Effect of N-acetyl-L-cystein on blood pressure and endothelial function in aorta of rats exposed to chronic intermittent hypoxia

AIM: To investigate the effect of N-acetyl-L-cystein (NAC) on blood pressure and endothelial function in the aorta of the rats exposed to chronic intermittent hypoxia (CIH). METHODS: Thirty healthy male SD rats were randomly divided into 3 groups: control group, CIH group and CIH+NAC group. The systolic blood pressure (SBP) was measured with tail-cuff me-thod. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression of endothelial nitric oxide synthase (eNOS） and endothelin-1 (ET-1) in the thoracic aorta. The protein expression of eNOS in the thoracic aorta was examined by Western blotting. The levels of ET-1 in the thoracic aorta and serum were detected by radioimmunoassay. The serum nitric oxide was determined by nitric acid reduction method.The superoxide dismutase (SOD) activity in peripheral blood plasma was detected by xanthine oxidase method.The serum malondialdehyde content was detected by thiobarbituric acid method, and superoxide anion (O-·2) in thoracic aorta was determined by chemical colorimetric method. RESULTS: Compared with the control animals, CIH exposure was associated with decreased SOD level, and NAC-treated CIH animals showed recovery in SOD level. NAC treatment prevented CIH-induced hypertension as well as CIH-induced increase in MDA. The aorta eNOS mRNA and protein, and serum NO levels in CIH group were lower than those in control group, and those in NAC treatment group were higher than those in CIH group. The increases in ET-1 mRNA,ET-1 protein and O-·2 levels in the aorta, and the elevated circulating ET-1 level were also observed in CIH-exposed animals. Treatment with NAC significantly decreased the mRNA and protein levels of ET-1, the O-·2 content, and the circulating ET-1 level in CIH-exposed animals. CONCLUSION: NAC protects endothelial function and alleviates hypertension by suppressing the oxidant stress in the aorta tissues, indicating that oxidant stress may be involved in the mechanism of endothelial disorder of CIH-induced hypertension.     

Effects of LPS on expressions of ET-1, eNOS and iNOS mRNA in human umbilical vein endothelial cells

AIM: To observe the direct effect of LPS on expressions of ET-1, eNOS, and iNOS mRNA in human umbilical vein endothelial cells, and further research the molecular mechanism of effect of LPS on production of ET-1 and NO. METHODS:The third passage of cultured human umbilical vein endothelial cells was incubated with low concentration (100 μg/L) of LPS for 6 h. Total RNA was extracted. The expressions of ET-1, eNOS, and iNOS mRNA were analyzed by semi-quantitative RT-PCR method. RESULTS: ET-1 mRNA experession increased significantly, while expression of eNOS mRNA decreased significantly, and there was no significant change in expression of iNOS mRNA. CONCLUSION: In human umbilical vein endothelial cells, low concentration of LPS enhanced the expression of ET-1 mRNA, inhibited the expression of eNOS mRNA, and had no significant effect on the expression of iNOS mRNA.     

Impact of hydrogen sulfide donor on endothelin-1 and connective tissue growth factor expression in rats with pulmonary hypertension induced by high pulmonary blood flow

AIM: To explore the possible impact of hydrogen sulfide (H2S) donor-sodium hydrosulfide (NaHS) on endothelin-1 (ET-1) and connective tissue growth factor (CTGF) expressions in rats with pulmonary hypertension induced by high pulmonary blood flow. METHODS: Thirty-two male SD rats were randomly divided into 4 groups: shunt group, shunt+NaHS group, sham group and sham+NaHS group. Rats in shunt group and shunt+NaHS group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. After 11 weeks of experiment, rat systolic pulmonary artery pressure (SPAP), lung tissue H2S, plasma ET-1 concentration and lung tissue ET-1mRNA expression, as well as pulmonary artery CTGF protein expression were detected.RESULTS: After 11 weeks of experiment, SPAP, lung tissue ET-1mRNA, plasma ET-1 as well as pulmonary artery CTGF expressions were increased markedly, respectively, whereas H2S in lung tissue decreased significantly in rats of shunt group as compared with that in sham group (all P<0.05). After administration of NaHS for 11 weeks, H2S in lung tissue increased significantly, whereas SPAP, plasma ET-1 and lung tissue ET-1 mRNA expression as well as pulmonary artery CTGF protein expression decreased significantly, respectively, in rats of shunt+NaHS group as compared with that in shunt group (all P<0.05).CONCLUSION: NaHS might be involved in the development of pulmonary hypertension induced by high pulmonary blood flow by down-regulating vasoactive peptides ET-1 and CTGF expressions in lung tissues of rats.     

