ABSTRACT
Objective: To investigate the effect of Salvianolic acid B (Sal B) on the apoptosis of human umbilical vein endothelial cells (HUVECs) induced by intermittent high glucose and to explore the possible mechanisms. Methods: HUVECs were preincubated with Sal B for 24 h, followed by incubation with intermittent high glucose (IHG, 5.5 mmol/L 12 h, 33.3 mmol/L 12 h) for 72 h. The viability of the HUVECs was determined by MTT assay, and the cells apoptosis was measured flow cytometry, respectively. The levels of nitric oxide (NO), total antioxidant capacity (T-AOC), malondialdehyde (MDA), and Caspase-3 activity were determined by colorimetric method. Intracellular ROS was evaluated by fluorescent microscopy. The protein levels of NOX4, p-eNOS, BAX, and BCL-2 were determined by Western-blot. Results: Pretreatment with Sal B significantly ameliorated IHG-induced cells injury as was manifested by increased cell viability, up-regulated eNOS activation, and promoted the release of NO in HUVECs (P < 0.05 or P < 0.01). Sal B evidently suppressed IHG-induced cell apoptosis, down-regulated the expression of BAX protein and up-regulated the expression of BCL-2 protein. The activity of Capase-3 was also significantly reduced. Pre-incubation with Sal B led to a significant enhancement of antioxidant capacity and a reduction of NOX4 protein expression, accompanied by a remarkable decrease of intracellular ROS and MDA content (P < 0.05 or P < 0.01). Conclusion: Sal B is capable of suppressing IHG-induced injury and apoptosis in HUVECs, which might be attributed to the attenuation of oxidative stress, regulation of BCL-2/BAX protein expression, and subsequent suppression of Caspase-3 activity.
ABSTRACT
OBJECTIVE: To observe the effect of sesamin (Ses) on pulmonary vascular remodeling in rats with monocrotaline ( MCT)-induced pulmonary hypertension (PH). METHOD: Totally 48 male Sprague-Dawley (SD) rats were fed adaptively for one week and then divided into the normal control group, the MCT group, the MCT +Ses (50 mg x kg(-1)) group and the MCT + Ses (100 mg x kg(-1)) group, with 12 rats in each group. The PH rat model was induced through the subcutaneous injection with MCT(60 mg x kg(-1)). After the administration for four weeks, efforts were made to measure the right ventricular systolic pressure( RVSP) and mean pulmonary artery pressure (mPAP) through right jugular vein catheterization, and isolate right ventricle( RV) and left ventricle( LV) +septum (S) and measure their length to calculate RV/ ( LV + S) and ratio of RV to tibial length. Pathologic changes in arterioles were observed by HE staining. Masson's trichrome stain was used to demonstrate changes in collagen deposition of arterioles. The alpha-smooth muscle actin (alpha-SMA) expression in pulmonary arteries was measured by immunohistochemisty. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) content in pulmonary arteries were determined by the colorimetric method. The protein expressions of collagen I, NOX2 and NOX4 were analyzed by Real-time PCR and Western blot. RESULT: After the administration for 4 weeks, Ses could attenuate RVSP and mPAP induced by MCT, RV/ (LV + S) and ratio of RV to Tibial length, alpha-SMA and collagen I expressions and remodeling of pulmonary vessels and right ventricle. Meanwhile, Ses could obviously inhibit the expressions of NOX2, NOX4 and MDA content and increase T-AOC. CONCLUSION: Sesamin could ameliorate pulmonary vascular remodeling induced by monocrotaline in PH rats. Its mechanism may be related to expressions of NOX2 and NOX4 expression and reduction in oxidative stress injury.
Subject(s)
Dioxoles/administration & dosage , Drugs, Chinese Herbal/administration & dosage , Hypertension, Pulmonary/drug therapy , Lignans/administration & dosage , Vascular Remodeling/drug effects , Animals , Disease Models, Animal , Humans , Hypertension, Pulmonary/enzymology , Hypertension, Pulmonary/genetics , Hypertension, Pulmonary/physiopathology , Lung/blood supply , Lung/enzymology , Lung/metabolism , Male , Membrane Glycoproteins/genetics , Membrane Glycoproteins/metabolism , Monocrotaline/adverse effects , NADPH Oxidase 2 , NADPH Oxidase 4 , NADPH Oxidases/genetics , NADPH Oxidases/metabolism , Pulmonary Artery/drug effects , Pulmonary Artery/metabolism , Pulmonary Artery/physiopathology , Rats , Rats, Sprague-DawleyABSTRACT
Advanced glycation end products (AGEs), the direct modulators of ß-cells, have been shown to cause insulin-producing ß-cell dysfunction and apoptosis through increase of intracellular reactive oxygen species (ROS) production. Sesamin has been demonstrated to possess antioxidative activity. This study was designed to investigate whether sesamin protects against AGEs-evoked ß-cell damage via its antioxidant property. The effects of sesamin were examined in C57BL/6J mice and MIN6 cell line. In in vivo studies, mice were intraperitoneally injected with AGEs (120 mg/kg) and orally treated with sesamin (160 mg/kg) for four weeks. Intraperitoneal glucose tolerance and insulin releasing tests were performed. Insulin content, ROS generation and ß-cell apoptosis in pancreatic islets were also measured. In in vitro studies, MIN6 cells were pretreated with sesamin (50 or 100 µM) and then exposed to AGEs (200 mg/L) for 24 h. Insulin secretion, ß-cell death, ROS production as well as expression and activity of NADPH oxidase were determined. Sesamin treatment obviously ameliorated AGE-induced ß-cell dysfunction and apoptosis both in vivo and in vitro. These effects were associated with decreased ROS production, down-regulated expression of p67(phox) and p22(phox), and reduced NADPH oxidase activity. These results suggest that sesamin protects ß-cells from damage caused by AGEs through suppressing NADPH oxidase-mediated oxidative stress.
