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1.
Article in Chinese | WPRIM | ID: wpr-911296

ABSTRACT

Objective:To evaluate the effect of hydrogen-rich saline on serine threonine protein kinase (Akt) /nuclear factor E2-related factor 2 (Nrf2) signaling pathway during hypoxia/reoxygenation (H/R) injury to human renal tubular epithelial cells.Methods:Human renal tubular epithelial cell line were seeded in 96-well plates at a density of 1.5×10 4 cells/ml (200 μl/well) or in 6-well plates at a density of 2×10 5 cells/ml (2 ml/well) were divided into 5 groups( n=30 each) using a random number table method: control group (group C), hydrogen-rich group (group H), group H/R, H/R plus hydrogen-rich saline group (group H/R+ H) and H/R plus hydrogen-rich saline plus Akt inhibitor uprosertib group (group H/R+ H+ U) .In group C, the cells were incubated for 28 h in an incubator filled with normoxia at 37 ℃ (5%CO 2-21%O 2-74%N 2). In group H, cells were added to the medium containing 0.6 mmol/L hydrogen-rich saline, and then incubated for 28 h in an incubator filled with normoxia at 37 ℃.In group H/R, the cells were incubated in an anaerobic chamber (37 ℃, 5%CO 2-1%O 2-94 %N 2) for 24 h, and then incubated for 4 h in an incubator filled with normoxia at 37 ℃.In group H/R+ H, the cells were incubated in an anaerobic chamber for 24 h, and then incubated for 4 h in an incubator containing 0.6 mmol/L filled with normoxia at 37 ℃.In group H/R+ H+ U, the cells were incubated for 1 h in the culture medium containing uprosertib 10 μmol/L (final concentration) and the other treatments were similar to those previously described in group H/R+ H. After treatment in each group, the cell viability was measured by MTT assay, cell apoptosis was measured using flow cytometry, superoxide dismutase (SOD) activity was measured using xanthine oxidase method), malondialdehyde (MDA) content was detected by thiobarbituric acid method, the expression of Akt, phosphorylated Akt (p-Akt), total Nrf2, nuclear Nrf2 and activated caspase-3 was detected by Western blot, and the expression of Nrf2 mRNA was detected by Real-time PCR. Results:Compared with group C, the cell viability and activity of SOD were significantly decreased, the apoptosis rate and content of MDA were increased, and the expression of p-Akt, nuclear Nrf2, total Nrf2, activated caspase-3 protein and Nrf2 mRNA was up-regulated in group H/R and group H/R+ H ( P<0.05). Compared with group H/R, the cell viability and activity of SOD were significantly increased, the apoptosis rate and content of MDA were decreased, the expression of p-Akt, nuclear Nrf2, total Nrf2 and Nrf2 mRNA was up-regulated and expression of activated caspase-3 protein was down-regulated in group H/R+ H ( P<0.05). Compared with group H/R+ H, the cell viability and activity of SOD was significantly decreased, the apoptosis rate and content of MDA were increased, the expression of p-Akt, nuclear Nrf2, total Nrf2 protein and Nrf2m RNA was down-regulated, and the expression of activated caspase-3 protein was up-regulated in group H/R+ H+ U ( P<0.05). Conclusion:The mechanism by which hydrogen-rich saline attenuates H/R injury to human renal tubular epithelial cells is related to improving activation of Akt/Nrf2 signaling pathway, decreasing oxidative stress response and inhibiting cell apoptosis.

2.
Article in Chinese | WPRIM | ID: wpr-911275

ABSTRACT

Objective:To evaluate the effect of electroacupuncture (EA) pretreatment on oxidative stress response of hippocampus in rats with sepsis-associated encephalopathy (SAE) and the relationship with nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (Nrf2/HO-1) signaling pathway.Methods:A total of 64 healthy adult male Sprague-Dawley rats, weighing 250-300 g, were divided into 4 groups ( n=16 each) using a random number table method: sham operation group (Sham group), SAE group, SAE+ EA group and SAE plus sham EA group (SAE+ SEA group). In SAE+ EA group, Baihui, Quchi and Zusanli acupoints were stimulated for 30 min once a day for 5 consecutive days.Sepsis was induced by cecal ligation and puncture immediately after the end of the last EA.At 1 and 7 days after establishment of the model, the hippocampal malondialdehyde (MDA) and superoxide dismutase (SOD) levels were detected by enzyme-linked immunosorbent assay, the expression of hippocampal Nrf2 mRNA was detected using real-time reverse transcription polymerase chain reaction, and the expression of Nrf2 and HO-1 was determined by Western blot.At 3-7 days after establishment of the model, cognitive function was assessed using the Morris water maze test, and the escape latency and the target quadrant exploration time were recorded. Results:Compared with Sham group, the content of MDA was significantly increased and the activity of SOD was decreased at 1 and 7 days after establishment of the model, the expression of Nrf2 protein and mRNA and HO-1 was down-regulated at day 7 after establishment of the model, the escape latency was prolonged, and the target quadrant exploration time was shortened in SAE group ( P<0.05). Compared with SAE group, the content of MDA was significantly decreased and the activity of SOD was increased at 1 and 7 days after establishment of the model, the expression of Nrf2 protein and mRNA and HO-1 was up-regulated at day 7 after establishment of the model, the escape latency was shortened, and the target quadrant exploration time was prolonged in group SAE+ EA ( P<0.05), and no significant change was found in the parameters mentioned above in group SAE+ SEA ( P>0.05). Conclusion:EA pretreatment can reduce oxidative stress response of hippocampus in rats with SAE, and the mechanism may be related to activating Nrf2/HO-1 signaling pathway.

