Reactive oxygen species (ROS) play an important role in the pathogenesis of various cardiovascular diseases, by leading to cell apoptosis and thus causing organic injuries. Anti-ROS therapy is highly anticipated, but currently, there is still no appropriate prevention method. Studies have shown that thioredoxin (Trx), being a kind of significant endogenous antioxidant system, has excellent antioxidant capacity. Promotion of Trx can reduce key biomolecules to eliminate ROS or regulate many signaling pathways, thus resisting ROS injuries, which may be a new anti-ROS strategy. Therefore, we reviewed the research progress of Trx in cardiac antioxidant therapy to discuss its potential and possibility to be a target for prevention of heart-related ROS injury.
The aim of this research is to investigate the influence of microencapsulation on the expression of the oxidative stress genes and exogenous regulation of HepG2 cells. We compared the expression of hemeoxygenase-1 (HO-1) and glutathione S-transferases-A1 (GST-A1) in HepG2 cells under different culture conditions through real-time PCR. The effects of exogenous antioxidants on cell viability and albumin levels were also evaluated through MTT assay and ELISA assay. The results showed that after culturing for 6 and 16 days, the expression levels of HO-1 in encapsulated cells were approximately 4.9 and 3.1 times higher than that of monolayer cells at the same culture period; As for the expression levels of GST-A1, they were elevated to 11.2 and 33 times of monolayer cells (P<0.05). Accordingly, we found that NAC at 5-10 mmol/L significantly increased the viability by 40%-70% and the biosynthetic function by 20%-30% in microencapsulated HepG2 cells (P<0.05). GSH increased the viability of the encapsulated cells by 20%-55% and the biosynthetic function by 15% (P<0.05). In conclusion, oxidative stress exists in the microcapsules and affects genes expression. Exogenous antioxidants can prevent the inhibition effects of oxidative stress on cellular growth.
Objective To investigate the efficacy of phloretin combined with sodium hyaluronate in preventing postoperative abdominal adhesion formation in rats and its possible mechanisms. Methods Forty rats were randomly divided into five groups, the rats in the sham-operatinon group only underwent open and closed abdominal surgery, and the remaining rats of four groups underwent cecum scratch-and-rub method of modeling to receive different treatments: the rats in the control group and the phloretin group (PHL group) were closed abdominally after modeling, while the rats in the sodium hyaluronate group (HA group) and the phloretin combined with sodium hyaluronate group (PHL+HA group) were closed abdominally by using 2 mL of sodium hyaluronate gel coated with the damaged abdominal wall and the cecum; the postoperative groups treated with phloretin (the PHL and PHL+HA groups) were treated with 2 mL of40 mg/kg phloretin dissolved in 0.5% sodium carboxymethylcellulose by gavage daily, and the rest of the groups were treated with 2 mL of 0.5% sodium carboxymethylcellulose solution by gavage. After general anesthesia, the rats were executed on the 7th day after surgery, and the Nair’s score was used to evaluate the adhesion status of each group on the 7th day after surgery; the adhesive tissue or normal peritoneal tissue were collected (cecum and its opposite side of the peritoneal tissue was collected in the sham-operation group), and immunohistochemistry was performed to evaluate the degree of staining with Nrf2 antibody, HE staining was performed to evaluate the inflammation scores, and Sirius red staining was performed to evaluate the thickness of the collagen fibers, and levels of transforming growth factor β1 (TGF-β1), malondialdehyde (MDA) and superoxide dismutase (SOD) were measured. Results All rats successfully completed the experiment. Compared with the control group, Nair’s score, inflammation score, expression level of TGF-β1, thickness of collagen fibers in the adherent tissues, and MDA level were significantly lower in the PHL+HA group (P<0.05), but the SOD level and expression lever of Nrf2 were significantly higher in the PHL+HA group (P<0.05). Conclusion Phloretin combined with sodium hyaluronate can prevent the formation of postoperative abdominal adhesions in the rat model, which may be related to reducing inflammation, reducing collagen deposition, activating Nrf2 pathway and inhibiting oxidative stress.