Subject(s)
Antioxidants/pharmacology , Apoptosis/drug effects , Dioxoles/pharmacology , Glycation End Products, Advanced/metabolism , Insulin-Secreting Cells/drug effects , Lignans/pharmacology , Animals , Cells, Cultured , Insulin/metabolism , Insulin Secretion , Insulin-Secreting Cells/metabolism , Mice , Mice, Inbred C57BL , NADPH Oxidases/antagonists & inhibitors , NADPH Oxidases/metabolism , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolismABSTRACT
BACKGROUND: The blood pressure lowering effect of sesamin has been demonstrated to be associated with the increase in vascular nitric oxide (NO) biological activity by our previous studies and others. The present study was designed to explore the underlying mechanisms involved in the effect of sesamin on aortic NO bioactivity in spontaneously hypertensive rats (SHRs). METHODS: Sesamin was orally administered for 8 consecutive weeks in SHRs. Systolic blood pressure (SBP) was measured using the tail-cuff method. The aortas were isolated and in vitro vascular reactivity studies were performed. Superoxide anion production in carotid arteries was assessed by dihydroethidium fluorescence staining. The protein expression of endothelial nitric oxide synthase (eNOS), phosphorylated eNOS (P-eNOS), dihydrofolate reductase (DHFR), nicotinamide adenine dinucleotide phosphate oxidase subunit p47phox, and copper, zinc superoxide dismutase (Cu/Zn-SOD) in aortas was detected by Western blotting. The dimeric form of eNOS in aortas was determined by low-temperature sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Aortic level of nitrotyrosine and activities of antioxidant enzymes, namely, total SOD (T-SOD), glutathione peroxidase (GPx) and catalase were also detected. RESULTS: In SHRs, sesamin treatment reduced SBP, improved vascular relaxation induced by acetylcholine and enhanced aortic NO bioactivity. Sesamin treatment enhanced NO biosynthesis in SHR aortas was due to upregulated P-eNOS and suppressed eNOS uncoupling, and the latter effect might be attributed to decreased nitrotyrosine and upregulated DHFR. Sesamin also reduced the NO oxidative inactivation and decreased the superoxide anion production through downregulation of p47(phox) and amelioration of eNOS uncoupling. In addition, sesamin treatment did not alter the levels of GPx and catalase activity but obviously reduced the compensatory elevated T-SOD activity and Cu/Zn-SOD protein expression. CONCLUSION: Chronic treatment with sesamin could reduce hypertension and improve endothelial dysfunction through enhancement of NO bioactivity in SHR aortas.
Subject(s)
Antihypertensive Agents/pharmacology , Aorta/drug effects , Dioxoles/pharmacology , Lignans/pharmacology , Nitric Oxide/metabolism , Animals , Antioxidants/metabolism , Aorta/metabolism , Blood Pressure/drug effects , Blotting, Western , Down-Regulation/drug effects , Electrophoresis, Polyacrylamide Gel , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Hypertension/drug therapy , Hypertension/physiopathology , Male , Nitric Oxide Synthase Type III/metabolism , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Superoxide Dismutase/metabolism , Up-Regulation/drug effectsABSTRACT
It has also been shown that the decreased expression of eukaryotic translation initiation factor 3a (eIF3a) by L-mimosine caused cell cycle arrest. Our previous study has found that eIF3a is involved in bleomycin-induced pulmonary fibrosis. Whether the eIF3a/p27 signal pathway is involved in the inhibitory effect of L-mimosine on bleomycin-induced pulmonary fibrosis remains unknown. Pulmonary fibrosis was induced by intratracheal instillation of bleomycin (5 mg/kg) in rats. Primary pulmonary fibroblasts were cultured to investigate the proliferation by BrdU incorporation method and flow cytometry. The expression of eIF3a, p27, α-SMA, collagen I and collagen III was analyzed by qPCR and Western blot. In vivo, L-mimosine treatment significantly ameliorated the bleomycin-mediated histological fibrosis alterations and blocked collagen deposition concomitantly with reversing bleomycin-induced expression up-regulation of eIF3a, α-SMA, collagen I and collagen III (both mRNA and protein) and expression down- regulation of p27. In vitro, L-mimosine remarkably attenuated proliferation of pulmonary fibroblasts and expression of α-SMA, collagen I and collagen III induced by TGF-ß1, and this inhibitory effect of L-mimosine was accompanied by inhibiting eIF3a expression and increasing p27 expression. Knockdown of eIF3a gene expression reversed TGF-ß1-induced proliferation of fibroblasts, down-regulation of p27 expression and up-regulation of α-SMA, collagen I, and collagen III expression. These results suggest that L-mimosine inhibited the progression of bleomycin-induced pulmonary fibrosis in rats via the eIF3a/p27 pathway.
