects on platelet aggregation induced by collagen and thrombin, with IC50 values of 4.1+/-0.3 and 4.6+/-0.4 microM, respectively. Orally administered CP201 protected mice against the collagen plus epinephrine-induced thromboembolic death in a dose-dependent manner. On the other hand, CP201 did not alter such coagulation parameters as activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) in human plasma in vitro. These results suggest that the antithrombotic activity of CP201 may be due to antiplatelet rather than anticoagulation activity. CP201 potently inhibited platelet aggregation challenged by calcium ionophore A23187 and thapsigargin, which is a selective inhibitor of the Ca(2+)-ATPase pump, in a concentration-dependent manner, indicating that CP201 may have an inhibitory effect on calcium-signaling cascade. This was supported by measuring [Ca2+]i in platelets loaded with fura-3AM, where CP201 inhibited the rise in cytosolic Ca2+ mediated by thrombin. Taken together, these results suggest that CP201 may be a promising antithrombotic agent, and the antithrombotic effect of CP201 may be due to antiplatelet activity, which was mediated, at least partly, by the inhibition of cytosolic calcium mobilization.

Antiplatelet activity of epigallocatechin gallate is mediated by the inhibition of PLCgamma2 phosphorylation, elevation of PGD2 production, and maintaining calcium-ATPase activity.

We have previously reported that green tea catechins displayed a potent antithrombotic effect by inhibition of platelet aggregation. In the present study, the antiplatelet and antithrombotic activities of epigallocatechin gallate (EGCG), the major catechin derived from green tea, were extensively investigated. EGCG inhibited arterial thrombus formation and U46619-, collagen-, and arachidonic acid (AA)-induced washed rabbit platelet aggregation in a concentration-dependent manner, with IC50 values of 61 +/- 3, 85 +/- 4, and 99 +/- 4 microM, respectively. In line with the inhibition of collagen-induced platelet aggregation, EGCG revealed blocking of the collagen-mediated phospholipase (PL) Cgamma2 and protein tyrosine phosphorylation, and it caused concentration-dependent decreases of cytosolic calcium mobilization, AA liberation, and serotonin secretion. In addition, the platelet aggregation, intracellular Ca2+ mobilization, and protein tyrosine phosphorylation induced by thapsigargin, a Ca2(+)-ATPase pump inhibitor, were completely blocked by EGCG. Contrary to the inhibition of AA-induced platelet aggregation, EGCG failed to inhibit cyclooxygenase and thromboxane (TX) A2 synthase activities, but it concentration-dependently elevated AA-mediated PGD2 formation. In contrast, epigallocatechin (EGC), a structural analogue of EGCG lacking a galloyl group in the 3' position, slightly inhibited collagen-stimulated cytosolic calcium mobilization, but failed to affect other signal transductions as did EGCG in activated platelets and arterial thrombus formation. These results suggest that antiplatelet activity of EGCG may be attributable to its modulation of multiple cellular targets, such as inhibitions of PLCgamma2, protein tyrosine phosphorylation and AA liberation, and elevation of cellular PGD2 levels, as well as maintaining Ca2(+)-ATPase activity, which may underlie its beneficial effect on the atherothrombotic diseases.

Inhibition of PDGF beta-receptor tyrosine phosphorylation and its downstream intracellular signal transduction in rat aortic vascular smooth muscle cells by kaempferol.

Kaempferol, a flavonoid present in human diet and plants, has been known to show cardiovascular protection via its anti-oxidant activity. In this study, we have investigated the effect of kaempferol on the proliferation of primary cultured rat aortic vascular smooth muscle cells (VSMCs). Kaempferol significantly inhibited 50 ng/mL platelet-derived growth factor (PDGF)-BB-induced proliferation and [3H]-thymidine incorporation into DNA at concentrations of 5, 20 and 50 microM without any cytotoxicity. Kaempferol also inhibited the c-fos mRNA expression induced by PDGF-BB concentration-dependently. In addition, consistent with the inhibition of cell proliferation and c-fos mRNA expression, kaempferol inhibited the PDGF beta-receptor (Rbeta) phosphorylation in a concentration-dependent manner. Accordingly, the downstream signal transductions of PDGF-Rbeta such as ERK1/2, Akt and PLC-gamma1 phosphorylation were also inhibited by kaempferol in the same pattern. These findings suggest that, in addition to its anti-oxidant activity, the cardiovascular protective effect of kaempferol may be mediated, at least in part, by the suppression of VSMC proliferation, which is due to the inhibition of PDGF-Rbeta tyrosine phosphorylation and its downstream intracellular signal transduction.

