Objective To observe the effect of ginsenoside Rb 1 and Re on cardiomyocyte apoptosis and expression of Bcl-2, Bax, Bad and Fas gene proteins after acute ischemia and reperfusion in rats and elucidate the possible mechanisms. Methods The ischemia-reperfusion heart model was made by ligating the left anterior descending branch of coronary artery in Wistar rats. The apoptotic cardiomyocytes were confirmed with transmission electron microscopy and counted with in situ nick labeling (TUNEL) method and light microscopy. The expression of Bcl-2?Bax?Bad and Fas gene proteins were studied by immunohistochemical staining. Mean optical density (OD) value of the gene proteins expression were quantitatively examined by image analysis system. Results A.The apoptotic cardiomyocytes were not observed in the sham-operation group. The number of the apoptotic cells were 134.45?45.61/field in the ischemia- reperfusion group, 51.65?13.71/field in the ginsenoside Rb 1-treated group and 90.66? 19.22/field in the ginsenoside Re-treated group, respectively. The differences were significant among the three groups (P0.05), but Bax?Bad and Fas gene expression were decreased significantly in the ginsenoside Rb 1 and Re-treated group as compared with the ischemia-reperfusion group (P

BACKGROUND: The studies found that cardiomyocyte apoptosis is related to ischemia-reperfusion injury directly. The clinical and experimental studies proved that ginseng and losartan could improve myocardial iscbemia and prevent the ischemia-reperfusion injury.But the comparative study of their effects on cardiomyocyte injury induced by ischemia and reperfusion has not been reported.OBJECTIVE: To compare the effects of ginseng and losartan on cardiomyocyte Bcl-2 gene expression after ischemia and reperfusion in vivo.DESIGN: Randomized controlled experiment.SETTING: Department of Cardiovascular Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the Laboratory of Cardiovascular Internal Medicine of Tongji Hospital of Tongji Medical College Affiliated to Huazhong University of Science and Technology from November 2002 to April 2003. Totally 40 healthy adult Wistar rats, weighting 200-250 g, of either gender, were selected and divided into 4 groups:sham operated group, ischemia-reperfusion group, ginseng treated group and losartan treated group with 10 rats in each group.METHODS: Rats were modeled in ischemia-reperfusion group, ginseng treated group and los arran treated group but not in sham operation group.20 mg/kg (1 mL in volume) losartan was given by stomach. The first administration was 2 hours prior to operation. Subsequently, the second and third administrations were given in immediate and 24 hours after operation,respectively; 1 mL of radix ginseng rubra (1 g/mL) was given by stomach.The first administration was 2 hours before operation. Subsequently, the second and third administrations were given in just and 24 hours after operation, respectively. The rats in ischemia-reperfusion and sham-operated group were given the normal saline with the same volume and at the same time. Immunohistochemistry and in situ hybridization histochemistry were used to measure mRNA and protein of Bcl-2 gene expression that were compared with those in the control group.MAIN OUTCOME MEASURES: The expression level of Bcl-2 mRNA and Bcl-2 protein in ginseng treated group and losartan treated group.RESULTS: Data of totally 40 rats was entered the final analysis. ① Content of Bcl-2 mRNA and protein were not significantly different both in ginseng group and control group (P ＞ 0.05). ② Contents of Bcl-2 mRNA and Bcl-2 protein were higher in ginseng treated group than those in the ischemia-reperfusion group (P ＜ 0.05 or 0.01).CONCLUSION: Expressions of Bcl-2 mRNA and Bcl-2 protein in ginseng treated group are higher than those in losartan treated group, which suggests that ginseng has not same effect with losartan in inhibiting cardiomyocyte apoptosis and in preventing and curing cardiomyocyte injury induced by ischemia-reperfusion.