ObjectiveTo investigate the effect of nicotinamide mononucleotide adenosyl transferase 3 (NMNAT3) on the mitochondrial function and anti-oxidative stress of rabbit bone marrow mesenchymal stem cells (BMSCs) under oxidative stress in vitro by regulating nicotinamide adenine dinucleotide (NAD+) levels.MethodsThe bone marrow of femur and tibia of New Zealand white rabbits were extracted. BMSCs were isolated and cultured in vitro by density gradient centrifugation combined with adherent culture. The third generation cells were identified by flow cytometry and multi-directional induction. Overexpression of NMNAT3 gene was transfected into rabbit BMSCs by enhanced green fluorescent protein (EGFP) labeled lentivirus (BMSCs/Lv-NMNAT3-EGFP), and then the expression of NMNAT3 was detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot and cell proliferation by cell counting kit 8 (CCK-8) method. BMSCs transfected with negative lentivirus (BMSCs/Lv-EGFP) and untransfected BMSCs were used as controls. The oxidative stress injury cell model was established by using H2O2 to treat rabbit BMSCs. According to the experimental treatment conditions, they were divided into 4 groups: Group A was normal BMSCs without H2O2 treatment; untransfected BMSCs, BMSCs/Lv-EGFP, and BMSCs/Lv-NMNAT3-EGFP in groups B, C, and D were treated with H2O2 simulated oxidative stress, respectively. The effects of NMNAT3 on the mitochondrial function of BMSCs under oxidative stress [changes of mitochondrial membrane potential, NAD+ and adenosine triphosphate (ATP) levels], the changes of anti-oxidative stress ability of BMSCs [reactive oxygen species (ROS) and malondialdehyde (MDA) levels, manganese superoxide dismutase (Mn-SOD) and catalase (CAT) activities], and the effects of BMSCs on senescence and apoptosis [senescence associated-β-galactosidase (SA-β-gal) staining and TUNEL staining] were detected after 24 hours of treatment.ResultsThe rabbit BMSCs were successfully isolated and cultured in vitro. The stable strain of rabbit BMSCs with high expression of NMNAT3 gene was successfully obtained by lentiviral transfection, and the expressions of NMNAT3 gene and protein significantly increased (P<0.05). There was no significant difference in the trend of cell proliferation compared with normal BMSCs. After treatment with H2O2, the function of mitochondria was damaged and apoptosis increased in all groups. However, compared with groups B and C, the group D showed that the mitochondrial function of BMSCs improved, the membrane potential increased, the level of NAD+ and ATP synthesis of mitochondria increased; the anti-oxidative stress ability of BMSCs enhanced, the levels of ROS and MDA decreased, and the activities of antioxidant enzymes (Mn-SOD, CAT) increased; and the proportion of SA-β-gal positive cells and the rate of apoptosis decreased. The differences in all indicators between group D and groups B and C were significant (P<0.05).ConclusionNMNAT3 can effectively improve the mitochondrial function of rabbit BMSCs via increasing the NAD+ levels, and enhance its anti-oxidative stress and improve the survival of BMSCs under oxidative stress conditions.
Vascular cognitive impairment (VCI), a syndrome induced by cerebrovascular disease and its risk factors, has become a major public health challenge worldwide. Especially in the context of an increasingly aging population, its impact is becoming more significant. In recent years, research has gradually revealed the crucial role of chronic cerebral hypoperfusion (CCH) in the occurrence and development of VCI. CCH leads to long-term ischemia and hypoxia in brain tissue, which seriously threatens mitochondrial function and triggers a series of problems such as mitochondrial oxidative stress, calcium homeostasis disturbance, dynamic abnormalities, autophagy dysregulation, and impaired biogenesis. These issues are extensively involved in the pathological process of VCI. This article provides an overview of the correlation between mitochondrial dysfunction and VCI under CCH conditions, aiming to explore new directions for the treatment of VCI.
Acute lung injury (ALI), in which various factors inside and outside the lung lead to hypoxemic respiratory insufficiency and even the development of acute respiratory distress syndrome, has a high morbidity and mortality rate, and its pathogenesis is characterized by complex signaling pathways and limited therapeutic options. A large number of studies have reported that nuclear factor kappa B (NF-κB), phosphatidylinositol 3-kinase (PI3K)/Akt, mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK), vascular endothelial growth factors (VEGF) and JAK/signal transducer and activator of transcription (STAT) signaling pathways are all related to the inflammatory response of ALI, and they are involved in regulating the inflammatory response process of ALI individually or cooperatively. Therefore, this article reviews the research progress on the pathogenesis-related signaling pathways and the drug interventions, aiming to provide a reference for early intervention in lung injury, optimizing the donor pool to increase the proportion of donation after cardiac death and providing quality donor protection conditions.