Subject(s)
Eukaryotic Initiation Factor-3/metabolism , Fibroblasts/drug effects , Lung/pathology , Mimosine/administration & dosage , Pulmonary Fibrosis/drug therapy , Actins/genetics , Actins/metabolism , Animals , Bleomycin/administration & dosage , Cell Proliferation/drug effects , Cells, Cultured , Collagen/metabolism , Cyclin-Dependent Kinase Inhibitor p27/genetics , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Eukaryotic Initiation Factor-3/genetics , Fibroblasts/physiology , Gene Expression Regulation/drug effects , Humans , Male , Mimosine/adverse effects , Pulmonary Fibrosis/chemically induced , RNA, Small Interfering/genetics , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , Transforming Growth Factor beta/metabolismABSTRACT
Crocetin, a naturally occurring carotenoid, possesses antioxidant and antiatherosclerotic properties, of which the underlying mechanism remains unclear. In the present study, we examined the effects of crocetin (0.1, 1, 10 µmol·L(-1)) on angiotensin II (Ang II, 0.1 µmol·L(-1)) induced expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and monocyte-endothelial cell adhesion. The effects of crocetin on the activation of nuclear factor kappa B (NF-κB) and intracellular reactive oxygen species (ROS) were also observed. The results demonstrated that crocetin notably suppressed Ang II induced NF-κB activation (P<0.01) and VCAM-1 expression (P<0.05, P<0.01) in HUVECs, accompanied by a markedly reduced monocyte-endothelial cell adhesion (P<0.05, P<0.01). In addition, preincubation with crocetin resulted in a significant enhancement of cellular antioxidant capacity (P<0.05, P<0.01), while Ang II induced intracellular ROS decreased markedly (P<0.05, P<0.01). These results indicated that crocetin was capable of suppressing Ang II induced VCAM-1 expression and monocyte-endothelial cell adhesion by suppression of NF-κB activation, which might be derived from the enhancement of antioxidant capacity and subsequent reduction of intracellular ROS.
Subject(s)
Carotenoids/pharmacology , Cell Adhesion/drug effects , Human Umbilical Vein Endothelial Cells/drug effects , Monocytes/cytology , Vascular Cell Adhesion Molecule-1/metabolism , Angiotensin II/metabolism , Antioxidants/pharmacology , Human Umbilical Vein Endothelial Cells/cytology , Human Umbilical Vein Endothelial Cells/metabolism , Humans , NF-kappa B/metabolism , Reactive Oxygen Species/metabolism , Vitamin A/analogs & derivativesABSTRACT
BACKGROUND: Chrysin (5,7-dihydroxyflavone) inhibits platelet-derived growth factor-induced vascular smooth muscle cell proliferation and arterial intima hyperplasia. This study aims to investigate the effects of chrysin on rat pulmonary vascular remodeling in hypoxia-induced pulmonary hypertension (PH). METHODS: Sprague-Dawley rats were continuously exposed to 10% O2 for 4 weeks to induce PH. The effect of chrysin (50 or 100 mg/kg/d, subcutaneous) on vascular remodeling was investigated in hypoxia-induced PH model. At the end of the experiments, the indexes for pulmonary vascular remodeling and right ventricle hypertrophy were measured by vascular medial wall thickness and the ratio of right ventricle to (left ventricle plus septum). The expressions of NOX4, collagen I, and collagen III were analyzed by immunohistochemistry, real-time PCR, or western blotting. The proliferation of cultured pulmonary artery smooth muscle cells (PASMCs) was determined by BrdU incorporation and flow cytometry. The levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were also determined by thiobarbituric acid reactive substances assay and 2'7'-dichlorofluorescein diacetate method. RESULTS: Chrysin treatment for 4 weeks significantly attenuated pulmonary vascular remodeling and improved collagen accumulation and down-regulated collagen I and collagen III expressions, accompanied by downregulation of NOX4 expression in the pulmonary artery (P = 0.012 for 50 mg/kg/d, P < 0.001 for 100 mg/kg/d) and lung tissue (P = 0.026, P < 0.001). In vitro, chrysin (1, 10, and 100 µM) remarkably attenuated PASMC proliferation (P = 0.021 for 1 µM, P < 0.001 for 10 µM, and P < 0.001 for 100 µM), collagen I expression (P = 0.035, P < 0.001, and P < 0.001), and collagen III expression (P = 0.027, P < 0.001, and P < 0.001) induced by hypoxia, and these inhibitory effects of chrysin were accompanied by inhibition of NOX4 expression (P = 0.019, P < 0.001, and P < 0.001), ROS production (P = 0.038, P < 0.001, and P < 0.001), and MDA generation (P = 0.024, P < 0.001, and P < 0.001). CONCLUSIONS: This study demonstrated that chrysin treatment in hypoxia-induced PH in rats reversed the hypoxia-induced (1) elevations of NOX4 expression, (2) productions of ROS and MDA, (3) proliferation of PASMC, and (4) accumulation of collagen.