BACKGROUND: Leptin is a kind of polypeptide hormone secreted by fatty tissue, previous studies have shown that leptin plays a certain role in the formation of atherosclerosis.OBJECTIVE: To investigate the effects of leptin on the expression of tumor necrosis factor-alpha (TNF-α) in RAW264.7 cells, and investigate the possible mechanism from the angle of the change of nuclear factor-κB (NF-κB) activity.DESIGN: A controlled observational experiment.SETTING: Department of Biochemistry and Molecular Biology, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiments were carried out in the Department of Biochemistry and Molecular Biology, Tongji Medical College, and the Department of General Surgery, Union Hospital affiliated to Huazhong University of Science and Technology between April 2005 and February 2006.The cultured RAW264.7 cells were divided into leptin treated groups treated with different concentration (12.5, 25, 50, 100 μg/L), I kappa B kinase inhibitor group and blank control group. Each group had 3 sub-wells,and the experiments were repeated for 3 times.METHODS: Mouse macrophage RAW264.7 cells were incubated into a 6-well plate at the density of 109 cells L-1, cultured in RPMI-1640 culture medium containing bovine serum of 0.1 in volume fraction. When the cells grew to 80%, serum-free culture medium Opti-MEM was changed to culture for another 24 hours, and then the cells were divided into leptin groups treated with different concentration (12.5, 25, 50, 100 μg/L), I kappa B kinase inhibitor group and blank control group. After the cells were incubated with leptin for 4 hours, the expression of TNF-α mRNA expression in RAW264.7 cells was detected with reverse transcription-polymerase chain reaction (RT-PCR). After the RAW264.7 cells were incubated with leptin for 1, 3, 6 and 9 hours, the expression of TNF-c protein expression was detected with double antibody sandwich enzyme-linked immunosorbent assay (ELISA). After the RAW264.7 cells were incubated with leptin for different times, the activity of NF-κB was detected analyzed with electrophoretic mobility shift assay. Another, the RAW264.7 cells were treated with or without 50 μmol/L leptin and/or 100 μmol/L PS1145 (I kappa B kinase specific inhibitor)divided into four groups: blank control group, I kappa B kinase specific inhibitor PS1145 (10 μmol/L) treated group, leptin (50 μmol/L) treated group, leptin (50 μmol/L) + PS1145 (10 μmol/L) group. Aftere incubated for 6 hours, the activity of NF-κB and expression TNF-α mRNA were detected respectively.MAIN OUTCOME MEASURES: ① Effect of leptin of different concentration on the expression of TNF-α mRNA and protein in RAW264.7cells; ② Effect of leptin of different concentration on the activity of NF-κB in RAW264.7 cells; ③ Influence of inhibition I kappa B kinase activity inhibition on expression of TNF-a induced by leptin in RAW264.7cells.RESULTS: ① After the RAW264.7 cells were treated with leptin of different concentration, the TNF-α mRNA level was elevated in a dose-dependent manner, and it reached the peak value emerged in the 50 μg/L leptin treated group. ② The expression of TNF-α protein increased in dose-dependent and time-dependent manners, and it reached the peak val ue at 6 hours in the 50 μg/L leptin treated group. ③ The activity of NF-κB was also positively correlated with the leptin concentration, and it was the highest value at 6 hours treated with 50 μg/L leptin (P ＜ 0.05). ④ I kappa B kinase activity inhibition only partially suppressed the leptin induced elevation of TNF-α expression induced by leptin.CONCLUSION: Leptin can increase the expression and secretion of TNF-α in RAW264.7 cells directly in both dose-dependent and time-dependent manners, and the mechanism may be correlated with the activated NF-κB induced by leptin. It may be one of the mechanisms of atherosclerosis induced by leptin.

Objective To investigate the relationship between microalbuminuria to estimated glomerular filtration rate (mALB/GFR) ratio and incidence of contrast induced nephropathy (CIN) after underwent percutaneous coronary intervention (PCI). Between March 2010 to August 2011, a total of 137 patients who underwent PCI were selected. Anthropometric measures, lipid profiles, fasting blood glucose, CRP, base creatinine and microalbuminuria were measured before operation. Glomerular filtration rate was calculated by MDRD formula. Contrast volume (CV) was recorded after the operation for each patient. Serum creatinine was measured 24 h, 48 h, 72 h after operation, respectively. CIN was defined as an increase in serum creatinine levels of 25% or 44 μmol/L or more from baseline within 72 h after PCI. All patients were divided into group A (CIN group) and group B (No CIN group). Results Eighteen (13.1%) patients developed CIN(group A). The others were group B (no CIN group). The level of CRP, base creatinine, microalbuminuria and mALB/GFR in group A were significantly higher than that in group B ( P < 0 . 01 ) . GFR in group A was lower than that in group B (P < 0.05). Logistic regression analysis indicated that the level of base creatinine , microalbuminuria GFR and mALB/GFR were independent risk factors of CIN occurrence (P < 0.05). The receiver-operator characteristic curve analysis indicated that a mALB/ GFR ratio of 1.17 was a fair discriminator for CIN,and sensitivity were 94.1%, specifity were 72.5%. Conclusion A mALB/ GFR ratio was a independent predictor of CIN after PCI.