Objective To investigate the effect of S-adenosyl-l-methionine (SAM) on oxidative stress and alveolar septal cell apoptosis in mice with emphysema after smoking cessation. Methods Twenty-two male SPF C57BL/6J mice aged 6 - 8 weeks were randomly divided into 4 groups, ie. a healthy control group, an emphysema group, a smoking cessation group, and a SAM intervention for 8 weeks after smoking cessation group, with 8 mice in each group. The mice model of emphysema was established by intraperitoneal injection of cigarette smoke extract (CSE) combined with cigarette smoke exposure. Smoking cessation started after the emphysema model was successfully constructed and lasted for 8 weeks. After smoking cessation, the mice in SAM intervention groups were intraperitoneally injected with SAM mg·kg–1·d–1 for 8 weeks. The right lung sections of the mice were taken for hematoxylin-eosin staining to observe pathological changes, and the mean linea rintercept (MLI) and mean alveola rnumber (MAN) of lungs were measured. The concentrations of malondialdehyde (MDA), superoxide-dismutase (SOD) and glutathione (GSH) in alveolar lavage fluid of left lung were detected by spectrophotometry. Terminal deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL) technique was carried out to detect the alveolar septal cells apoptosis. Results MLI, apoptosis index of alveolar septal cell and MDA concentration in bronchoalveolar lavage fluid (BALF) increased significantly in the emphysema group compared with healthy controls, increased significantly in the smoking cessation group compared with the emphysema group, and decreased in the SAM intervention group compared with the smoking cessation group (all P<0.05). GSH concentration and SOD activity in BALF and MAN was significantly lower in the emphysema group compared with the healthy control group, significantly lower in the smoking cessation group compared with the emphysema group, and significantly higher in the SAM intervention group compared with the smoking cessation group (all P<0.05). Conclusions Lung histopathology and apoptosis of alveolar septal cells in emphysema mice progress continuously after smoking cessation. SAM may reduce oxidative stress and improve apoptosis of alveolar septal cells, so as to protect emphysema mice after smoking cessation.
ObjectiveTo investigate the effect of ozone on oxidative stress and energy metabolism change of blood from aortic dissection (AD) patients for providing preliminary evidence of application of ozonated autohemotherapy (ozone-AHT) in AD patients. MethodsTwenty AD patients (16 males and 4 females with a mean age of 48.51±10.21 years) were consecutively included in the First Affiliated Hospital of Harbin Medical University from March 2016 to August 2016, and blood samples were collected from all participants and ozonized in vitro at different ozone concentrations (0 μg/ml, 40 μg/ml, 60 μg/ml, 80 μg/ml, 160 μg/ml). Malondialdehyde (MDA), red blood cells (RBCs) superoxide dismutase (SOD), Na+-K+-ATP, 2,3-bisphosphoglyceric acid (2,3-DPG) at different ozone concentrations were evaluated by enzyme-linked immunosorbent assay (ELISA). ResultsIn the control group (0 μg/ml), the content of postoperative MDA was significantly higher than that of preoperation (P<0.05). The contents of postoperative SOD, Na+-K+-ATP and 2,3-DPG were significantly lower than that of preoperation (P<0.05). The content of MDA at the concentrations of 40 μg/ml, 60 μg/ml, 80 μg/ml group increased after the operation (P>0.05), and the SOD, Na+-K+-ATP, 2,3-DPG decreased compared with the preoperation (P>0.05). But all the values were not statistically significant at the concentrations of 40 μg/ml, 80 μg/ml and 160 μg/ml respectively between preoperation and postoperation (P>0.05). Compared with other concentration groups, the content of preoperative and postoperative MDA increased in the ozone group (160 μg/ml), and oppositely, the contents ofpreoperative and postoperative SOD, Na+-K+- ATP and 2,3-DPG decreased (P<0.05). Conclusion The concentrations of 40 to 80 μg/ml of ozone can improve the antioxidant capacity of erythrocyte membrane, reduce oxidative stress in blood samples of AD patients and improve the energy metabolism of erythrocyte membranes, so the concentration range of ozone is safe and feasible for the ozone-AHT of perioperative AD.