The dynamic changes of plasma nitric oxide and endothelin-1 in prehepatic portal hypertension rats

AIM:To observe the dynamic changes of plasma levels of nitric oxide(NO) and endothelin (ET-1) in portal veins of the rats during prehepatic portal hypertension, and investigate the role of them in hyperdynamic circulation.METHODS:The models of prehepatic portal hypertension were established in Sprague-Dawley rats by means of partial portal vein ligation (PVL). The plasma levels of nitrite/nitrate (NO₂^-/NO₃^-) and ET-1 in the portal veins were detected by the method of nitric reductase and radioimmunoassay, respectively. In this study, rats were divided into normal, sham operation (SO) and PVL group. SO and PVL rats were divided into several subgroups according to different time after operations. Meanwhile, the changes of several hemodynamic indexes in these rats were also measured.RESULTS:The levels of NO₂^-/NO₃^- were significantly increased and ET-1 were significantly decreased in rats at different time after PVL compared with normal control, whereas the hemodynamic indexes changed accordingly.CONCLUSION:The portal hypertensive rats are in hyperdynamic circulatory state (HCS). NO and ET-1 may play an important role in the induction and maintenance of HCS.     

Protection of the limb ischemic preconditioning on the injury of gut is concerned with NO/ET-1 system in rats

AIM: To study the relationship between disturbance of nitric oxide/endothelin-1 (NO/ET-1) and the injury of gut following limb ischemia-reperfusion (I/R) in rats as well as the regulation of NO/ET-1 system by limb I/R preconditioning (IPC). METHODS: A limb ischemia-reperfusion injury model in rats was established. The animals were randomly divided into three groups: control group, IR group and IPC group. The contents of diamide oxidase(DAO), nitric oxide (NO), endothelin-1 (ET-1) and ratio of nitric oxide/endothelin-1 (NO/ET-1) in the plasma and the gut were measured. The leavels of myeloperoxidase, ratio of DNA chain (%), total nitric oxide synthase (tNOS), inducible nitric oxide synthase (iNOS) and constitutive nitric oxide synthase (cNOS) in the gut were determined. The expression of iNOS and endothelial NOS (eNOS) were detected by the immunohistochemical method. RESULTS: It was found that the levels of NO, ET-1 in the plasma and the gut tissue all increased after reperfusion, while the values of NO/ ET-1 decreased. The values of DAO in the plasma and MPO in the gut increased, while the contents of DAO and the ratio of DNA chain (%) in the gut decreased. The expression of iNOS elevated, cNOS (mainly eNOS) reduced and total NOS increased. The protection of the limb IPC attenuated the disturbance of NO/ET-1. CONCLUSION: The intestinal injury following limb I/R is related to the disturbance of NO/ET-1. The protection of the limb IPC might be conducted by its regulating NO/ET-1 system. The endothelial NOS increases and non-endothelial NOS decreases in this situation.     