Antiplatelet activity of J78 (2-Chloro-3-[2'-bromo, 4'-fluoro-phenyl]-amino-8-hydroxy-1,4-naphthoquinone), an antithrombotic agent, is mediated by thromboxane (TX) A2 receptor blockade with TXA2 synthase inhibition and suppression of cytosolic Ca2+ mobilization.

We previously reported that J78 (2-chloro-3-[2'-bromo, 4'-fluoro-phenyl]-amino-8-hydroxy-1,4-naphthoquinone), a newly synthesized 1,4-naphthoquinone derivative, exhibited a potent antithrombotic effect, which might be due to antiplatelet rather than anticoagulation activity. In the present study, possible anti-platelet mechanism of J78 was investigated. J78 concentration-dependently inhibited rabbit platelet aggregation induced by collagen (10 microg/ml), thrombin (0.05 U/ml), arachidonic acid (100 microM), and U46619 (9,11-dideoxy-9,11-methanoepoxy-prostaglandin F(2); 1 microM), a thromboxane (TX) A(2) mimic, with IC(50) values of 0.32 +/- 0.01, 0.44 +/- 0.02, 0.50 +/- 0.04, and 0.36 +/- 0.02 microM, respectively. J78 also produced a shift to the right of the concentration-response curve of U46619, indicating an antagonistic effect on the TXA(2) receptor. J78 concentration-dependently inhibited collagen-induced arachidonic acid liberation. In addition, J78 potently suppressed TXA(2) formation by platelets that were exposed to arachidonic acid in a concentration-dependent manner but had no effect on the production of PGD(2), indicating an inhibitory effect on TXA(2) synthase. This was supported by a TXA(2) synthase activity assay that J78 concentration-dependently inhibited TXB(2) formation converted from PGH(2). Furthermore, J78 was also able to inhibit the [Ca(2+)](i) mobilization induced by collagen or thrombin at such a concentration that completely inhibited platelet aggregation. Taken together, these results suggest that the antiplatelet activity of J78 may be mediated by TXA(2) receptor blockade with TXA(2) synthase inhibition and suppression of cytosolic Ca(2+) mobilization.

Inhibitory effects of J78, a newly synthesized 1,4-naphthoquinone derivative, on experimental thrombosis and platelet aggregation.

Several compounds with the backbone of 1,4-naphthoquinone chemical structure have been reported to display antiplatelet and antithrombotic activities, indicating that this congener compound may be a new source in the antithrombotic drug development. In the present study, the possible antiplatelet activity and antithrombotic efficacy of J78 (2-chloro-3-[2'-bromo, 4'-fluoro- phenyl]-amino-8-hydroxy-1,4-naphthoquinone), a newly synthesized 1,4-naphthoquinone derivative, were examined. Orally administered J78 (50, 100 mg/kg) dose dependently protected mice against the collagen + epinephrine-induced thromboembolic death. Orally administered J78 also significantly inhibited the ADP- and collagen-induced rat platelet aggregation ex vivo, with inhibition values of 44 and 40%, respectively. J78 inhibited the collagen-, arachidonic acid- and thrombin-induced human platelet aggregation concentration dependently in vitro, with IC(50) values of 7.8 +/- 0.4, 10.1 +/- 0.4 and 18.4 +/- 2.0 micromol/l, respectively. It was also active in inhibiting Ca(2+) ionophore, A23187-induced platelet aggregation, suggesting that J78 may have an inhibitory effect on Ca(2+) mobilization. J78, however, did not alter coagulation parameters such as activated partial thromboplastin time and prothrombin time in human plasma. Taken together, these results suggest that J78 may be a promising antithrombotic agent, and its antithrombotic activity may be due to antiplatelet rather than anticoagulation activity.