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<p><b>OBJECTIVE</b>To observe the inhibition of flavonoids extract (FE) from Inula britannica on oxidative stress in rat aorta after balloon injury.</p><p><b>METHOD</b>The model of vascular intimal hyperplasia was established by balloon injury. The rats were randomly divided into four groups: control, model, FE and captopril (CAP, positive control). The FE group was treated by intragastric administration with FE in dose of 12.5, 25, 50 mg x kg(-1) x d(-1). The level of malondialdehyde (MDA) and superoxide dismutase (SOD) in serum were detected by thiobarbituric acid (TAB) method and xanthine oxidase method respectively, and superoxide anion (O2-) in vessel was detected by dihydroethidium (DHE) staining. The histochemistry, immunohistochemistry and Western blot were used to observe the changes of appearance and SOD expression in vascular tissues after balloon injury.</p><p><b>RESULT</b>The best concentration of FE to rats was 50 mg x kg(-1) x d(-1). The neointima thickness in the model group was significantly higher than that in the FE group (50 mg x kg(-1) x d(-1)) and control group at 14 days after balloon injury (P < 0.01). The lever of MDA in serum of FE group was decreased (P < 0.01) and SOD was increased (P < 0.05) in both serum and vascular tissues. The level of O2*- in the drug group was lower than that in the model group.</p><p><b>CONCLUSION</b>FE can enhance the antioxidation capacities of vessel tissues by suppressing the formation of O2- induced by injury, by which FE inhibites neointima formation after balloon injury in rat.</p>
Subject(s)
Animals , Male , Rats , Antioxidants , Chemistry , Aorta , Metabolism , Blood Vessels , Metabolism , Catheterization , Down-Regulation , Ethidium , Blood , Metabolism , Flavonoids , Chemistry , Inula , Chemistry , Oxidative Stress , Random Allocation , Superoxide Dismutase , Blood , MetabolismABSTRACT
Objective To investigate the relationship between NF-?B activation and the expression of metastasis correlation factors.Methods The expression of NF-?B,COX-2,ICAM-1,VCAM-1 and MMP9 in colorectal cancer and normal tissues was detected using Western blotting.Results(1)The expression level of NF-?B was higher in cancer tissues than that in normal tissues(P
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Angiotensin Ⅱ(AngⅡ)can induce the expression of its precursor,angiotensinogen,in vascular smooth muscle cells(VSMC),which is related with increased activating protein-1(AP-1)binding to its cis-element located in the angiotensinogen gene promoter.In the present study,cycloheximide(CHX)was used as an inhibitor to interrupt c-Jun,the role of AP-1 in AngⅡ-induced its precursor gene activation was investigated by DNA-protein interaction and immunoblotting.The results showed that the level of c-Jun,the component of transcription factor AP-1,was significantly increased in the nucleus of VSMC after AngⅡ treatment.The majority of c-Jun was found in the nucleus but hardly detected in the cytoplasm by immunocytochemistry staining.Immunoprecipitation assays confirmed that AngⅡ could induce serine phosphorylation of c-Jun.EMSA results indicated that the level of phosphorylated of c-Jun had a positive correlation with AP-1 binding activity to cis-acting element of angiotensinogen gene and transcription activation of angiotensinogen.CHX inhibited AngⅡ-induced binding activity of AP-1 by reducing the phosphorylation of c-Jun,though it did not affect the expression of c-Jun.These findings suggest that the AP-1 phosphorylation induced by AngⅡ is one of the important mechanisms whereby AngⅡ regulates its precursor gene expression in feedback manner.It is found that CHX is an inhibitor to phosphorylation of c-Jun.
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The technique of genomics and proteomics is one of the fastest developments with the farest-reaching consequences in the high and new biotechnology in the world of today. It can be used to screen the target molecules of the action of traditional Chinese medicines, to identify the new effective components from traditional Chinese medicines, and to explore the mechanisms of the effects of traditional Chinese medicines. It meets the shortcomings of the conventional methodology being applied in the current studies of traditional Chinese medicine. Application of the theories and technique of genomics and proteomics in the study of traditional Chinese medicine would be of great significance for opening new research field of traditional Chinese medicine, for facilitating the integration of traditional Chinese medicine and modern biological science and technology, and for promoting the internationalization of traditional Chinese medicine.