3.
Article in Chinese | WPRIM | ID: wpr-911259

ABSTRACT

Objective:To evaluate the role of nuclear factor NF-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in atorvastatin-induced reduction of intestinal ischemia-reperfusion (I/R) injury in mice.Methods:Twenty-four healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divide into 4 groups ( n=6 each) using a random number table method: sham operation group (S group), intestinal I/R group (I/R group), atorvastatin group (ATV group) and atorvastatin+ Nrf2 inhibitor ML385 group (AM group). Intestinal I/R was produced by occlusion of superior mesenteric artery for 45 min followed by reperfusion.In ATV and AM groups, atorvastatin 10 mg/kg was given by gavage for 3 consecutive days daily at 3 day before establishment of the model, while the equal volume of normal saline was given by gavage in S and I/R groups.Nrf2 inhibitor ML385 30 mg/kg was intraperitoneally injected at 1 h before establishment of the model in group AM.The mice were sacrificed at 2 h of reperfusion, and intestine tissues were obtained for examination of the pathological changes of intestinal tissues (with a light microscope) which were scored according to Chiu, for determination of wet/dry weight ratio (W/D ratio), for detection of the activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) (by xanthine oxidase method and thiobarbituric acid condensation method) and for determination of the expression of Nrf2 and HO-1 (by Western blot). Results:Compared with S group, the Chiu score, W/D ratio and MDA content were significantly increased, the activity of SOD was decreased, and the expression of Nrf2 and HO-1 was up-regulated in the other 3 groups ( P<0.05). Compared with the group I/R, the Chiu score, W/D ratio and MDA content were significantly decreased, the SOD activity was increased, and the expression of Nrf2 and HO-1 was up-regulated ( P<0.05), and the pathological changes were significantly attenuated in group ATV, and no significant change was found in the parameters mentioned above in AM group ( P>0.05). Compared with the group ATV, the Chiu score, W/D ratio and MDA content were significantly increased, the SOD activity was decreased, the expression of Nrf2 and HO-1 was decreased ( P<0.05), and the pathological changes were significantly aggravated in group AM. Conclusion:The mechanism by which atorvastatin reduces intestinal I/R injury is related to activating Nrf2/HO-1 signaling pathway in mice.

4.
Article in Chinese | WPRIM | ID: wpr-911243

ABSTRACT

Objective:To evaluate the role of silencing information regulator 1 (SIRT1)/nuclear factors E2-related factor2 (Nrf2) signaling pathway in berberine-induced reduction of renal ischemia-reperfusion (I/R) injury in mice.Methods:Thirty SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (S group), renal I/R group (RIR group), berberine+ I/R group (B group), berberine+ I/R+ SIRT1 inhibitor EX527 group (BE group) and berberine+ I/R+ Nrf2 inhibitor ATRA group (BA group). After the right kidney was removed, the left renal artery was clamped for 45 min followed by reperfusion to establish the model of renal I/R injury.In B, BE, and BA groups, berberine 100 mg·kg -1·d -1 was given for intragastric administration at 14 days before surgery.In group BE and group BA, EX527 5 mg·kg -1·d -1 and ATRA 10 mg·kg -1·d -1 were injected intraperitoneally at 3 days before surgery, respectively.The equal volume of normal saline was given for 14 consecutive days in group S and group RIR.Blood samples were collected from orbital vein at 24 h of reperfusion for measurement of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations, for determination of the interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) contents (by enzyme-linked immunosorbent assay) and expression of SIRT1, Nrf2, apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, nucleotide-binding oligomerization domain-like receptor containing pyrin domain (NLRP3) (by Western blot) and for examination of the pathological changes of renal tubules (with a light microscope). The damage to the renal tubules was scored. Results:Compared with group S, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly increased in RIR, B, BE and BA groups, the expression of SIRT1, Nrf2, ASC, caspase-1 and NLRP3 was up-regulated in RIR, BE and BA groups, and the expression of SIRT1, Nrf2, caspase-1 and NLRP3 was up-regulated in group B ( P<0.05). Compared with group RIR, the concentrations of serum Cr and BUN, the contents of renal IL-1β and TNF-α and renal tubular injury score were significantly decreased in B, BE and BA groups, the expression of SIRT1 and Nrf2 in group B, Nrf2 and ASC in BE group and SIRT1, ASC and caspase-1 in BA group was up-regulated, and the expression of ASC, caspase-1 and NLRP3 in group B, SIRT1 and NLRP3 in BE group and Nrf2 in BA group was down-regulated ( P<0.05). Compared with group B, the serum concentrations of Cr and BUN, the contents of IL-1β and TNF-α and renal tubular injury score were significantly increased in BE and BA groups, the expression of ASC, caspase-1 and NLRP3 in BE and BA groups was up-regulated, and the expression of SIRT1 in BE and Nrf2 in BA groups was down-regulated ( P<0.05). Conclusion:SIRT1/Nrf2 signaling pathway is involved in the process of berberine-induced reduction of renal I/R, which is related to inhibiting pyroptosis in mice.

