Luteoloside protects the vascular endothelium against iron overload injury via the ROS/ADMA/DDAH II/eNOS/NO pathway / 中国天然药物

Iron overload injury is considered to be a part of blood stasis syndrome of arthralgia in traditional Chinese medicine. Its primary therapies include clearing heat and detoxification, activating blood circulation, and removing blood stasis. Lonicera japonica flos (LJF) has long been known as an excellent antipyretic and antidote. Luteoloside (Lut) is one of the main components of LJF and exhibits antioxidant, anti-inflammatory, and cytoprotective properties. However, the protection of Lut against iron overload injury and its underlying mechanisms remain unclear. Therefore, HUVECs were exposed to 50 μmol·L-1 iron dextran for 48 h to establish an iron overload damage model and the effects of Lut were assessed. Our results showed that 20 μmol·L-1 Lut not only increased cell viability and weakened LDH activity, but also significantly up-regulated DDAHⅡ expression and activity, increased p-eNOS/eNOS ratio and NO content, and reduced ADMA content in HUVECs exposed to iron overload. Furthermore, Lut significantly attenuated intracellular/mitochondrial ROS generation, improved SOD, CAT, and GSH-Px activities, reduced MDA content, maintained MMP, inhibited mPTP opening, prevented cyt c from mitochondria released into cytoplasm, reduced cleaved-caspase3 expression, and ultimately decreased cell apoptosis induced by iron overload. The effects of Lut were similar to those of L-arginine (an ADMA competitive substrate), cyclosporin A (a mPTP blocker agent), and edaravone (a free radical scavenger) as positive controls. However, addition of pAD/DDAH II-shRNA adenovirus reversed the above beneficial effects of Lut. In conclusion, Lut can protect HUVECs against iron overload injury via the ROS/ADMA/DDAH II/eNOS/NO pathway. The mitochondria are the target organelles of Lut's protective effects.

Sesamin and vitamin E ameliorate aortic endothelial dysfunction rats induced by D-galactose and aluminum trichloride via regula?tion of endothelial nitric oxide synthase and NAD(P)H oxidase 4 / 中国药理学与毒理学杂志

OBJECTIVE To observe the protective effect of sesamin (Ses) and vitamin E (Vit E) against aortic endothelial dysfunction in rats induced by D-galactose (D-gal) and aluminum trichloride (AlCl3), and explore its conceivable mechanisms. METHODS A model of aortic endothelial dysfunction rats was established by D-gal (180 mg · kg-1, ip) combined with AlCl3 (15 mg · kg-1, ig) for 84 d. Model rats were randomly divided into model, model+Vit E 10 mg·kg-1, model+Ses 160 mg·kg-1, and model+Ses 160 mg · kg-1+Vit E 10 mg · kg-1 groups. After 70 d of treatment with Ses and Vit E, systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean blood pressure (MBP) were measured by tail cuff. The rats were anesthetized by sodium pentobarbital (30 mg·kg-1, ip). Thoracic aortas from the rats were removed and divided into two parts (3 mm in length). The relaxation of the aortic ring induced by acetylcholine (ACh) and sodium nitroprusside was measured. The primary pathologic changes in the aorta were observed by HE staining. Total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2) and nitric oxide (NO) in serum were measured by colorimetric analysis. The expression of endothelial nitric oxide synthase (eNOS) positive cells in the aorta were measured by immunohistochemistry. The expres?sions of eNOS and NAD(P)H oxidase 4 (NOX4) protein in the aortal were detected by Western blotting. RESULTS Compared with the model group, the relaxation response with increase in ACh concentra?tion (1×10-7-1×10-4 mol·L-1) was enhanced (P<0.01) in model+Ses+Vit E, SBP, DBP and MBP decreased (P<0.01), the serum T-AOC and NO level were increased (P<0.01), the serum H2O2 levels were reduced (P<0.01), the eNOS expression was increased (P<0.01) and NOX4 expression was reduced (P<0.01) in each treatment group. Compared with model+Ses, the SBP, DBP and MBP were lower (P<0.01 or P<0.05), the serum H2O2 level was lower (P<0.01), the serum NO level was increased (P<0.05), the eNOS expression level was higher (P<0.01) and the NOX4 expression level was reduced (P<0.05) in model+Ses+Vit E. Compared with the model+Vit E, the serum T-AOC and NO levels were increased (P<0.05), the serum H2O2 level was lower (P<0.01), eNOS expression was increased (P<0.01) and NOX4 expression was reduced (P<0.05) in model+Ses+Vit E group. CONCLUSION Ses and Vit E can ameliorate aortic endothelial dysfunction of rats induced by D-gal and AlCl3 via the regulation of eNOS and NOX4.