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According to the theory of traditional Chinese medicine, blood stasis is a main pathological mechanism in the development of vascular diseases. Supplementing Qi and activating blood circulation, as the therapeutic principle for the blood stasis, has been adapted. Studies demonstrated that the supplementing Qi and activating blood circulation recipe could regulate the expression of vasoactive peptides in vascular smooth muscle cells (VSMCs). The recipe inhibited the formation of neointima following arterial intimal lesions through down-regulating expression of proliferation-related genes and phenotypic modulation of VSMCs. The recipe also markedly inhibited the adhesion and migration of VSMCs and matrix remodelling by means of a mechanism that balances extracellular matrix turnover rate. The purpose of this review is to summarize the recent advances made in our understanding of new functions for the recipe in regulating VSMCs behaviours and their microenvironment relevant to vascular diseases and maintaining proper homeostasis.
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AIM: To investigate whether the expression of integrin ?_3 and the activation of focal adhesion kinase(FAK) are involved in neointima formation after de-endothelium.METHODS: The model of intima hyperplasia was prepared by balloon injury.The levels of osteopontin(OPN),integrin ?_3 and FAK in vascular tissue were detected by Western blotting and immunohistochemistry.RESULTS: There were similar expression patterns in OPN,integrin ?_3 and FAK following balloon injury.The levels of three proteins were markedly increased 3 days after operation and reached the peak at 7th day.The increased FAK was mainly the phosphorylated form.The migration and proliferation of vascular smooth muscle cells was associated with the increase in the expression of integrin ?_3 and FAK,and was parallel with rapid turnover of extracellular matrix(ECM).CONCLUSION: The interaction of cells with ECM mediated by OPN and integrin ?_3 is essential for migration.The integrin ?_3-FAK pathway is involved in neointima formation.
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AIM: To determine the relationship between the changes of SM ?-actin and SM22? expression and vascular tone. METHODS: The expression of SM ?-actin and SM22? during restenosis after de-endothelialization were detected by Western blotting and immunohistochemistry. The structure of myofilaments was observed by transmission electron microscopy. The vascular contraction induced by phenylephrine was measured by a force transducer. RESULTS: The levels of SM ?-actin and SM22? expression in vascualar wall declined after de-endothelialization. The myofilaments in VSMC were modulated from well arranged and dense bundles to discrete network. However, the vascular tone and reactivity to agonist were much higher than that in control (P
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AIM: To study the effect of Salvia miltiorrhiza injection on rat atherosclerosis (AS), and elucidate the possible mechanism. METHODS: Wistar rats were fed with fat-rich diet and high dose of vitamin D_3 to induce AS, then treated with Salvia miltiorrhiza injection. Concentrations of triglyceride (TG) and total cholesterol (TC) in serum were measured by automatic serum biochemical assay. The level of ICAM-1 protein and mRNA were determined by Western blot and RT-PCR. RESULTS: Compared with the AS model group, the levels of TG and TC in serum were significantly lower in Salvia miltiorrhiza injection group (P
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Object To study the effective portion in Cornus officinalis Sieb. et Zucc. extract (COE) for the treatment of arrhythmia. Methods Effect of COE on chloroform induced ventricular fibrillation in mice and electrophysiology of isolated guinea pig papillary muscle were studied. Results Antiarrhythmic effect of COE may be related to its prolongation of action potential duration, increase of the absolute value of resting potential and a decrease of autonomy of sinus node. The effective portion in COE may be its total organic acid and a certain yet unknown trace substance, whereas its total glycosides were devoid of such activities. Conclusion Pharmacodynamic and myocardial electrophysiologic studies showed that the total organic acid and a certain unknown trace substance possessed the obvious antiarrhythmic activity.