AIM:Cytochrome P450 epoxygenase 2J2 and epoxyeicosatrienoic acids ( EETs) are known to protect against cardiac hypertrophy and heart failure, which involve activation of 5′-AMP-activated protein kinase ( AMPK) and Akt.Although the functional roles of AMPK and Akt are well established , the significance of crosstalk between them in the development of cardiac hypertrophy and anti -hy-pertrophy of CYP2J2 and EETs remains unclear .Here, we investigated whether CYP 2J2 and its metabolites EETs protected against cardiac hypertrophy by activating AMPKα2 and Akt1.Moreover, we tested whether EETs enhanced crosstalk between AMPKα2 and phosphorylated Akt1 ( p-Akt1), and stimulated the nuclear translocation of p-Akt1, to exert their anti-hypertrophic effects. METHODS:The recombinant rAAV9 vector was coupled to CYP2J2 and the rAAV9-CYP2J2 construct was injected into the caudal vein of AMPKα2-/-and littermate control mice .AMPKα2 -/-and littermate control mice that overexpressed CYP 2J2 in heart were treated with angiotensin II (Ang II) for 2 weeks.Hemodynamic and cardiac functions were also evaluated after 14 days of infusion with Ang II or saline.RESULTS:Interestingly, the overexpression of CYP2J2 suppressed cardiac hypertrophy , including decreased heart size, cross sectional area of cardiomyocytes , markers of cardiac hypertrophy [ brain natriuretic peptide ( BNP) ,β-myosin heavy chain (β-MHC) and skeletal muscle α-actin (ACTA1)] and increased levels of atrial natriuretic peptide (ANP) in the heart tissue and plasma of wild-type mice but not AMPKα2 -/-mice.Measurement of left ventricular ejection fraction and fractional shortening showed that CYP2J2 overexpression prevented Ang II-induced ventricular systolic dysfunction in mice .Moreover, an Ang II-induced reduction in cardiac function, demonstrated by decreased dp/dtmax and dp/dtmin, was prevented by overexpression of CYP2J2.Mechanistically, the CYP2J2 metabolites 11,12-EET activated AMPKα2 to induce the nuclear translocation of p-Akt1, which increased production of ANP and thereby inhibited the development of cardiac hypertrophy .Furthermore , by co-immunoprecipitation analysis , we found that full-length Akt1 and an Akt1 fragment containing amino acids 150-408, which constitute the protein kinase domain , but not other frag-ments of Akt1, bind to the AMPKγ1 subunit.AMPKα2β2γ1 and p-Akt1 interact through the direct binding of the AMPKγ1 subunit to the Akt1 protein kinase domain.This interaction was enhanced by 11,12-EET.CONCLUSION:Our studies reveal a novel mechanism in which CYP2J2 and EETs enhanced Akt1 nuclear translocation through interaction with AMPKα2β2γ1 and protect against cardiac hy-pertrophy and suggest that overexpression of CYP 2J2 might have clinical potential to suppress cardiac hypertrophy and heart failure .