ABSTRACT
OBJECTIVE: To investigate the effect of serum containing sesamin on angiotension II (Ang II)-induced apoptosis in rat cardiomyocytes and the possible mechanisms. METHODS: H9c2 rat cardiomyocytes were preincubated with serum containing sesamin or blank serum for 12 h, followed by incubation with Ang II for 24 h. Cell viability was assessed by MTT assay and cell apoptosis was evaluated by flow cytometric analysis. Protein expression of BCL-2, BAX, Caspase-3, p47phox and superoxide dismutase (SOD) was determined by Western blot analysis. Levels of intracellular reactive oxygen species (ROS), total antioxidant capacity (T-AOC) and malondialdehyde (MDA) were measured colorimetrically. RESULTS: Preincubation with serum containing sesamin significantly improved cell viability and suppressed cell apoptosis in H9c2 rat cardiomyocytes exposed to Ang II (P < 0.05 or P < 0.01), with the expression of BAX, Caspase-3 and p47phox protein down-regulated and BCL-2 and SOD protein up-regulated markedly (P < 0.05 or P < 0.01). The levels of T-AOC were effectively increased in serum containing sesamin groups, while the levels of intracellular ROS and MDA contents were decreased significantly (P < 0.05 or P < 0.01). Control serum had no influence on the above mentioned measurements. CONCLUSION: Sesamin is capable of suppressing Ang II-induced apoptosis in H9c2 rat cardiomyocytes, which might be derived, at least partly, from amelioration of oxidative stress, regulation of BAX/BCL-2 protein expression and suppression of Caspase-3 protein expression.
Subject(s)
Angiotensin II/adverse effects , Apoptosis/drug effects , Dioxoles/pharmacology , Lignans/pharmacology , Myocytes, Cardiac/drug effects , Animals , Antioxidants/metabolism , Caspase 3/metabolism , Cell Line , Cell Survival , Malondialdehyde/metabolism , Oxidative Stress , Proto-Oncogene Proteins c-bcl-2/metabolism , Rats , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolism , bcl-2-Associated X Protein/metabolismABSTRACT
The aim of the present study is to investigate the protective effect of chrysin (5,7-dihydroxyflavone) on right ventricular remodeling in a rat model of monocrotaline-induced pulmonary arterial hypertension (PAH). PAH rats were induced by a single injection of monocrotaline (60 mg x kg(-1), sc) and were administered with chrysin (50 or 100 mg x kg(-1) x d(-1)) for 4 weeks. At the end of experiment, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored via the right jugular vein catheterization into the right ventricle. Right ventricle (RV) to left ventricle (LV) + septum (S) and RV to tibial length were calculated. Right ventricular morphological change was observed by HE staining. Masson's trichrome stain was used to demonstrate collagen deposition. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) levels in right ventricle were determined according to the manufacturer's instructions. The expressions of collagen I, collagen III, NADPH oxidase 4 (NOX4) and nuclear factor-kappa B (NF-κB) were analyzed by immunohistochemisty, qPCR and (or) Western blot. The results showed that chrysin treatment for 4 weeks attenuated RVSP, mPAP and right ventricular remodeling index (RV/LV+S and RV/Tibial length) of PAH rats induced by monocrotaline. Furthermore, monocrotaline-induced right ventricular collagen accumulation and collagen I and collagen III expression were both significantly suppressed by chrysin. The expressions of NOX4, NF-κB and MDA contents were obviously decreased, while the T-AOC was significantly increased in right ventricule from PAH rats with chrysin treatment. These results suggest that chrysin ameliorates right ventricular remodeling of PAH induced by monocrotaline in rats through its down-regulating of NOX4 expression and antioxidant activity, and inhibiting NF-κB expression and collagen accumulation.
Subject(s)
Flavonoids/pharmacology , Heart Ventricles/drug effects , Hypertension, Pulmonary/metabolism , NADPH Oxidases/metabolism , NF-kappa B/metabolism , Animals , Blotting, Western , Collagen/metabolism , Disease Models, Animal , Heart Ventricles/metabolism , Hypertension, Pulmonary/chemically induced , Monocrotaline/toxicity , NADPH Oxidase 4 , Rats , Ventricular Remodeling/drug effectsABSTRACT
BACKGROUND: Recent studies have demonstrated that side population (SP) cells isolated from various cancer cell lines and primary tumors possess stem cell-like properties. Sesamin, a food-derived agent, possesses anti-cancer activities both in vitro and in vivo. The present study was designed to determine whether sesamin also have effects on cancer stem-like SP cells from gallbladder cancer (GBC). METHODS: In this study, we sorted SP cells by flow cytometry. SP cells were cultured and treated with sesamin. Tumor-sphere formation, colony formation, Matrigel invasion and tumorigenic potential were determined. Expression of nuclear NF-κB, IL-6, p-Stat3, Twist, E-cadherin and Vimentin was measured by Western blot, immunofluorescence staining or RT-PCR analysis. Nuclear NF-κB activity and IL-6 protein level were assessed with ELISA. Xenograft tumors were generated in nude mice. RESULTS: After treated with sesamin, SP cells differentiated into cells expressing the epithelial marker (E-cadherin). Sesamin effectively affected SP cells stem cell-like characteristics (i.e., tumor-sphere formation, colony-formation, Matrigel invasion), weakened the drug-resistance of SP cells and inhibited tumor growth both in vitro and in vivo. Treatment with sesamin significantly reduced the expression of nuclear NF-κB, IL-6, p-Stat3, Twist and Vimentin (a mesenchymal marker) in SP cells. Nuclear NF-κB activity and IL-6 level were also decreased after treatment with sesamin. CONCLUSION: Food-derived sesamin directs the epithelial differentiation of cancer stem-like SP cells from GBC, which is associated with attenuation of NF-κB-IL-6-Stat3-Twist signal pathway.