Effect of caveolin-1 expression on high-salt diet-induced endothelial dysfunction in type 1 diabetic rats

AIM: To investigate the underlying mechanisms responsible for endothelial dysfunction of type 1 diabetes mellitus (DM) rats fed with high-salt diet. METHODS: Type 1 DM was induced by intraperitoneal injection of streptozotocin (70 mg/kg). Normal and diabetic rats were fed high-salt food (HS, 8% NaCl) and standard food for 6 weeks, respectively. Isometric tension of the mesenteric arteries were measured. The expression of Akt, endothelial nitric oxide synthase (eNOS) and caveolin-1 (Cav-1) was examined by Western blot. RESULTS: The rats in DM+HS group exhibited more pronounced impairment of vasorelaxation to acetylcholine and insulin compared with either DM group or HS group (P<0.01). Akt and eNOS phosphorylation levels, and nitric oxide (NO) concentration in DM+HS group were significantly lower than those in DM group (P<0.01). The level of Cav-1 in DM+HS group was significantly higher than that in DM group and HS group. CONCLUSION: Impaired endothelial Akt activation, increased Cav-1 expression and resultant decreased eNOS activation contribute to aggravate high-salt diet-induced endothelial dysfunction and hypertension in DM rats.     

Effect of shenmai injection on myocardial eNOS mRAN expression in myocardial ischemic rats

AIM:To study the effect of shenmai injecti on (SMI),a Chinese medicine,on nitric oxide and the expression of endothelial nitric oxide synthase (eNOS) mRNA in myocardium of rats with experimental myocar dial ischemia.METHODS:Rats were randomly divided into four groups:control,m odel (ischemia),SMI 1 and SMI 2 group.A rat model of acute myocardial ischemia was established by isoprenaline treatment and the lift of ST segment in ECG was used as the index of myocardial ischemia.The nitric oxide (NO) contents in ser um and myocardium and the expression of eNOS mRNA in myocardium were measured.RESULTS:Compared with control group,ST segment of ECG was sign ificantly elevated 20,30,40 min after myocardial ischemia in model group,the lift peak of ST segment occurred 20 min after myocardial ischemia,the concentra tion of NO in the serum and myocardium and the expression of eNOS mRNA in myocar dium were significantly lowered in model group.Compared with the model group,i n SMI 1 group and SMI 2 group,the concentration of NO in the serum and myocard ium and the expression of eNOS mRNA in myocardium were significantly increased,the lift of ST segment were significantly reduced 20,30,40 min after myocardi al ischemia.Compared with SMI 1 group,the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium and the lift of ST seg ment were not statistically different in SMI 2 group.CONCLUSION:Shenmai injection can increase the expression of eNO S mRNA in myocardium and the content of NO,and protect against myocardial ische mia in rats.     

Effect of diltiazem on heme oxygenase-1 and nitric oxide synthase in rats with pulmonary hypertension

AIM: To investigate the action of diltiazem (a calcium antagonist) on the expression of heme oxygenase (HO) -1 and nitric oxide synthase (NOS) in the small pulmonary arteries (SPA) of rat in chronic hypoxia. METHODS: Chronic pulmonary arterial hypertension models were established by treating the rats in hypoxic environment for 6 weeks. After 2 weeks of hypoxia, rats were treated with diltiazem (15 mg/kg/day). Right ventricular systolic pressure (RVSP) and right ventricular hypertrophy index (RVHI) were measured. Pathological changes in the lungs were observed under the light microscope and transmission electron microscope. The expression and distribution of heme oxygenase (HO) -1, endothelial NOS (eNOS) and inducible NOS (iNOS) were tested by immunohistochemistry and Western blot. Guanosine-3', 5'-cyclic monophosphate (cGMP) of lung tissues were detected with radioimmunoassay. RESULTS: Diltiazem significantly decreased abnormal RVSP, and RVHI in model rats, attenuated the SPA media thickeness, and recovered abnormal eNOS and iNOS expression in SPA. Whereas diltiazem had little effect on the increased HO-1 expression in SPA caused by hypoxia and ultrastructure injury in endothelium. cGMP levels were corresponded with HO-1. CONCLUSION: Diltiazem has a significant effect on inhibiting hypoxic pulmonary hypertension structural remodeling. These effects might be partly attributed to the suppression of iNOS, promotion of eNOS, and not attenuation HO-1 expression in the lung of hypoxic rats.     