Serum Sirtuin 1, 3 and 6 Levels in Acute Myocardial Infarction Patients / Níveis de Sirtuína 1, 3 e 6 em Pacientes com Infarto Agudo do Miocárdio

Abstract Background: Sirtuins may act in many cellular processes like apoptosis, DNA repair and lipid/glucose metabolism. Experimental studies suggested some sirtuin types may have protective effects against endothelial dysfunction, atherosclerosis, cardiac hypertrophy and reperfusion injury. Data about sirtuins in acute myocardial infarction (AMI) patients are scarce. Objectives: To investigate temporal changes of serum sirtuin 1,3 and 6 levels in AMI patients; to compare the serum sirtuin 1,3 and 6 levels between AMI patients and control subjects; and to investigate the association of serum sirtuin 1,3 and 6 levels with prognostic markers of AMI. Methods: Forty patients with AMI and 40 patients with normal coronary arteries were included. Left ventricular ejection fraction (LVEF), serum proBNP, CRP, sirtuin1, sirtuin 3 and sirtuin 6 levels were processed. Peak troponin T levels, GRACE score, first day / second day sirtuin levels were recorded of AMI patients. A p value < 0.05 was considered statistically significant. Results: Serum sirtuin 1,3 and 6 levels in AMI patients were similar to those in normal coronary patients. No temporal change in serum sirtuin 1,3 and 6 levels were found in AMI course. No correlation was evident between the sirtuin levels and the following parameters: proBNP, CRP, peak troponin and LVEF. Baseline sirtuin 1 and 6 levels were positively correlated with reperfusion duration. Baseline sirtuin 3 levels were negatively correlated with GRACE score. Conclusion: Serum sirtuin 1,3 and 6 levels in AMI patients were similar to those in normal coronary patients. This study does not represent evidence of the possible protective effects of sirtuin1, 3 and 6 in AMI patients.

Resumo Fundamento: As sirtuínas podem atuar em muitos processos celulares, como a apoptose, reparo de DNA e metabolismo de lipídios e de glicose. Estudos experimentais sugeriram que alguns tipos de sirtuínas possam ter efeitos protetores contra disfunção endotelial, aterosclerose, hipertrofia cardíaca e lesão decorrente de reperfusão. Dados sobre as sirtuínas em pacientes com infarto agudo do miocárdio (IAM) são escassos. Objetivos: Avaliar as mudanças temporais dos níveis de sirtuína 1, 3 e 6 entre pacientes com IAM e indivíduos controles; investigar a associação entre os níveis de sirtuína 1, 3 e 6 e marcadores prognósticos de IAM. Métodos: Quarenta pacientes com IAM e 40 pacientes com artérias coronárias normais foram incluídos. Foram avaliados fração de ejeção do ventrículo esquerdo (FEVE), concentrações séricas de pró-BNP, proteína C-reativa, sirtuína 1, sirtuína 3 e de sirtuína 6. Pico de troponina T, escore GRACE, concentrações de sirtuínas no primeiro e no segundo dia foram registrados dos pacientes com IAM. Um valor de p<0,05 foi considerado estatisticamente significativo. Resultados: Os níveis de sirtuína 1, 3 e 6 em pacientes com IAM foram similares aos de pacientes com coronária normal. Não foram observadas mudanças temporais nos níveis de sirtuína 1, 3 e 6 no curso do IAM. Nenhuma correlação evidente foi observada dos níveis de sirtuína com os seguintes parâmetros: pró-BNP, proteína C-reativa, pico de troponina e FEVE. Níveis basais de sirtuína 1 e 6 apresentaram correlação positiva com a duração da reperfusão. Os níveis basais de sirtuína 3 correlacionaram-se negativamente com o escore GRACE. Conclusão: Os níveis de sirtuína 1, 3 e 6 em pacientes com IAM foram similares aos de pacientes com artérias coronárias normais. Este estudo não apresenta evidência de possíveis efeitos protetores da sirtuína 1, 3 e 6 em pacientes com IAM.

Does Mean Platelet Volume Decrease in the presence of Coronary Artery Fistula? / O Volume Médio Plaquetário Diminui na Presença de Fístula da Artéria Coronária?