Subject(s)
Dioxoles/pharmacology , Gallbladder Neoplasms/drug therapy , Gallbladder Neoplasms/pathology , Lignans/pharmacology , Neoplastic Stem Cells/drug effects , Side-Population Cells/drug effects , Analysis of Variance , Animals , Cadherins/metabolism , Carcinoma/drug therapy , Carcinoma/metabolism , Carcinoma/pathology , Cell Differentiation/drug effects , Cell Line, Tumor , Female , Gallbladder Neoplasms/metabolism , Humans , Interleukin-6/metabolism , Mice , Mice, Nude , NF-kappa B/metabolism , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Side-Population Cells/metabolism , Side-Population Cells/pathology , Signal Transduction/drug effects , Xenograft Model Antitumor AssaysABSTRACT
OBJECTIVE: To investigate the protective effect and mechanism of sequoyitol (Sep) on high glucose-induced human umbilical vein endothelial cells (HUVECs) injury. METHODS: HUVECs were cultured with high glucose (30 mmol/L) in the presence or absence of sequoyitol (0.1, 1 and 10 micromol/L) for 24 h. Cell proliferation was measured by BrdU marking and cell cycle was detected by flow cytometry. 2', 7'-dichlorofluorescein diacetate was used to evaluate intracellular reactive oxygen species (ROS) levels. The NO, malonydialdehyde (MDA) and H2O2 levels were determined by colorimetric method according to the manufacturer's instructions. The expression of endothelial nitric oxide synthase (eNOS) and NADPH oxidase 4 (NOX4) were measured by real-time PCR and Western blot. RESULTS: In the present study, we found that sequoyitol pretreatment for 1 h significantly decreased cell injury, promoted cell proliferation. Meanwhile sequoyitol significantly down-regulated NOX4 expression and decreased the level of ROS, MDA and H2O2 and obviously increased NO levels and up-regulated eNOS expression. CONCLUSION: Sequoyitol alleviates high glucose-induced cell injuries in HUVECs via inhibiting oxidative stress and up-regulating eNOS expression.
Subject(s)
Human Umbilical Vein Endothelial Cells/metabolism , Inositol/analogs & derivatives , NADPH Oxidases/metabolism , Nitric Oxide Synthase Type III/metabolism , Cell Proliferation , Cells, Cultured , Glucose/toxicity , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Hydrogen Peroxide/metabolism , Inositol/pharmacology , Malondialdehyde/metabolism , NADPH Oxidase 4 , Oxidative Stress , Reactive Oxygen Species/metabolismABSTRACT
The aim of the present study is to investigate the effects of sequoyitol (Seq) on expression of eNOS and NOX4 in aortas of type 2 diabetic rats. Type 2 diabetic rats induced by high fat and high sugar diet and low dose of streptozotocin (STZ, 35 mg x kg(-1)) and were administered Seq (12.5, 25 and 50 mg x kg(-1) x d(-1)) for 6 weeks. The fasting blood glucose (FBG) and body weight were tested. Acetylcholine (Ach) induced endothelium-dependent relaxation and sodium nitroprusside (SNP) induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. Aortic morphological change was observed with HE staining. The level of serum insulin was measured by radioimmunoassay. The total antioxidative capacity (T-AOC), malondialdehyde (MDA) and NO levels in aortas were determined according to the manufacturer's instructions. In addition, the expressions of eNOS and NOX4 in aortas were measured by immunohistochemisty, real-time PCR or Western blotting. The results showed that Seq significantly decreased FBG and insulin resistance, and improved aortic endothelium-dependent vasorelaxation function. The expressions of NOX4 and MDA content were obviously decreased, while the expression of eNOS, the levels of NO and T-AOC increased significantly in aortas of diabetic rats with Seq treatment. In conclusion, Seq protects against aortic endothelial dysfunction of type 2 diabetic rats through down-regulating expression of NOX4 and up-regulating eNOS expression.