Protective effect of famotidine against gastric mucosa damage resulting from interdicted hepatic portal in the liver cirrhosis rat

AIM: To investigate the influence of hepatic portal interdicting on gastric mucosa of the liver cirrhosis rats and the protective effect of famotidine on gastric mucosa. METHODS: Fifty Wistar rats were randomly divided into 5 groups; group A (healthy rats), group B (healthy rats with interdicted hepatic portal), group C (liver cirrhosis rats without interdicted hepatic portal), group D (liver cirrhosis rats with interdicted hepatic portal) and group E (liver cirrhosis rats without interdicted hepatic portal and use of famotidine); The gastricfluid pH, gastric mucous content (GMC), gastric mucosa blood flow (GMBF) and the damage index of gastric mucosa (DIGM) of the every group were observed. RESULTS: There is no significant difference in the above parameters between group A and group B (P>0.05). Compared with group A, GMBF of group C, D, and E were reduced (P<0.01), pH, GMC of group C, D and E were also reduced (P<0.05); DIGM of group C, D and E were increased (P<0.01). All the changes of group D were more obvious than those of group C (P<0.01); While there was no significant difference in all the parameters between group E and group C (P>0.05). CONCLUSION: Hepatic portal interdicting can aggravate the gastric mucosa damage of the liver cirrhosis rat, and famotidine can protect against such gastric mucosa injury through improving the microcirculation of gastric mucosa.     

Effect of cilazapril on pulmonary vascular endothelial dysfunction in hypoxic rats

AIM:To study the effect of cilazapril on pulmonary vascular endothelial dysfunction in hypoxic rats. METHODS:The structure and function of endothelium in hypoxic rats were studied by biochemical analysis, radioimmunoassay, transmission electron microscope and correlated with hemodynamic. RESULTS:1) The change and damage of ultrastructure in endothelial cell (EC) were obsevered in hypoxic rats. 2) The contents of plasma nitric oxide (NO) and superoxide dismutase (SOD) activity in blood as well as endothelial nitric oxide synthase (eNOS) activity in the lung tissue were significantly lower in the hypoxic rat than those in contral animals. The concentrations of plasma endothelin-1(ET-1) and angiotensin converting enzyme(ACE) as well as malondialdehyde(MDA) were significantly higher in the hypoxic rat than these in contral animals. The relaxing and contracting factors had a significant positive/negative correlation with mean pulmonary artery pressure (mPAP). 3) Cilazapril significantly decreased the level of ET-1 and ACE and significantly increased the level of NO and activity of eNOS and SOD. At the same time, cilazapril extenuated hypoxia-induced injuries of EC. CONCLUSION:The results indicate that damaging structure and dysfunction of EC existes in hypoxic rats. The cilazapril effectively preventes and treates the chronic hypoxic PH by relieving the injury and improving secretion in EC.     

Role of nitric oxide in proliferation and secretion effect of vascular endothelial cells induced by vascular endothelial growth factor

AIM: To investigate the role of nitric oxide in proliferation and secretion of vascular endothelial cells induced by vascular endothelial growth factorr (VEGF). METHODS: The in vitro cultured vascular endothelial cells of rabbit aorta were divided into control group, VEGF-treated group and VEGF+L-NAME treated group, the absorbance (A) value of vascular endothelial cells, endothelin-1(ET-1) and von Willebrand factor (vWF) in the supernatant were examined by WST-1 assay, radioimmunoassay and ELISA. RESULTS: The A value in VEGF and VEGF+L-NAME treated group were higher than that in control group (P＜0.01). A value in VEGF group was higher than that in VEGF+L-NAME group the ET-1 and vWF were markedly decreased in VEGF group compared with the control and VEGF+L-NAME treated group (P＜0.05, P＜0.01). These results indicated that VEGF promoted the proliferation and inhibited the secretion of ET-1 and vWF in vascular endothelial cells, and L-NAME inhibited the effect of VEGF. CONCLUSION:Nitric oxide is an important mediator in the process of stimulating proliferation and regulating secretion of vascular endothelial cells by VEGF.     

