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Objective: To observe the effects of acute exhaustive exercise on the expressions of oxidative stress related enzymes in skeletal muscle of rats.
Methods: Forty male SD rats were divided into 4 groups, 10 rats in each group, which were the control group (C group), exhausted exercise group (E group), exercise + PKC inhibitor group (EC group), exercise + NOX inhibitor group (EA group). Three groups of exercise rats were familiarized with treadmill running for 3 days (5 m/min, once/d, no incline), then rested for one day. EC group was injected with PKC inhibitor chelerythrine (5 mg / kg) one day before and one hour before exercise, EA group was injected with NADPH oxidase inhibitor apocynin (10 mg / kg) at the same time, group C and group E were injected with the same dose of normal saline. Three groups of exercise rats were subjected to a one-time treadmill exhaustion exercise, and the plantaris were taken after exhaustion. Reactive oxygen species (ROS) were detected by DCF fluorescent probe, NOX2, NOX4, 3-NT were analyzed by Western blot, and PKC, NOX2, NOX4 were analyzed by immunoprecipitation.
Results: Compared with group C, ROS level, NOX2 and NOX4 protein expressions, PKC-NOX2 and PKC-NOX4 complex levels, and 3-NT production in group E were significantly increased (P＜0.01, P＜0.05), and ROS level was no significant difference in group EC and group EA (P＞0.05), and NOX4 protein expression in group EC was significantly increased (P ＜ 0.05). Compared with group E, ROS level, NOX2 and NOX4 protein expressions, PKC-NOX2 and PKC-NOX4 complex levels, 3-NT production were decreased significantly (P＜0.01, P＜0.05).
Conclusion: Exhausted exercise induces increased expressions of NOX2 and NOX4 proteins in skeletal muscle, and PKC mediates the production of ROS by regulating NOX2.
[Mechanism of oxidative stress in skeletal muscle of rats induced by acute exhaustive exercise]
Jiao Liu 1, Gang Zhou 1, Yu Mei 1, Wen-Jie Xie 1, Peng-Fei Li 1, Fan Yang 1
2020 Jan 28;
Aims: Vascular dysfunction plays a key role in sepsis but the role of perivascular adipose tissue (PVAT) in this condition is relatively unknown.
Main methods: Sepsis was induced by cecal ligation and puncture (CLP). The responses of the aorta and superior mesenteric artery to norepinephrine in the presence or absence of PVAT were evaluated. Fluorescent probes measured the production of nitric oxide (NO) and reactive oxygen species (ROS). NO synthases (NOS) and β3-adrenoceptor expression were detected by immunofluorescence and S-nitrosylation by the biotin switch assay.
Key findings: Aorta and superior mesenteric arteries from septic animals with intact PVAT showed a worsened response to the vasoconstrictor compared to vessels without PVAT. PVAT from the aorta (APVAT) produced NO and ROS whereas PVAT from the superior mesenteric artery (MPVAT) produced only ROS. Septic APVAT exhibited a higher density of NOS-1 and NOS-3. S-nitrosylation was found in APVAT. Donor (PVAT obtained from normal or septic rats):Host (normal vessel without PVAT) experiments showed that L-NAME, ODQ and β3-adrenergic receptor antagonist blocked the septic APVAT anti-contractile effect. None of these compounds affected MPVAT; tempol, but not apocynin, blocked its anti-contractile effect.
Significance: PVAT contributes to the anti-contractile effect in the aorta and mesenteric artery of septic rats through different pathways. β3-Adrenergic receptor and NO appear to be key mediators of this effect in APVAT, but not in MPVAT where ROS seem to be a relevant mediator. Therefore, PVAT is a relevant player of sepsis vascular dysfunction.
Nitric oxide; Perivascular adipose tissue; Reactive oxygen species; Sepsis; Vascular dysfunction.
Perivascular adipose tissue phenotype and sepsis vascular dysfunction: Differential contribution of NO, ROS and beta 3-adrenergic receptor
Clarissa Germano Barp 1, Patricia Oliveira Benedet 1, Jamil Assreuy 2
2020 Aug 15
Renal ischemia/reperfusion injury is a common cause of acute kidney injury (AKI) and hypertension might contribute to the increased incidence of AKI. The purpose of this study was to investigate the effects of single and combined hyperbaric oxygen (HBO) preconditioning and NADPH oxidase inhibition on oxidative stress, kidney function and structure in spontaneously hypertensive rats (SHR) after renal ischemia reperfusion injury. HBO preconditioning was performed by exposing to pure oxygen (2.026 bar) twice a day for two consecutive days for 60 minutes, and 24h before AKI induction. For AKI induction, the right kidney was removed and ischemia was performed by clamping the left renal artery for 45 minutes. NADPH oxidase inhibition was induced by apocynin (40 mg/kg b.m., intravenously) 5 minutes before reperfusion. AKI significantly increased renal vascular resistance and reduced renal blood flow, which were significantly improved after apocynin treatment. Also, HBO preconditioning, with or without apocynin treatment showed improvement on renal hemodynamics. AKI significantly increased plasma creatinine, urea, phosphate levels and lipid peroxidation in plasma. Remarkable improvement, with decrease in creatinine, urea and phosphate levels was observed in all treated groups. HBO preconditioning, solitary or with apocynin treatment decreased lipid peroxidation in plasma caused by AKI induction. Also, combined with apocynin, it increased catalase activity and solitary, glutathione reductase enzyme activity in erythrocytes. While AKI induction significantly increased plasma KIM- 1 levels, HBO preconditioning, solitary or with apocynin decreased its levels. Considering renal morphology, significant morphological alterations present after AKI induction were significantly improved in all treated groups with reduced tubular dilatation, tubular necrosis in the cortico-medullary zone and PAS positive cast formation. Our results reveal that NADPH oxidase inhibition and hyperbaric oxygen preconditioning, with or without NADPH oxidase inhibition may have beneficial effects, but their protective role should be evaluated in further studies.
Hyperbaric oxygen preconditioning and the role of NADPH oxidase inhibition in postischemic acute kidney injury induced in spontaneously hypertensive rats
Sanjin Kovacevic 1, Milan Ivanov 2, Zoran Miloradovic 2, Predrag Brkic 3, Una Jovana Vajic 2, Maja Zivotic 4, Nevena Mihailovic-Stanojevic 2, Djurdjica Jovovic 2, Danijela Karanovic 2, Rada Jeremic 3, Jelena Nesovic-Ostojic 1
2020 Jan 8