Subject(s)
Aorta/metabolism , Diabetes Mellitus, Experimental/metabolism , Diabetes Mellitus, Type 2/metabolism , Inositol/analogs & derivatives , NADPH Oxidases/metabolism , Nitric Oxide Synthase Type III/metabolism , Animals , Aorta/pathology , Blood Glucose/metabolism , Body Weight , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/physiopathology , Diabetes Mellitus, Type 2/chemically induced , Diabetes Mellitus, Type 2/physiopathology , Hypoglycemic Agents/pharmacology , Inositol/pharmacology , Insulin/blood , Insulin Resistance , Male , Malondialdehyde/metabolism , NADPH Oxidase 4 , Nitric Oxide/metabolism , Oxidation-Reduction/drug effects , Rats , Rats, Sprague-Dawley , Streptozocin , Vasodilation/drug effectsABSTRACT
Sequoyitol decreases blood glucose, improves glucose intolerance, and enhances insulin signaling in ob/ob mice. The aim of this study was to investigate the effects of sequoyitol on diabetic nephropathy in rats with type 2 diabetes mellitus and the mechanism of action. Diabetic rats, induced with a high-fat diet and a low dose of streptozotocin, and were administered sequoyitol (12.5, 25.0, and 50.0 mg·(kg body mass)(-1)·d(-1)) for 6 weeks. The levels of fasting blood glucose (FBG), serum insulin, blood urea nitrogen (BUN), and serum creatinine (SCr) were measured. The expression levels of p22(phox), p47(phox), NF-κB, and TGF-ß1 were measured using immunohistochemisty, real-time PCR, and (or) Western blot. The total antioxidative capacity (T-AOC), as well as the levels of malondialdehyde (MDA) and reactive oxygen species (ROS) were also determined. The results showed that sequoyitol significantly decreased FBG, BUN, and SCr levels, and increased the insulin levels in diabetic rats. The level of T-AOC was significantly increased, while ROS and MDA levels and the expression of p22(phox), p47(phox), NF-κB, and TGF-ß1 were decreased with sequoyitol treatment both in vivo and in vitro. These results suggested that sequoyitol ameliorates the progression of diabetic nephropathy in rats, as induced by a high-fat diet and a low dose of streptozotocin, through its glucose-lowering effects, antioxidant activity, and regulation of TGF-ß1 expression.
Subject(s)
Diabetes Mellitus, Experimental/drug therapy , Diabetic Nephropathies/drug therapy , Diet, High-Fat , Hypoglycemic Agents/therapeutic use , Inositol/analogs & derivatives , Animals , Cell Survival/drug effects , Diabetes Mellitus, Experimental/etiology , Diabetes Mellitus, Experimental/metabolism , Diabetic Nephropathies/etiology , Diabetic Nephropathies/metabolism , Hypoglycemic Agents/pharmacology , Inositol/pharmacology , Inositol/therapeutic use , Male , Rats , Rats, Sprague-Dawley , Reactive Oxygen Species/metabolism , StreptozocinABSTRACT
OBJECTIVE: To investigate the protective effects of rutaecarpine (Rut) on right ventricular remodeling in rats with monocrotaline-induced pulmonary hypertension (PH). METHOD: Forty-eight SD rats were fed adaptively for 1 week and then were randomly divided into the following 4 groups (n = 12): normal control group, monocrotaline (MCT) treatment group, MCT treatment with Rut (20 mg/kg)group and MCT treatment with Rut (40 mg/kg) group. PH rats were induced by a single injection of monocrotaline (60 mg/kg, sc) and were administered with Rut (20 or 40 mg/kg/d) for 4 weeks. At the end of experiment, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored via the right jugular vein catheterization into the right ventricle. The ratio of right ventricle (RV) to left ventricle (LV) + septum (S) and the ratio of RV to tibial length were calculated. Right ventricular morphological changes were deserved by HE staining. Masson's trichrome staining was used to display collagen deposition. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) levels in right ventricle were determined according to the manufacturer's instructions. mRNA and protein expression levels of NOX4, collagen I and collagen III were analyzed by immunohistochemisty, real-time PCR and Western blot. RESULTS: The results showed that Rut treatment for 4 weeks attenuated RVSP, mPAP and right ventricular remodeling index (RV/LV + S and RV/Tibial length) of PH rats induced by monocrotaline. Furthermore, the right ventricular collagen deposition and collagen I and collagen I expression induced by MCT were both significantly suppressed by Rut. The expression levels of NOX4 and MDA were obviously decreased, while the T-AOC was significantly increased in right ventricular from PH rats treated with Rut. CONCLUSION: These results suggested that Rut ameliorates the right ventricular remodeling in rats with PH induced by MCT through down-regulating of NOX4 expression and collagen accumulation.
Subject(s)
Hypertension, Pulmonary/drug therapy , Indole Alkaloids/pharmacology , Quinazolines/pharmacology , Ventricular Remodeling/drug effects , Animals , Antioxidants/metabolism , Heart Ventricles/metabolism , Hypertension, Pulmonary/chemically induced , Male , Malondialdehyde/metabolism , Monocrotaline/adverse effects , NADPH Oxidase 4 , NADPH Oxidases/metabolism , RatsABSTRACT
This study is to observe the effects of sequoyitol on the expression of NADPH oxidase subunits p22 phox and p47 phox in rats with type 2 diabetic liver diseases. The model of high fat and high sugar diet as well as intraperitoneal injection of small dose of streptozotocin (STZ, 35 mg x kg(-1)) induced diabetic rat liver disease was used. After sequoyitol (50, 25 and 12.5 mg x kg(-1)) was administrated for 6 weeks, the contents of blood glucose (BG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), NO and insulin (Ins) were measured, liver p22 phox and p47 phox mRNA content was determined with real-time PCR and the expression of p22 phox and p47 phox protein was examined by Western blotting. In addition, pathological changes in liver were observed with HE staining. Sequoyitol could reduce the content of fasting blood glucose, ALT, AST, Ins and H2O2, restore insulin sensitive index (ISI) and weight, elevate liver tissue T-AOC and NO content, reduce the NADPH oxidase subunit liver tissue p22 phox and p47 phox mRNA and protein expression, as well as ameliorate liver pathologic lesions. The results showed that sequoyitol can ease the type 2 diabetic rat liver oxidative stress by lowering NADPH oxidase expression.