Hyperhomocysteinemia induces endothelial dysfunction and aggravates microcirculation dysfunction and microthrombosis

AIM: To establish a microthrombus model by carrageenan (Ca)/ lipopolysaccharides (LPS) intraperitoneal injection in rats with hyperhomocysteinemia (HHcy) and endothelial dysfunction induced by L-methionine intake.METHODS: ① Male Sprague Dawley rats were randomly divided into 2 groups: control and endothelial dysfunction (HHcy) groups. L-methionine was administered by gavage in HHcy group for total 4 weeks.Purified water was administered by gavage in control rats.Plasma Hcy,NO and vWF were examined and the thoracic aorta were excised after 4 weeks of L-methionine treatment to evaluate endothelial function.② Male Sprague Dawley rats were randomly divided into 3 groups to establish a microthrombus formation model with Ca/ LPS: control,microthrombus formation (Ca/LPS) and endothelial dysfunction plus mitoarothrombus formation (HHcy＋Ca/LPS) groups.Control rats were injected with normal saline (NS).Ca/LPS rats were intraperitoneally injected with carrageenan (Ca) and followed by lipopolysaccharides (LPS) 16 h later.HHcy＋Ca/LPS rats were intragastric gavaged by L-methionine for total 4 weeks,and then were injected with Ca/LPS in the same way as Ca/LPS group.Cruor parameters and platelet count were detected at 20 h after LPS or NS injection and the mesentery microcirculation was monitored.Plasma NO and vWF were also detected at 24 h after LPS or NS injection.RESULTS: ① Plasma Hcy concentrations and vWF level were significantly increased in HHcy group,while plasma NO content was significantly decreased compared with that in control group.Endothelial dependent relaxation (EDR) of aortic rings was significantly decreased in HHcy group,suggesting endothelial damage/dysfunction was induced by HHcy.② Mesentery capillary was obviously blocked by microthrombus in Ca/LPS rats and was blocked more seriously in HHcy＋Ca/LPS rats.Cruor parameter results suggested that Ca/LPS rats were in hypercoagulable phase and HHcy＋Ca/LPS rats were in hypocoagulable phase at 20 h after LPS injection.Platelet count and plasma NO content in HHcy＋Ca/LPS group were significantly decreased,while plasma vWF level was significantly increased compared with Ca/LPS group.CONCLUSION: L-methionine intake induces severe HHcy and causes endothelial dysfunction in rats.Microcirculation dysfunction and microthrombosis can be caused by Ca/LPS intraperitoneal injection and may be aggravated by endothelial dysfunction.     

Effect of tissue kallikrein gene treatment on blood pressure in type 2 diabetic rats and its mechanism

AIM:To study the effect of tissue kallikrein gene (HK) treatment on blood pressure in type 2 diabetic rats and its mechanism. METHODS:Male Wistar rats were injected with low dose streptozotocin and fed with diets enriched in fat and sugar to form type 2 diabetic model. Recombinant adeno-associated viral vectors (rAAV)-mediated HK gene (HK group) or LacZ gene (LacZ group) was introduced to the diabetic rats. The systolic blood pressure was measured every 2 weeks. The acetylcholine (Ach)-dependent vasodilation response, the synthesis of nitric oxide (NO), the expression of endothelin-1 (ET-1) and endothelin-A receptor (ETA-R) in the aorta were detected. RESULTS:(1) Systolic blood pressure was significantly higher in diabetic rats than that in normal control rats. In HK group, systolic blood pressure was significantly reduced within 2 weeks after injection with rAAV·HK, reached near normal levels at 4 weeks and kept until the experiments ended (16 weeks). (2) In LacZ group, Ach-dependent vasodilation response of isolated aorta was markedly decreased than that in HK group (P<0.01). (3) The concentration of NO in the aorta of HK group were significantly higher than those in LacZ group. The expression of ET-1 and ETA-R mRNA were significantly decreased in HK group compared with those in LacZ group (P<0.01). CONCLUSION:rAAV-mediated HK gene delivery efficiently lowed blood pressure and attenuated the endothelial function partly through increasing the concentration of NO and inhibiting the expression of ET-1 and ETA-R of aorta in type 2 diabetic rats.     