Abstract Background: Coronary artery fistula (CAF) is an abnormal connection that links a coronary artery to a cardiac chamber or another major blood vessel. Several studies have shown the association between mean platelet volume (MPV) and cardiovascular diseases. In the literature, there is no previous study about the association between hematologic parameters and congenital CAF. For this reason, we aimed to investigate the association of MPV with CAF. Methods: 70 patients with normal coronary arteries and 50 with coronary artery fistulas were included. Routine blood and biochemical parameters were measured before the arteriography. Differences between groups for continuous variables were analyzed with t- test or Mann-Whitney test. P values < 0.05 were considered significant. Regression analysis was used to find independent predictors of CAF. Results: Baseline patient demographics, including age and clinical risk factors, were similar between the groups. Compared to the control group, median (IQR) High-density lipoprotein cholesterol (HDL) levels were significantly higher (p=0.04) and MPV levels were significantly lower in the CAF group (8.84 ± 1.71fL vs. 10.43 ± 1.34, p < 0.001). In the multivariate analysis, only MPV was a significant predictor of CAF (p < 0.001, 95% CI for OR: 0.438 (0.306-0.629). A negative correlation was found between MPV and fistulae in Pearson's correlation test (r: -0.454, p < 0.001). An MPV level of < 9,6 fL showed sensitivity, specificity, positive predictive value and negative predictive value of 80%, 68%, 71% and 78% respectively (AUC = 0.766, 95% CI, 0.678-0.854) for the prediction of CAF. Conclusion: The present study suggests that MPV may decrease in patients with CAF.

Resumo Fundamento: A fístula da artéria coronária (FAC) é uma conexão anormal que liga a artéria coronária a uma câmara cardíaca ou outro importante vaso sanguíneo. Vários estudos mostraram a associação entre o volume plaquetário médio (VPM) e as doenças cardiovasculares. Na literatura, não há estudo prévio sobre a associação entre os parâmetros hematológicos e a FAC congênita. Por essa razão, nosso objetivo foi investigar a relação do VPM com a FAC. Métodos: Foram incluídos 70 pacientes com artérias coronárias normais e 50 com fístulas de artérias coronárias. Os parâmetros sanguíneos e bioquímicos de rotina foram medidos antes da arteriografia. As diferenças entre os grupos para as variáveis contínuas foram analisadas com o teste t ou teste de Mann-Whitney. Valores de p < 0,05 foram considerados significativos. A análise de regressão foi utilizada para encontrar preditores independentes de FAC. Resultados: Os dados demográficos basais dos pacientes, incluindo idade e fatores de risco clínicos, foram semelhantes entre os grupos. Comparados à mediana do grupo controle (IIQ), os níveis de HDL-colesterol foram significativamente mais altos (p = 0,04) e os níveis de VPM foram significativamente mais baixos no grupo FAC (8,84 ± 1,71fL vs. 10,43 ± 1,34, p < 0,001). Na análise multivariada, apenas o VPM foi um preditor significativo de FAC (p<0,001, IC 95% para OR: 0,438 (0,306-0,629)). Foi encontrada uma correlação negativa entre o VPM e fístulas no teste de correlação de Pearson (r: -0,454, p < 0,001). Um nível de VPM < 9,6 fL apresentou sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo de 80%, 68%, 71% e 78%, respectivamente (AUC = 0,766, IC 95%, 0,678-0,854) para a previsão de FAC. Conclusão: O presente estudo sugere que o VPM pode diminuir no paciente com FAC.

Serum Uric Acid Levels are Associated with Cardiometabolic Risk Factors in Healthy Young and Middle-Aged Adults / Os Níveis Séricos de Ácido Úrico estão Associados a Fatores de Risco Cardiometabólico em Adultos Jovens e de Meia-Idade Saudáveis

