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Objective:To evaluate the role of ferroptosis in hypoxia-reoxygenation (H/R) injury in cardiomyocytes cultured in high-fat high-glucose (HFHG) medium.Methods:Cardiomyocytes H9c2 cells were commonly cultured and divided into 3 groups ( n=20 each) using a random number table method: control group (C group), HFHG-H/R group and Ferrostatin-1 (Fer-1) plus HFHG-H/R group (Fer-1+ HFHG+ H/R group). H9c2 cells were cultured in a HFHG medium for 12 h and then exposed to 1%O 2-5%CO 2-94%N 2 for 4 h, followed by 2 h reoxygenation in a cell incubator.Fer-1 at a final concentration of 10 μmol/L was added while the cells were cultured in the HFHG medium in group Fer-1+ HFHG+ H/R.At 2 h of reoxygenation, the cell viability was measured using CCK-8 assay, the activity of lactate dehydrogenase (LDH) in the supernatant was measured using 2, 4-dinitrophenylhydrazine color method, the activity of reactive oxygen species (ROS) was measured by fluorescent probe DCFH-DA flow cytometry, and the expression of acyl-CoA synthetase long-chain family member 4 (ACSL4), nuclear receptor coactivator 4 (NCOA4), and glutathione peroxidase 4 (GPX4) was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the activities of LDH release and ROS were increased, and the expression of ACSL4 and NCOA4 was up-regulated ( P<0.05), and no significant change was found in the expression of GPX4 in group HFHG+ H/R ( P>0.05). Compared with group HFHG+ H/R, the cell activity was significantly increased, the activities of LDH and ROS were decreased, and the expression of ACSL4 and NCOA4 was down-regulated ( P<0.05), and no significant change was found in the expression of GPX4 in Fer-1+ HFHG+ H/R group ( P>0.05). Conclusions:Ferroptosis is involved in the process of H/R injury in cardiomyocytes cultured in HFHG medium.
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Objective:To evaluate the role of hypoxia-inducible factor-1α (HIF-1α) in the renal injury induced by myocardial ischemia-reperfusion (I/R) in diabetic rats and its relationship with solute carrier family7 member11 (SLC7A11).Methods:SPF-grade healthy male Sprague-Dawley rats, aged 4 weeks, weighing 100-130 g, were fed with high-fat and high-sucrose diet freely.The weight of the rats was measured once a week.After the weight of the animals reached 240 g, 1% streptozotocin (STZ)-citrate buffer 35 mg/kg was injected intraperitoneally to induce type 2 diabetes mellitus.After injection of STZ, the animals were fed with high-fat and high-sucrose diet continuously.Blood samples were collected from the tail vein for determination of blood glucose concentrations 1 week later.When random blood glucose was ≥16.7 mmol/L for 3 times, the model of type 2 diabetes mellitus was considered to be established successfully.After the model was established successfully, the animals were fed with high-fat and high-sucrose diet continuously for 6 weeks.Eighteen rats with type 2 diabetes mellitus were selected and divided into 3 groups ( n=6 each) using a random number table method: diabetic sham operation group (group DS), diabetic myocardial I/R group (group DIR) and diabetic myocardial I/R+ HIF-1α agonist DMOG group (DIR+ DMOG group). Twelve non-diabetic rats were divided into 2 groups ( n=6 each) using a random number table method: non-diabetic sham operation group (NS group) and non-diabetic myocardial I/R group (NIR group). The rat myocardial I/R injury model was established by ligating the anterior descending branch of the left coronary artery for 30 min followed by 120 min reperfusion in anesthetized rats.Blood samples were collected from the right internal carotid artery at 120 min of reperfusion for determination of the serum creatinine (Cr), urea nitrogen (BUN) and neutrophil gelatinase-associated lipocalin (NGAL) concentrations (by enzyme-linked immunosorbent assay). Renal tissues were obtained for examination of the pathological changes (by HE staining method) and for determination of the expression of HIF-1α and SLC7A11 (by Western blot). The damage to the renal tubules was scored. Results:Compared with group NS, the concentrations of serum Cr, BUN and NGAL and renal tubular damage score were significantly increased in group DS and group NIR, the expression of HIF-1α and SLC7A11 was down-regulated in group DS, and the expression of HIF-1α and SLC7A11 was up-regulated in group NIR ( P<0.05). Compared with group