Effects of macrophage polarization during development of liver cirrhosis in rats

AIM: To explore the state of macrophage polarization and its relation with intestinal endotoxemia-endoplasmic reticulum stress in the development of liver cirrhosis induced by multiple pathogenic factors in rats. METHODS: The male SD rats (n=36) were randomly divided into normal control group and liver cirrhosis model group, and sacrificed at the end of the 4th, 6th and 8th weeks. The rat model of liver cirrhosis was induced by multiple pathogenic factors. The levels of alanine aminotransferase (ALT), endotoxin, homocysteine (Hcy) in the plasma, and inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), arginase-1 (Arg-1) and interleukin-10 (IL-10) in the liver tissues were detected by ELISA. Histopathological change of the liver was observed under microscope with the staining of hematoxylin and eosin (HE) and van Gieson (VG). The expression of glucose-regulated protein 78 (Grp78), nuclear factor-kappa B (NF-κB), interferon-regulatory factor 5 (IRF5), CD86, CD206 and transforming growth factor-β1 (TGF-β1) at mRNA levels in the liver tissues were detected by the method of real-time fluorescence quantitative PCR.RESULTS: Compared with the corresponding normal control group, the levels of ALT, endotoxin, Hcy in the plasma and Grp78 mRNA in the liver tissues in liver cirrhosis model group were significantly and gradually increased (P<0.05). The mRNA expression of NF-κB, IRF5 and CD86, and the protein levels of iNOS, TNF-α and IL-6 in the liver tissues were significantly increased (P<0.05), and they successively increased from the 4th week to the 6th week and decreased reversely at the 8th week. The mRNA expression of CD206, TGF-β1, Arg-1 and IL-10 in the liver tissues were significantly increased from the 6th week to the 8th week (P<0.05), and no significant difference at the 4th week was observed. The level of endotoxin in the plasma was correlated with the mRNA expression of Grp78 in the liver tissues (P<0.01). Both endotoxin in the plasma and Grp78 mRNA in the liver tissues were correlated with the mRNA expression of CD86 and CD206 in the liver tissues (P<0.01).CONCLUSION: The pathway of liver damage-intestinal endotoxemia-endoplasmic reticulum stress-macrophage polarization may be critical in the pathogenesis of liver cirrhosis induced by multiple pathogenic factors.     

Expression and significance of fgl2 in a model of rat coronary microthrombosis in situ

AIM: To explore the expression and significance of fibrinogen-like protein 2 prothrombinase/fibroleukin (fgl2) in a model of rat coronary microthrombosis. METHODS: Seventy-eight Sprague-Dawley male rats were randomly assigned to model group (n=36), sham operation group (n=36) for different time (1 h, 1 d, 1week, 2 weeks, 3 weeks, 4 weeks)(n=6, respectively), and normal control group (n=6). After closing aorta, sodium laurate was injected into aortic root of the animals in model group. The animals in sham operation group were injected with normal saline for control. Microthromb were detected with HE staining and modified martius-sarlet-blue stains (MMSB). von Willebrand factor (vWF) was measured with ELISA. fgl2 mRNA and protein expressions were detected by RT-PCR and immunohistochemistry staining, respectively. RESULTS: Microthromb were observed in rat coronary microvasculature. vWF was increased in plasma. fgl2 was induced in model group compared to sham operation group (P<0.05), no significant changes were observed in sham operation group and normal control group (P>0.05). CONCLUSION: The functional disorder of endothelial cells induces the expression of fgl2 in the coronary microvasculature. fgl2 prompts the thrombosis with platelet and fibrin.     