Abstract Background: Observational studies have highlighted an association between serum uric acid (SUA) levels and cardiovascular risk factors. Despite the growing body of evidences, several studies were conducted in older individuals or in carriers of diseases susceptible to affect SUA levels and cardiometabolic risk markers. Objective: To evaluate the relationship of SUA with body adiposity, metabolic profile, oxidative stress, inflammatory biomarkers, blood pressure and endothelial function in healthy young and middle-aged adults. Methods: 149 Brazilian adults aged 20-55 years, both sexes, underwent evaluation of body adiposity, SUA, fasting glucose and insulin, lipid profile, malondialdehyde (MDA), high sensitivity C-reactive protein (hs-CRP), adiponectin, blood pressure and endothelial function. Endothelial function was assessed by the reactive hyperemia index (RHI) derived from peripheral arterial tonometry method. Participants were allocated in two groups according to SUA levels: control group (CG; n = 130; men ≤ 7 mg/dL, women ≤ 6 mg/dL) and hyperuricemia group (HG; n = 19; men > 7 mg/dL, women > 6 mg/dL). A P-value < 0.05 was considered statistically significant. Results: After adjustment for confounders, participants in HG compared with those in CG displayed higher body mass index (BMI): 34.15(33.36-37.19) vs.31.80 (26.26-34.42) kg/m2,p = 0.008, higher MDA: 4.67(4.03-5.30) vs. 3.53(3.10-4.07) ng/mL, p < 0.0001 and lower RHI: 1.68 ± 0.30 vs. 2.05 ± 0.46, p = 0.03). In correlation analysis adjusted for confounders, SUA was positively associated (p < 0.05) with BMI, waist circumference, LDL-cholesterol, triglycerides and MDA, and negatively associated (p < 0.05) with HDL-cholesterol, adiponectin and RHI. Conclusions: This study suggests that in healthy young and middle-aged adults higher SUA levels are associated with higher body adiposity, unfavorable lipid and inflammatory phenotype, higher oxidative stress and impaired endothelial function.

Resumo Fundamento: Estudos observacionais têm destacado uma associação entre níveis de ácido úrico sérico (AUS) e fatores de risco cardiovascular. Apesar do crescente conjunto de evidências, vários estudos foram realizados em indivíduos mais velhos ou em portadores de doenças passíveis de influenciar os níveis de AUS e marcadores de risco cardiometabólico. Objetivo: Avaliar a relação do AUS com adiposidade corporal, perfil metabólico, estresse oxidativo, biomarcadores de inflamação, pressão arterial e função endotelial em adultos jovens e de meia-idade saudáveis. Métodos: 149 adultos, brasileiros, com idades entre 20 e 55 anos, de ambos os sexos, foram submetidos a avaliação de adiposidade corporal, AUS, glicose e insulina de jejum, perfil lipídico, malondialdeído (MDA), proteína C-reativa ultra-sensível (PCR-us), adiponectina, pressão arterial e função endotelial. A função endotelial foi avaliada pelo índice de hiperemia reativa (RHI) derivado do método de tonometria arterial periférica. Os participantes foram divididos em dois grupos de acordo com os níveis de AUS: grupo de controle (GC; n = 130; homens ≤ 7 mg/dL, mulheres ≤ 6mg/dL) e grupo de hiperuricemia (GH; n = 19; homens > 7mg/dL, mulheres > 6mg/dL). Valor de p < 0,05 foi considerado estatisticamente significativo. Resultados: Após ajuste para fatores de confundimento, os participantes do GH comparados aos do GC apresentaram índice de massa corporal (IMC) mais alto: 34,15 (33,36-37,19) vs. 31,80 (26,26-34,42) kg/m2, p = 0,008, MDA mais alto: 4,67(4,03-5,30) vs. 3,53(3,10-4,07) ng/mL, p < 0,0001 e RHI mais baixo: 1,68 ± 0,30 vs. 2,05 ± 0,46, p = 0,03. Na análise de correlação ajustada para fatores de confundimento, o AUS se associou positivamente (p < 0,05) com IMC, circunferência da cintura, LDL colesterol, triglicérides e MDA, e se associou negativamente (p < 0,05) com HDL colesterol, adiponectina e RHI. Conclusões: Este estudo sugere que, em adultos jovens e de meia-idade saudáveis, níveis mais altos de AUS estão associados a maior adiposidade corporal, fenótipo inflamatório e de lipídios desfavorável, maior estresse oxidativo e função endotelial comprometida.

Possible Involvement of "Phosphatidylinositol-3 kinase and endothelial nitric oxide synthase' in experimental obesity induced vascular endothelium dysfunction.