DS, the concentrations of serum Cr, BUN and NGAL and renal tubular damage score were significantly increased, and the expression of HIF-1α and SLC7A11 was up-regulated in group DIR ( P<0.05). Compared with group NIR, the concentrations of serum Cr, BUN and NGAL and renal tubular damage score were significantly increased, and the expression of HIF-1α and SLC7A11 was down-regulated in group DIR ( P<0.05). Compared with group DIR, the concentrations of serum Cr, BUN and NGAL and renal tubular damage score were significantly decreased, and the expression of HIF-1α and SLC7A11 was up-regulated in group DIR+ DMOG ( P<0.05). Conclusion:HIF-1α is involved in the renal injury induced by myocardial I/R, which is related to regulation of the expression of SLC7A11 in rats.
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Objective:To evaluate the role of Notch1/hairy and enhancer of split homolog1(Hes1) signaling pathway in high glucose and hypoxia-reoxygenation (H/R) injury to cardiomyocytes.Methods:H9c2 cardiomyocytes were cultured in low-glucose DMEM culture medium supplemented with 10% fetal bovine serum.The cells were divided into 6 groups ( n=12 each) using a random number table method: control group (group C), H/R group, H/R+ Jagged-1 group (group H/R+ J), high glucose group (group HG), high glucose+ H/R group (group HG+ H/R) and high glucose+ H/R+ Jagged-1 group (group HG+ H/R+ J). The cells were incubated in low-glucose culture medium for 72 h in group C. After incubated in low-glucose culture medium for 72 h, the cells were exposed to 24-h hypoxia in an incubator filled with 95% N 2-5% CO 2 at 37℃, immediately followed by 6-h reoxygenation in an incubator filled with 95% O 2-5% CO 2 at 37℃ in group H/R.In group H/R+ J, Jagged-1 (Notch1/Hes1 signaling pathway specific activator) 5μg/ml was added to low-glucose culture medium and the cells were incubated for 72h before H/R.In group HG, H9c2 cardiomyocytes were incubated in high-glucose culture medium containing 33 mmol/L glucose for 72 h. In group HG+ H/R, the cells were incubated in high-glucose medium for 72 h before H/R.In group HG+ H/R+ J, Jagged-1 5μg/ml was added to high-glucose culture medium, and the cells were incubated for 72 h before H/R.At 6 h of reoxygenation, the supernatant of the culture medium was collected for detection of the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH), the cell viability (by CCK-8 assay) and the cell apoptosis rate (by flow cytometry) and for determination of expression of Notch1, Hes1 and c-caspase-3 (by Western blot). Results:Compared with group C, the cell survival rate and SOD activity were significantly decreased, and apoptosis rate and LDH activity were increased in H/R, H/R+ J and HG groups, expression of Notch1, Hes1 and c-caspase-3 was up-regulated in H/R and H/R+ J groups, and the expression of Notch1 and Hes1 was down-regulated and c-caspase-3 expression was up-regulated in group HG ( P<0.05). Compared with group H/R, the cell survival rate and SOD activity was significantly increased, apoptosis rate and LDH activity were decreased, expression of Notch1 and Hes1 was up-regulated, and c-caspase-3 expression was down-regulated in group H/R+ J, and the cell survival rate and SOD activity were significantly decreased, apoptosis rate and LDH activity were increased, expression of Notch1 and Hes1 was down-regulated, and c-caspase-3 expression was up-regulated in group HG+ H/R ( P<0.05). Compared with group HG, the cell survival rate and SOD activity were significantly decreased, and apoptosis rate and LDH activity were increased in HG+ H/R and HG+ H/R+ J groups ( P<0.05), and expression of Notch1 and Hes1 was down-regulated, and c-caspase-3 expression was up-regulated in group HG+ H/R ( P<0.05). Compared with group HG+ H/R, the cell survival rate and SOD activity were significantly increased, apoptosis rate and LDH activity were decreased, expression of Notch1 and Hes1 was up-regulated, and c-caspase-3 expression was down-regulated in group HG+ H/R+ J ( P<0.05). Compared with group H/R+ J, the cell survival rate and SOD activity were significantly decreased, apoptosis rate and LDH activity were increased, expression of Notch1 and Hes1 was down-regulated, and c-caspase-3 expression was up-regulated in group HG+ H/R+ J ( P<0.05). Conclusion:Activation of Notch1/Hes1 signaling pathway is the endogenous protective mechanism of high glucose and H/R injury to cardiomyocytes.