Changes of plasma endothelin-1 and von Willebrand factor in patients undergoing cerebral digital substration angiography

AIM: To investigate the effects of cerebral digital substration angiography (DSA) on the expression of endothelin-1(ET-1) and von Willebrand factor(vWF) in vascular endothelial cells from the patients with ischemic cerebrovascular disease. METHODS: The plasma levels of ET-1 and vWF were measured by enzyme-linked immunosorbent assay in 108 patients before, immediately and 24 h after cerebral DSA. RESULTS: The plasma level of endothelin-1 significantly increased immediately and 24 h after cerebral DSA as compared with that before the procedure (P<0.05). The level of vWF was not changed before and after operation. CONCLUSION: The marked and sustained increase in endothelin-1 plasma level after cerebral DSA is probably related to mechanical endothelial injury by the procedure of cerebral DSA.     

Role of SIRT1/eNOS/NO signaling pathway in protective effect of ginsenoside Rb1 against replicative senescence of endothelial cells

AIM: To explore the effect of ginsenoside Rb1 on replicative senescence of endothelial cells and the role of SIRT1/eNOS/NO signaling pathway in this process. METHODS: The replicative senescence model of primary human umbilical vein endothelial cells (HUVECs) was established. The morphological change of the cells, the proportion of senescence-associated β-galactosidase (SA-β-Gal) positive cells and the plasminogen activator inhibitor 1 (PAI-1) expression were detected to assess the senescence model. The expression of eNOS and PAI-1 at mRNA and protein levels in the aging cells was determined by real-time PCR and Western blot before and after silencing of SIRT1 was performed. The NO concentration in the cell culture supernatant was measured by nitrate reductase assay. RESULTS: HUVECs with cumulative population-doubling level (CPDL) at 16 were chosen as the replicative senescence model in this research. Ginsenoside Rb1 at 80 μmol/L significantly reduced the expression of PAI-1 at mRNA and protein levels. Furthermore, ginsenoside Rb1 increased the expression of SIRT1 and eNOS at mRNA and protein levels, and increased the NO content. SIRT1 silencing inhibited the expression of eNOS at mRNA and protein levels and reduced NO generation, leading to an increase in the expression of PAI-1 at mRNA and protein levels. Upon intervention of ginsenoside Rb1, the eNOS and PAI-1 expression and the level of NO were not reversed. CONCLUSION: Ginsenoside Rb1 modulates SIRT1/eNOS/NO signaling pathway to prevent the replicative senescence of HUVECs.     

Effect of angiontensin-(1-7) on development of monocrotaline induced pulmonary arterial hypertension and vascular remodeling

AIM: To investigate the effect of angiotensin-(1-7) ［Ang-(1-7)］ on the development of monocrotaline (MCT) induced pulmonary arterial hypertension and vascular remodeling.METHODS: 60 Sprgue-Dawely rats were randomly assigned into three groups: control group, PAH group and PAH+Ang-(1-7) group. Rats in PAH group and PAH +Ang-(1-7) group received 60 mg/kg MCT injection subcutaneously and after 24 h received either saline or 24 μg·kg-1·h-1 of Ang-(1-7) injection via osmotic minipumps for 4 weeks. These rats in control group were firstly injected saline subcutaneously and then received saline injection via osmotic minipumps.RESULTS: After 4 weeks, in PAH group, right ventricular systolic pressure (RVSP), right ventricular hypertrophy index (RVHI), percentage of wall thickness (WT%) and percentage of wall area (WA%) of pulmonary artery were significantly increased and NO concentration, the level of endothelial nitric oxide synthase (eNOS), eNOS Ser1177-phosphorylation were significantly decreased compared with control group. However, RVSP, RVHI, WT %, WA % were dramatically decreased in PAH +Ang-(1-7) group and NO concentration, the level of eNOS protein, eNOS Ser1177-phosphorylation were significantly increased compared with PAH group.CONCLUSION: These results suggest that Ang-(1-7) could prevent the development of monocrotaline induced pulmonary arterial hypertension and vascular remodeling, which appears to be associated with up-regulation of NO concentration and the level of eNOS protein, eNOS Ser1177-phosphorylation.     