Subject(s)
Diabetes Mellitus, Experimental/metabolism , Inositol/analogs & derivatives , Liver Diseases/metabolism , NADPH Oxidases/metabolism , Alanine Transaminase/blood , Animals , Aspartate Aminotransferases/blood , Blood Glucose/metabolism , Diabetes Mellitus, Experimental/chemically induced , Hydrogen Peroxide/metabolism , Hypoglycemic Agents/pharmacology , Inositol/pharmacology , Insulin/blood , Liver/metabolism , Liver/pathology , Male , NADPH Oxidases/genetics , Nitric Oxide/metabolism , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , RNA, Messenger/metabolism , Random Allocation , Rats , Rats, Sprague-Dawley , StreptozocinABSTRACT
AIM: Sesamin is one of the major lignans in sesame seeds with antihyperlipidemic, antioxidative and antihypertensive activities. The aim of this study was to examine the effects of sesamin on arterial function in spontaneously hypertensive rats (SHRs). METHODS: SHRs were orally administered sesamin (40, 80 and 160 mg·kg(-1)·d(-1)) for 16 weeks. After the rats were killed, thoracic aortas were dissected out. The vasorelaxation responses of aortic rings to ACh and nitroprusside were measured. The expression of eNOS and NADPH oxidase subunits p47(phox) and p22(phox) in aortas were detected using Western blotting and immunohistochemistry. Aortic nitrotyrosine was measured with ELISA. The total antioxidant capacity (T-AOC) and MDA levels in aortas were also determined. RESULTS: The aortic rings of SHRs showed significantly smaller ACh-induced and nitroprusside-induced relaxation than those of control rats. Treatment of SHRs with sesamin increased both the endothelium-dependent and endothelium-independent relaxation of aortic rings in a dose-dependent manner. In aortas of SHRs, the level of T-AOC and the expression of nitrotyrosine, p22(phox) and p47(phox) proteins were markedly increased, while the level of MDA and the expression of eNOS protein were significantly decreased. Treatment of SHRs with sesamin dose-dependently reversed these biochemical and molecular abnormalities in aortas. CONCLUSION: Long-term treatment with sesamin improves arterial function in SHR through the upregulation of eNOS expression and downregulation of p22(phox) and p47(phox) expression.
Subject(s)
Aorta, Thoracic/physiopathology , Dioxoles/pharmacology , Down-Regulation/genetics , Lignans/pharmacology , NADPH Oxidases/antagonists & inhibitors , Nitric Oxide Synthase Type III/biosynthesis , Up-Regulation/genetics , Animals , Aorta, Thoracic/drug effects , Aorta, Thoracic/enzymology , Dioxoles/therapeutic use , Down-Regulation/drug effects , Hypertension/drug therapy , Hypertension/enzymology , Hypertension/physiopathology , Lignans/therapeutic use , Male , NADPH Oxidases/biosynthesis , Nitric Oxide Synthase Type III/genetics , Nitric Oxide Synthase Type III/physiology , Organ Culture Techniques , Protein Subunits/antagonists & inhibitors , Protein Subunits/biosynthesis , Random Allocation , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Up-Regulation/drug effectsABSTRACT
This study was designed to evaluate the in vivo cardioprotective effects of food-derived sesamin in spontaneously hypertensive rats (SHR). The study was performed with 17-week-old male normotensive Wistar-Kyoto rats (WKY) and SHR which are untreated or treated with orally administered sesamin for 16 weeks before they were sacrificed. Long-term treatment with sesamin obviously improved left ventricular (LV) hypertrophy and fibrosis in SHR, as indicated by the decrease of LV weight/body weight, myocardial cell size, cardiac fibrosis and collagen type I expression as well as the amelioration of the LV ultrastructure. These effects were associated with reduced systolic blood pressure, enhanced cardiac total antioxidant capability and decreased malondialdehyde content, nitrotyrosine level and transforming growth factor ß1 (TGF-ß1) expression. All these results suggest that chronic treatment with sesamin improves LV remodeling in SHR through alleviation of oxidative and nitrative stress, reduction of blood pressure and downregulation of TGF-ß1 expression.