Objective To concentrate on the morbidity of cerebral microbleed (CMB) in patients with hypertension and to analyze the predilection and risk-factor of cerebral microbleed.Method Hypertensive patients were divided into the simple hypertention group, hypertention group with lacunar infarction and hypertention group with cerebral infarction.All of these 65 patients received examination of susceptibility-weighted imaging.Results Ninety-one focuses of cerebral microbleeds were found in these patients:58.2% of these focuses were in both basal ganglia and cerebral ganglion;35.2 percent were in cortex and subcortex;6.6 percent were in brainstem and cerebellum.The total morbidity of CMB was 33.8 percent, 52.4 percent in the group with lacunar infarction and 38.1 percent in the group with cerebral infarction, both were significantly higher than that of 8.7 percent in the simple hypertensive group (χ2= 8.08,P<0.01 andχ2=3.86, P<0.05).Conclusions The focus of CMB suggested the hemorrhagic tendency in endocranial capillary.CMB can be used as a routine exam for the hemorrhagic tendency in endocranial capillary.Synthetic analysis of risk-factor and the result of SWI help clinicians choose suitable treatment for each patient.

Objective To investigate the effect of CD151 gene therapy on improving myocardial function in swines with myocardial infarction. Methods CD151, antisense CD151 and green fluorescent protein (GFP) were constructed into the recombinant adeno-associated virus (rAAV). Swines were divided into 4 groups: rAAV-GFP group (6 swines), rAAV-CD151 group (6 swines), rAAV-antiCD151 group (6 swines) and control group (6 swines). The swines were performed with coronary artery ligation and intramuscularly injection with rAAV. Eight weeks after vector administration, western blot was used to detect gene expression of CD151. 13N-labeled NH3 positron emission tomography (PET) was used to evaluate myocardial perfusion. Echocardiography was used to assess myocardial function. Results Compared with the control group and the rAAV-GFP group, the rAAV-CD151 group showed higher CD151 protein expression. Compared with the rAAV-GFP group, the defect size of myocardium was decreased[( 11.3±2.4)% vs. (21.1±2.6)%, t= -5.67,P<0.01] and left ventricular ejection fraction (EF), left ventricular fractional shortening (FS), the ratio of anterior lateral wall thickening (△ALWT) and ratio of interventricular septum thickening (△IVST) were significantly improved in rAAV-CD151 group 8 weeks after vector administration [(65.7±4.6)% vs. (54.7±5.3)%, (36.0±2.9)% vs. (27.6±3.1)%,(55.4± 4.9)% vs. (36.8±7.8)%, (35.2±6.0)% vs. (26.7±4.4)%, t=3.98, 3.35, 3.34, 9.27, all P< 0.05]. The level of diastolic ALWT and diastolic IVST was also increased in rAAV CD151 group compared with rAAV-GFP group ( P<0.05).Compared with rAAV-CD151 group, parameters of myocardial function in rAAV-antisense CD151 group were not improved (P<0.05). Conclusions rAAV-CD151 can effectively transfeet the myocardium, increase the expression ofCD151 protein, promote the blood perfusion of myocardium and improve the ventricular function after myocardial infarction.

To observe the effects of simvastatin on nuclear factor kappaB (NF-kappaB)-DNA binding activity and on the expression of monocyte chemoattractant protein-1 (MCP-1) in atherosclerotic plaque in rabbits and to explore the anti-atherosclerotic properties beyond its lipid-lowering effects. Thirty-six New Zealand male rabbits were randomly divided into low-cholesterol group (LC), high-cholesterol group (HC), high-cholesterol+simvastatin group (HC+S) and then were fed for 12 weeks. At the end of the experiment, standard enzymatic assays, electrophoretic mobility shiftassay (EMSA), immunohistochemical staining, and morphometry were performed to observe serum lipids, NF-kappaB-DNA binding activity, MCP-1 protein expression, intima thickness and plaque area of aorta respectively in all three groups. Our results showed that the serum lipids, NF-kappaB-DNA binding activity, expression of MCP-1 protein, intima thickness, and plaque area of aorta in the LC and HC+S groups were significantly lower than those in the HC group (P<0.05). There was no significant difference in the serum lipids between the LC and HC+S groups (P>0.05), but the NF-kappaB-DNA binding activity, the expression of MCP-1 protein and the intima thickness and plaque area of aorta in the HC+S group were significantly decreased as compared to the LC group (P<0.05). This study demonstrated that simvastatin could decrease atherosclerosis by inhibiting the NF-kappaB-DNA binding activity and by reducing the expression of MCP-1 protein.