Changes of portal hemodynamics in the development of experimental liver cirrhosis

AIM: To understand the formation of portal hypertension through the change of portal hemodynamics on experiment cirrhosis. METHODS: Carbon tetrachloride was subcutaneously injected in the rat. The changes of the portal hemodynamics in the pathological process of liver tissue were observed. RESULTS: The liver underwent degeneration, necrosis of hepatocytes, and the normal architecture of the liver lobules was replaced by pseudolobule, which consist of regenerative hepatocytes and fibrous septa. The diameter, the blood flow velocity and the blood flow quantity of portal were significantly higher than that in former group (P＜0.05 or P＜0.01) two weeks after the injection of carbon tetrachloride. In the fifteenth week, these parameters were lower than that before owing to the forming of portacaval collateral circulation (P＜0.01). The congest index of the portal in second week, fifth week and fifteenth week were statistically higher than its predecessor (P＜0.05 or P＜0.01), except that in tenth week, which had no statistical significance (P>0.05). CONCLUSION: The changes in hemodynamics of the portal are in accordance with the changes in pathology of liver in the formation of liver cirrhosis.     

Effects of eplerenone on expression and activity of aortic endothelial nitric oxide synthase in hypertensive rats induced by high-salt intake

AIM: To explore the effects of eplerenone on the expression and activity of aortic endothelial nitric oxide synthase(eNOS) in high salt-induced hypertensive rats.METHODS: Male Wistar rats(4 week old, weighting 50～60 g) were randomly divided into control group, high-salt diet group and eplerenone group. The rats in control group were fed with ordinary rodent animal diet, the rats in high-salt group and eplerenone group were exposed to 5% salt diet for 16 weeks and administrated with the same dosage of saline or eplerenone(40 mg·kg^-1·d^-1) by gavage for 4 weeks, respectively. Systolic blood pressure(SBP) was measured by tail-cuff every 2 weeks. The rats were sacrificed after 16 weeks and the thoracic aorta was collected. The aldosterone content in the aorta was measured by ELISA. The protein levels of mineralocorticoid receptor(MR) and eNOS were determined by Western blot. The activitie of constitutive NOS(cNOS) was measured by chemocolorimetry. The protein localization of eNOS, neuronal nitric oxide synthase(nNOS) and MR was observed by immunohistochemistry.RESULTS: A process of 8-week high-salt diet increased SBP gradually. SBP in the rats exposure to high salt for 16 weeks was significantly higher than that in control group(P<0.05). After 4 weeks of eplerenone treatment, SBP in the rats was significantly lower than that before treatment(P<0.05). Compared with control group, the aldosterone content in the aorta were significantly increased in high-salt diet group and eplerenone group(P<0.05), the expression level of MR also increased significantly(P<0.05). Compared with control group, both eNOS protein expression(P<0.05) and cNOS activity in high-salt diet group were significantly decreased(P<0.05). The protein expression of eNOS as well as cNOS activity in aorta increased significantly in eplerenone group compared with high-salt diet group(P<0.05).CONCLUSION: Aldosterone content in aorta of high-salt-induced hypertensive rats increases significantly. Aldosterone attenuates the protein expression of eNOS and reduces the enzyme activity through the activation of mineralocorticoid receptor. The selective mineralocorticoid receptor antagonist eplerenone enhances the protein expression of eNOS and its activity, thereby improves eNOS function.