Subject(s)
Antihypertensive Agents/pharmacology , Dioxoles/pharmacology , Lignans/pharmacology , Ventricular Remodeling/drug effects , Animals , Blood Pressure/drug effects , Collagen Type I/genetics , Collagen Type I/metabolism , Disease Models, Animal , Down-Regulation , Fibrosis/drug therapy , Fibrosis/pathology , Heart/drug effects , Heart/physiopathology , Hypertension/drug therapy , Hypertension/pathology , Hypertrophy, Left Ventricular/drug therapy , Hypertrophy, Left Ventricular/pathology , Male , Malondialdehyde/blood , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/pathology , Myocytes, Cardiac/ultrastructure , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Inbred SHR , Rats, Inbred WKY , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolism , Tyrosine/analogs & derivatives , Tyrosine/bloodABSTRACT
OBJECTIVE: To study the influence of Secoisolariciresinol Diglucoside (SDG) combined with Bortezomib on induction of apoptosis in lung cancer cell line A549 and its relative mechanisms. METHODS: The effect on proliferation was evaluated by MTT assay. The cell apoptosis was studied by flow cytometry and Hoechst 33342 staining. Colorimetric method was used to detect the activity of Caspase-3. Real time PCR was used to detect the expression of Caspase-3, BCL-2 and BAX mRNA. Western blot was used to determine the change of p-JNK, BCL-2 and BAX protein expression in A549 cells. RESULTS: The cell growth was significantly slowed down and the cell apoptosis was induced after the combined treatment. Meanwhile the Caspase-3 activity and the expression of Caspase-3 mRNA were obviously increased, the expression of BCL-2 mRNA and protein were significantly down regulated and the expression of BAX. p-JNK mRNA and protein were significantly up regulated after the combined treatment. CONCLUSION: The results demonstrate that SDG combined with Bortezomib can significantly induce apoptosis of A549 cells, its mechanisms may be involved in activation of the JNK pathway.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Boronic Acids/pharmacology , Butylene Glycols/pharmacology , Glucosides/pharmacology , Lung Neoplasms/pathology , Protease Inhibitors/pharmacology , Pyrazines/pharmacology , Antineoplastic Agents/administration & dosage , Boronic Acids/administration & dosage , Bortezomib , Butylene Glycols/administration & dosage , Caspase 3/genetics , Caspase 3/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Drug Therapy, Combination , Flow Cytometry , Glucosides/administration & dosage , Humans , Protease Inhibitors/administration & dosage , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Pyrazines/administration & dosage , RNA, Messenger/genetics , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , bcl-2-Associated X Protein/genetics , bcl-2-Associated X Protein/metabolismABSTRACT
This study is to observe anti-lipotoxic effect of sesamin on renovascular hypertensive rats fed with a high-fat, high-sucrose diet. Thirty-four complex model rats were induced by two-kidney, one-clip method and on high-fat and refined-carbohydrate diet for thirteen weeks. From the fifth week, intragastric administration of sesamin (120, 60 and 30 mg x kg(-1) x d(-1)) lasted for eight weeks. Blood pressure (BP), blood fat (BF), blood glucose (BG), free fatty acids (FFA), insulin (Ins), tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 were determined. Pathological changes of pancreas, perirenal fat and liver were semiquantitatively analyzed. In sesamin (120 and 60 mg x kg(-1) x d(-1)) group, it was found that there were decrease of levels of BP, BF, BG, TNF-alpha, IL-6 and FFA, improvement of insulin resistance and glucose tolerance, alleviation of body weight, humid weight of fat, liver and pancreas and their organ index, and reduction of islet cell hyperplasia and amount of lipid droplet vacuoles in lipocyte and hepatocyte. It is implied that sesamin had anti-lipotoxic effect and its mechanism may be closely associated with the amelioration of insulin resistance via reducing lipidoses in hepatocyte and inflammatory adipokines such as TNF-alpha and IL-6.
Subject(s)
Anticholesteremic Agents/pharmacology , Antihypertensive Agents/pharmacology , Diet, High-Fat , Dioxoles/pharmacology , Hypertension, Renovascular/blood , Insulin Resistance , Lignans/pharmacology , Adipocytes/drug effects , Animals , Anticholesteremic Agents/administration & dosage , Antihypertensive Agents/administration & dosage , Blood Glucose/metabolism , Blood Pressure/drug effects , Body Weight/drug effects , Cholesterol/blood , Dioxoles/administration & dosage , Dose-Response Relationship, Drug , Fatty Acids, Nonesterified/blood , Glucose Tolerance Test , Hypertension, Renovascular/pathology , Insulin/blood , Interleukin-6/blood , Islets of Langerhans/pathology , Lignans/administration & dosage , Liver/pathology , Male , Pancreas/pathology , Rats , Rats, Sprague-Dawley , Sucrose , Triglycerides/blood , Tumor Necrosis Factor-alpha/bloodABSTRACT
In the present study, we aimed to evaluate the protective effect of sesamin on kidney damage and renal endothelial dysfunction in two-kidney, one-clip renovascular hypertensive rats fed with a high-fat-sucrose diet (2K1C rats on HFS diet). Sesamin was intragastrically administered to 2K1C rats on HFS diet for eight weeks. Then, we measured the levels of serum hydrogen peroxide (H2O2), total antioxidant capability (T-AOC), renal malonaldehyde (MDA), total-erythrocuprein (T-SOD) and glutathione peroxidase (GSH-P(X)). The expressions of endothelial nitric oxide synthase (eNOS), nitrotyrosine and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit p47(phox) in the left and right renal cortexes were detected by Western blotting. Pathological changes in the left and right renal cortexes were observed by periodic acid-schiff staining (PAS) and Masson's staining. Treatment with sesamin (120 and 60mg/kg⻹·d⻹) in 2K1C rats on HFS diet improved renal function, corrected structural abnormalities, and attenuated renal oxidative stress. Furthermore, sesamin increased eNOS protein expression and reduced nitrotyrosine and p47phox protein expression. These results demonstrated that long-term treatment with sesamin had renoprotective effect and improved renal endothelial dysfunction via upregulation of eNOS expression and reduction of NO oxidative inactivation in both clipped and contralateral kidneys of 2K1C rats on HFS diet, and sesamin may have a favorably therapeutic value in treating chronic kidney disease in patients with hypertension and hyperlipemia.