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Objective:To observe influence of alprostadil injection on contrast-induced nephropathy (CIN) in high risk patients after percutaneous coronary intervention (PCI).Methods: A total of 263 CIN high risk (CIN risk score ≥16 scores) patients were selected.According to random number table, patients were randomly divided into routine treatment group (n=121, received routine hydration therapy) and alprostadil group (n=142, received additional alprostadil injection based on routine treatment group).Serum creatinine (SCr), glomerular filtration rate(GFR), cystatin C (CysC) and β trace protein (β-TP) level before, 48h and 72h after PCI were measured and compared, and incidence rate of CIN, percentage of blood purification therapy and mortality were compared between two groups.Results: Compared with before PCI, there was significant rise in SCr level and significant reduction in GFR in both groups on 48h and 72h after PCI (P<0.01 all);Compared with routine treatment group, there were significant reductions in levels of SCr [72h: (190.04±28.92) μmol/L vs.(141.10±21.18) μmol/L], and significant rise in GFR [72h: (26.0±4.4) ml/min vs.(36.4±4.9) ml/min], and levels of CysC[72h: (1.75±0.74) mg/L vs.(1.47±0.55) mg/L] and β-TP [72h: (1.53±0.50) mg/L vs.(1.22±0.38) mg/L] significantly decreased in alprostadil group on 48h and 72h after PCI, P<0.05 or <0.01;there were significant reductions in incidence rate of CIN (30.6% vs.18.3%) and percentage of blood purification therapy (10.7% vs.3.5%) in alprostadil group, P=0.001, 0.045 respectively.There was no significant difference in mortality between two groups, P=0.728.Conclusion: Alprostadil injection can significantly improve kidney function, reduce incidence rate of CIN and percentage of blood purification therapy in CIN high risk patients after PCI, which is worth extending.
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Weanling rats and mice were fed on an iron-deficient diet. By the end of the 4th week, the rats were sacrificed for the determination of immune function and the mice were infected with S. typhimurium. The rate of peripheral blood lymphocytes transformation, total serum complement activity (CH50) and splenic tissue cAMP levels in the iron-deficient rats were 26.3? 5.86%, 58.39?14.79u/ml and 0.93?0.48 pmol/mg respectively, but that in the controls were 38.3?4.61%, 75.17?19.51u/ml and 2.69?1.17pmol/mg respectively. The former were significantly lower than the latter. The serum hemolysin (HC50) and serum lysozyme in the iron-deficient rats were 330.62 ?21.90 and 29.50 ?16.85ug/ml respectively; and that in the controls were 353.24?29.77 and 30.08?10.39ug/ml respectively. The differences between these two groups were not significant. Gross histological changes were found in the spleen of the iron-deficient rats, but no changes in the thymus. After the intraperitoneal injection of S. typhimurium, the mortality of iron-deficient mice were significantly higher than the controls.