The effects of carvedilol on cardiomyocyte apoptosis and expression of bcl-2, bax genes following ischemia (0.5 h) and reperfusion (48 h) in vivo and the possible biological mechanism of carvedilol inhibiting cardiomyocyte apoptosis were studied. The left anterior descending artery in Wistar rats were ligated to establish ischemia-reperfusion (I/R) models. The model animals were divided into two groups: I/R group, the model rats not subject to other treatments except ischemia-reperfusion (n = 8); carvedilol-treated group (n = 8), I/R model rats treated with carvedilol. Eight rats in the sham-operated group were subjected to only experimental open operation. The number of apoptotic cardiomyocyte was determined by TUNEL staining. Immunohistochemistry and in situ hybridization histochemistry (ISHH) were used to detect the expression of bcl-2 and bax genes. Image processing system was used to quantitatively dispose the positive metric substances of both immunohistochemistry and ISHH through the average optical density (OD) value. The results showed that the number of the apoptotic cells were 36.18 +/- 9 (I/R group), 0-1 (sham-operated group) and 9.5 +/- 3 (carvedilol-treated group) in each visual field respectively with the difference being very significant among the groups (P < 0.001). The OD values of bcl-2 protein in sham-operated group, I/R group and carvedilol-treated group were 0.14 +/- 0.01, 0.08 +/- 0.02 and 0.15 +/- 0.03, respectively. The OD values of bcl-2 mRNA in sham-operated group, I/R group and carvedilol-treated group were 0.08 +/- 0.01, 0.06 +/- 0.01 and 0.09 +/- 0.01, respectively. There was no significant difference between carvedilol-treated group and I/R group (P > 0.05). The OD values of bax protein in I/R group, sham-operated and carvedilol-treated-treated group were 0.13 +/- 0.02, 0.07 +/- 0.01, 0.06 +/- 0.01, respectively. There was very significant difference between carvedilol-treated group and I/R group (P < 0.01). Bcl-2/bax ratio was 1.07 +/- 0.14 (I/R group), 1.28 +/- 0.16 (sham-operated group), 2.5 +/- 0.26 (carvedilol-treated group) respectively with the difference being very significant between carvedilol-treated group and I/R group (P < 0.05). It was indicated that carvedilol could inhibit cardiomyocyte apoptosis following ischemia and reperfusion, which was related to the increased bcl-2/bax ratio due to inhibition of bax gene expression.
Adrenergic alpha-Antagonists ; pharmacology ; Adrenergic beta-Antagonists ; pharmacology ; Animals ; Apoptosis ; drug effects ; Carbazoles ; pharmacology ; Gene Expression ; Myocardial Ischemia ; genetics ; pathology ; Myocardial Reperfusion Injury ; genetics ; pathology ; Myocytes, Cardiac ; pathology ; Propanolamines ; pharmacology ; Proto-Oncogene Proteins ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; bcl-2-Associated X Protein

To observe the effects of simvastatin on nuclear factor kappaB (NF-κB)-DNA binding activity and on the expression of monocyte chemoattractant protein-1 (MCP-1) in atherosclerotic plaque in rabbits and to explore the anti-atherosclerotic properties beyond its lipid-lowering effects.Thirty-six New Zealand male rabbits were randomly divided into low-cholesterol group (LC), highcholesterol group (HC), high-cholesterol+ simvastatin group (HC+S) and then were fed for 12weeks. At the end of theexperiment, standard enzymatic assays, electrophoretic mobility shift assay (EMSA), immunohistochemical staining, and morphometry were performed to observe serum lipids, NF-κB-DNA binding activity, MCP-1 protein expression, intima thickness and plaque area of aorta respectively in all three groups. Our results showed that the serum lipids, NF-κB-DNA binding activity, expression of MCP-1 protein, intima thickness, and plaque area of aorta in the LC and HC+S groups were significantly lower than those in the HC group (P＜0.05). There was no significant difference in the serum lipids between the LC and HC+S groups (P＞0.05), but the NF-κB-DNA binding activity, the expression of MCP-1 protein and the intima thickness and plaque area of aorta in the HC+S group were significantly decreased as compared to the LC group (P＜0.05). This study demonstrated that simvastatin could decrease atherosclerosis by inhibiting the NFκB-DNA binding activity and by reducing the expression of MCP-1 protein.