This study was designed to investigate the possible role of Phosphatidylinositol-3 kinase (PI3K) and endothelial nitric oxide synthase (eNOS) in obesity-induced vascular endothelium dysfunction. Wistar rats were fed high fat diet (HFD) for 12 weeks to induce obesity. HFD induce obesity significantly increased parameters employed viz body weight, basal metabolic index (BMI), total fat pad and lipid profile. Furthermore vascular endothelial dysfunction was assessed in terms of decrease in endothelium dependent relaxation and serum nitrate/nitrite level and increase in mean arterial blood pressure. Atorvastatin (30mg/kg,i.p.) was employed in the present study as standard drug to improve vascular endothelial dysfunction. YS-49 (1.6 mg/kg, i.p.) a specific activator of PI3K and atorvastatin in obese rats significantly improves the lipid profile and markedly improved acetylcholine induced endothelium dependent relaxation, serum nitrite/nitrate concentration and mean arterial blood pressure. However, this ameliorative effect of YS-49 has been prevented by L-NAME (25 mg/kg, i.p.), an inhibitor of eNOS. Further wortmannin (100μg/kg, i.p.),a specific PI3K inhibitor improves the anthropometric parameters and lipid profile but did not show any significant effect on acetylcholine dependent relaxation, serum nitrate/nitrite level and mean arterial blood pressure. Therefore, it may be concluded that PI3K and eNOS pathway is dysregulated in obesity induced vascular endothelial dysfunction and YS-49, a PI3K activator improves vascular endothelial function in eNOS and nitric oxide dependent manner. Abbreviations (Ach): Acetylcholine, (Ang-II): Angiotensin II, (BMI): Body Mass Index, (Enos): Endothelial Nitric Oxide, Synthase, (HO-1): Heme Oxygenase, (Kcl): Pottesium Cloride, (MAP): Mean Arterial Pressure, (NO): Nitric Oxide, (PI3K): Phosphatidylinositol 3-Kinase, (SNP ): Sodium Nitroprusside, (VED): Vascular Endothelium Dysfunction

Ameliorative effect of daidzein: A caveolin-1 inhibitor in vascular endothelium dysfunction induced by ovariectomy.

Estrogen deficiency was produced in female Sprague-Dawley rats by surgical removal of both the ovaries and these animals were used 4 weeks later. Endothelium-dependent and endothelium-independent relaxations due to acetylcholine and sodium nitroprusside were observed respectively, in isolated rat thoracic aortic ring preparation. Extent of lipid peroxidation was measured by estimating serum TBARS. Integrity of vascular endothelium was assessed using hematoxylin and eosin staining. Generation of nitric oxide was measured indirectly, by estimating serum and urinary nitrite/nitrate concentration. Ovariectomy produced significant vascular endothelial dysfunction, measured in terms of reduced acetylcholine-induced endothelium-dependent vasorelaxation, serum and urinary nitrite/nitrate concentration and impairment of integrity of vascular endothelium. Administration of daidzein (0.2 mgkg-1day-1, sc 0.4 mgkg-1day-1, sc and 0.8 mgkg-1day-1, sc) and Atorvastatin (30 mgkg-1day-1, po Positive Control) for one week markedly improved vascular endothelial dysfunction due to increase in nitric oxide bioavailability perhaps by inhibiting caveolin-1 and activation of PI3K-AKT pathway.

VEGF Deficit is Involved in Endothelium Dysfunction in Preeclampsia / 华中科技大学学报（医学）（英德文版）

This study examined the association of expression of vascular endothelial growth factor(VEGF),a promoter of angiogenesis,with endothelium dysfunction in preeclampsia.The level of VEGF protein and mRNA in the placenta and peripheral blood samples of 30 preeclampsia patients and 30 normotensive pregnant women was measured by immunohistochemistry,real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA),respectively.VEGF expression in the human umbilical vein endothelial cells(HUVECs)was blocked by small interfering RNAs(siRNAs).The monolayer barrier function of HUVECs was determined by measuring the fluorescence intensity of BSA that crossed the HUVEC monolayers.The cell proliferation and cell-secreted nitric oxide(NO)level were detected by MTT method and nitrate reductase assay,respectively.The results showed that VEGF was expressed in the syncytiotrophoblasts and endothelial cells of vessels and capillaries in the placenta tissue.The serum level of VEGF in the preeclampsia patients was significantly decreased as compared with that in normal pregnant subjects,although VEGF mRNA expression in the placenta tissue of preeclampsia patients remained still high.Moreover,VEGF deficit could lead to endothelium cell dysfunction,and the administration of VEGF could protect endothelium cells from injury.It was concluded that lack of VEGF contributes to endothelium dysfunction,which may lead to the occurrence and development of preeclampsia.

Device for Endothelium Dysfunction Detection Based on Photoplethysmographic Method / 医疗卫生装备

Objective To evaluate endothelium dysfunction of arteries for early preventing atherosis. Methods A new portable device for detection of endothelium dysfunction by photoplethysmographic method was designed, which depended on the principle of reactive hyperaemia. Results Experiments between hypertensive and normotensive showed that different degree endothelium dysfunction had emerged among hypertensive. Conclusion The proposed device has potential benefit to research, diagnosis and therapy evaluation for subclinic arterial disease.