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AIM: To study the relationship between expression of ? 3 integrin, focal adhesion kinase (FAK) and migration of vascular smooth muscle cells (VSMC) stimulated by osteopontin (OPN) and fibronectin (FN).METHODS: VSMC migration was examined using a model of wounding injury of confluent cultured cells. The expression of ? 3 integrin, FAK and Gax genes in VSMC were detected using Western blotting and RT-PCR. RESULTS: OPN and FN induced the migration activity of VSMC in a time-dependent manner ( P
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AIM: To investigate the relationship between NF-?B activation and intimal hyperplasia.METHODS: The protein extract from aorta was used to detect the expression of NF-?B p65,I?B?,ICAM-1 and COX-2 by Western blotting,and detection of threonine phosphorylation of p65 was performed by immunoprecipitation analysis.RESULTS: The level of p65 reached a maximum level at day 7,and then decreased significantly,which was higher than that in the control,in both cellular extract and nuclear extract at day 21 after balloon injury.The p65 level in cytoplasm had no obvious changes at different time points after balloon injury.The threonine phosphorylation of NF-?B p65 was negative correlated with the nuclear translocation of NF-?B.I?B? level showed a 15% decrease at day 1 after balloon injury,compared with control group,then returned at day 14,and reached to normal level at day 21.However,the expression of ICAM-1 and COX-2 was maximal at day 14,and then declined but higher than that in control group at day 21 after balloon injury.CONCLUSION: The activation of NF-?B p65 and the expression of ICAM-1 and COX-2 are parallel to the neointimal thickening after balloon injury.
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AIM: To investigate the mechanism of inhibiting the neointimal formation through blocking osteopontin (OPN) by using anti-OPN antibody. METHODS: The anti-OPN antibody was injected via tail vein after the left carotid artery of rats was de-endothelialized by balloon catheter. The expression of matrix metalloproteinase-2 (MMP-2), tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) and OPN were detected by gelatin zymogram, Western blotting, immunohistochemistry and flow cytometry. RESULTS: The neointima in rats treated with anti-OPN antibody was significantly thinner than that in untreated rats (P
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AIM: To investigate the effect of platelet-derived growth factor (PDGF) on ?_3 integrin gene expression and the role of ?_3 integrin on adhesion, migration and proliferation of vascular smooth muscle cells (VSMC) induced by PDGF. METHODS: ?_3 integrin gene expression was detected by RT-PCR. After ?_3 integrin extracellular domain was blocked, VSMC adhesion, migration and proliferation were measured by adhesion assay, a wound-culture model and [~3H]-TdR incorporation, respectively. RESULTS: After the interaction between ?_3 integrin and extracellular matrix was blocked, VSMC proliferation was inhibited in some degree and the rate of [~3H]-TdR incorporation into VSMC decreased 39%. The cell adhesion and migration were significantly inhibited when 10 mg/L anti-?_3 integrin antibody was added (P
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Objective To study the cardiomyocyte and renal cell apoptosis induced by overtraining, and the preventive effects of anisodamine against the adverse effects. Methods The rats were forced to swim till exhaustion to reproduce the animal model of overtraining. The animals were randomly divided into control group, exhausted group and anisodamine group. The exhausted group, depending on the recovery time after exhaustion, was divided again into exhaustion subgroup, 6h after exhaustion subgroup, and 24h after exhaustion subgroup. The animals in the anisodamine group received intraperitoneally 10mg/kg of anisodamine before the swimming overtraining, and divided again into 6h after anisodamine injection subgroup and 24h anisodamine injection subgroup. The cardiomyocyte and renal cell apoptosis was observed by the method of TUNEL, image analysis and flow cytometry. Results It was revealed by TUNEL that the number of apoptotic cardiomyocytes was increased after exhaustive swimming, especially in the 6h after exhaustion subgroup. The number of apoptotic cells was also increased in kidney of 0h, 6h and 24h after exhaustion subgroups, especially in 24h after exhaustion subgroup. The apoptosis ratio was also increased significantly in 0h, 6h and 24h after exhaustion subgroups as demonstrated by flow cytometry. Compared with the rats of exhausted group, the number of apoptotic cells in heart and renal tissue was decreased remarkably after anisodamine injection. Conclusion The apoptosis of cardiomyocyte and renal cell could be induced by exhaustive swimming. Compared with kidney, heart injury recovered more quickly. Anisodamine had the preventive effect on the injury to heart and kidney in exhausted rats.