ObjectiveTo explore the effect and mechanism of miR-21 down-regulated which was induced by H2O2 on osteogenic differentiation of MC3T3-E1 cells.MethodsMC3T3-E1 cells were cultured and passaged, and the 7th generation cells were harvested to use in experiment. The MC3T3-E1 cells were treated with different concentrations (0, 40, 80, 160, and 320 μmol/L) of H2O2. The expression of miR-21 was detected by real-time quantitative PCR (RT-PCR) and the cell viability was determined by MTS. Then the appropriate concentration of H2O2 was obtained. To analyze the effect of H2O2 on osteogenic differentiation of MC3T3-E1 cells, the MC3T3-E1 cells were divided into blank control group (group A), H2O2 group (group B), osteogenic induction group (group C), and H2O2+osteogenic induction group (group D). The expression of miR-21 and the osteogenesis related genes expressions of Runx2, osteopontin (OPN), and collagen type Ⅰ alpha 1 (Col1a1) were detected by RT-PCR. The expression of phosphatase and tensin homolog (PTEN) was detected by Western blot. The extracellular calcium deposition was detected by alizarin red staining. To analyze the effect on osteogenic differentiation of MC3T3-E1 cells after the transfection of miR-21 inhibitor and siRNA-PTEN, the MC3T3-E1 cells were divided into H2O2 group (group A1), H2O2+osteogenic induction group (group B1), H2O2+osteogenic induction+miR-21 inhibitor group (group C1), and H2O2+osteogenic induction+miR-21 inhibitor negative control group (group D1); and H2O2 group (group A2), H2O2+osteogenic induction group (group B2), H2O2+osteogenic induction+siRNA-PTEN negative control group (group C2), and H2O2+osteogenic induction+siRNA-PTEN group (group D2). The osteogenesis related genes were detected by RT-PCR and the extracellular calcium deposition was detected by alizarin red staining.ResultsThe results of MTS and RT-PCR showed that the appropriate concentration of H2O2 was 160 μmol/L. The expression of miR-21 was significantly lower in group B than in group A at 1 and 2 weeks (P<0.05). The expression of miR-21 was significantly lower in group D than in group C at 1 and 2 weeks (P<0.05). The expression of PTEN protein was significantly lower in group C than in groups A and D (P<0.05). The mRNA expressions of Runx2, OPN, and Col1a1 were significantly lower in group D than in group C at 1 and 2 weeks (P<0.05). The extracellular calcium deposition in group D was obviously less than that in group C. The expression of PTEN protein was significantly higher in group C1 than in group D1 (P<0.05). The mRNA expressions of Runx2 and OPN were significantly lower in group C1 than in groups B1 and D1 at 1 and 2 weeks (P<0.05). The mRNA expression of Col1a1 was significantly lower in group C1 than in groups B1 and D1 at 2 weeks (P<0.05). The extracellular calcium deposition in group C1 was obviously less than those in groups B1 and D1. The mRNA expressions of OPN and Col1a1 were significantly higher in group D2 than in groups B2 and C2 at 1 week (P<0.05). The extracellular calcium deposition in group D2 was obviously more than those in groups B2 and C2.ConclusionH2O2 inhibits the osteogenic differentiation of MC3T3-E1 cells, which may be induced by down-regulating the expression of miR-21.
Objective To explore the effect of total glucosides of Cistanche deserticola on oxidative stress and cognitive function in rats with intermittent hypoxia. Methods Adult male Wistar rats (n=72) were randomly divided into three groups: a blank control group, a 5% intermittent hypoxia group (IH group) and a total glucosides of Cistanche deserticola intervention group (TGs intervention group). The 5% intermittent hypoxia rat model was simulated by using the self-made cabin of intermittent hypoxia. The rats in the IH group and the TGs intervention group were given 5% intermittent hypoxia respectively, and the rats in the TGs intervention group were treated with total glucosides of Cistanche deserticola simultaneously. Learning and memory function was tested by Morris water maze in three groups at the 2nd, 4th, 6th and 8th week respectively. The expressions of superoxide dismutase (SOD) and malondialdehyde (MDA) in hippocampus were detected by test kit. Results Compared with the blank control group, the escape latency time of the rats in the IH group and the TGs intervention group was significantly prolonged at the 2nd, 4th, 6th and 8th week respectively (P<0.05). The time to cross the target quadrant in the IH group and the TGs intervention group was gradually shortened at the 2nd, 4th, 6th and 8th week respectively (P<0.05). Compared with the IH group, the escape latency gradually shortened at the 2nd, 4th, 6th and 8th week in the TGs intervention group (P<0.05), while the time to cross the target quadrant was gradually prolonged at the 2nd, 4th, 6th and 8th week (P<0.05). The expressions of MDA in hippocampal tissue in the IH group and the TGs intervention group increased at the 2nd, 4th, 6th and 8th week (P<0.05), which were significantly higher than those in the blank control group; and the expressions of SOD at the 2nd, 4th, 6th, and 8th week were all lower than those in the blank control group(P<0.05). Compared with the IH group, the expression of MDA protein in hippocampal tissue in the TGs intervention group decreased at the 2nd, 4th, 6th and 8th week, while the expression of SOD protein at the 2nd, 4th, 6th and 8th week increased, and the differences were statistically significant (P<0.05). Conclusion The total glycosides of Cistanche deserticola can improve the learning and memory function of intermittent hypoxia rats by inhibiting oxidative stress.