5.
Article in Chinese | WPRIM | ID: wpr-911224

ABSTRACT

Objective:To evaluate the effect of chicoric acid on oxidative stress during myocardial injury in sepsis rats and the relationship with nuclear factor E2-related factor 2 (Nrf2) signaling pathway.Methods:Forty healthy male Sprague-Dawley rats, aged 8-12 weeks, weighing 220-250 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), lipopolysaccharide (LPS) group (group LPS), LPS+ chicoric acid group (group LPS+ CA), LPS+ Nrf2 inhibitor ML385 group (group LPS+ ML) and LPS+ chicoric acid+ ML385 group (group LPS+ CA+ ML). LPS 15 mg/kg was intraperitoneally injected to induce sepsis.Immediately after intraperitoneal injection of LPS, chicoric acid 10 mg/kg or ML385 15 mg/kg (in dimethyl sulfoxide) was intraperitoneally injected in group LPS+ CA and group LPS+ ML, respectively, and ML385 15 mg/kg and chicoric acid 10 mg/kg were intraperitoneally injected in LPS+ CA+ ML group.The equal volume of dimethyl sulfoxide was given instead in group C. At 48 h after establishment of the model, blood samples were collected from the aorta for measurement of concentration of serum interleukin-6 (IL-6) and the activities of lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB) (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and myocardial tissues were obtained for microscopic examination of pathological changes (by HE staining), for determination of activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) and contents of reactive oxygen species(ROS) and iron (by colorimetry), for calculation of the ratio of oxidized nicotinamide adenine 2 nucleotides to reduced nicotinamide adenine 2 nucleotides (NAD + /NADH), and for detection of the expression of Nrf2, NADPH quinone oxidoreductase 1 (NQO1), glutathione peroxidase 4 (GPX4) and nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1) (by Western blot). Results:Compared with C group, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the contents of ROS and iron and the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, and NOX1 expression was up-regulated in the other four groups ( P<0.05). Compared with group LPS, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly decreased, the contents of ROS and iron and the ratio of NAD + /NADH were decreased, activities of GSH-Px and SOD were increased, expression of Nrf2, NQO1 and GPX4 was up-regulated, NOX1 expression was down-regulated ( P<0.05), and the pathological changes of cardiomyocytes were significantly reduced in group LPS+ CA, and the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, NOX1 expression was up-regulated ( P<0.05), and the pathological changes of cardiomyocytes were accentuated in group LPS+ ML.Compared with group LPS+ CA, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the contents of ROS and iron and the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, NOX1 expression was up-regulated ( P<0.05), and the pathological changes of cardiomyocytes were accentuated in group LPS+ CA+ ML. Conclusion:The mechanism by which chicoric acid reduces myocardial injury in sepsis rats may be related to activating Nrf2 signaling pathway and inhibiting oxidative stress.

6.
Article in Chinese | WPRIM | ID: wpr-911199

ABSTRACT

Objective:To evaluate the role of nuclear factor E2-related factor 2 (NRF2) in myocardial ischemia-reperfusion (I/R) injury and the relationship with ferroptosis in diabetic rats.Methods:Forty-eight SPF healthy adult male Sprague-Dawley rats, aged 6-8 weeks, weighing 200-220 g, were divided into 5 groups by a random number table method: sham operation group (group S, n=6), myocardial I/R group (group NIR, n=12), diabetes mellitus+ sham operation group (group DS, n=6), diabetes mellitus+ myocardial I/R group (group DIR, n=12) and diabetes mellitus+ myocardial I/R+ NRF2 agonist sulforaphane group (group DIR+ SFN, n=12). Type 1 diabetes mellitus was induced by intraperitoneal injection of 1% streptozotocin-citrate buffer 60 mg/kg.Sulforaphane 500 μg·kg -1·d -1 was injected intraperitoneally before ischemia for 3 consecutive days in group DIR+ SFN.At the 8th week after establishing the model, myocardial I/R was produced by occlusion of the anterior descending branch of the left coronary artery for 30 min followed by reperfusion.At 2 h of reperfusion, the left ventricular systolic pressure (LVSP), HR, and the maximum rate of increase and decrease of left ventricular systolic pressure (±dp/dt max) were recorded.Blood samples were taken from the carotid artery and the animals were then sacrificed for determination of concentration of cardiac troponin I (cTnI) in serum (using enzyme-linked immunosorbent assay), myocardial Fe 2+ and malondialdehyde (MDA) contents, superoxide dismutase (SOD) activity (by colorimetry) and myocardial infarct size (using TTC) and for determination of expression of NRF2, ferroportin1 (FPN1) and acyl-CoA synthetase long-chain family member 4 (ACSL4) (by Western blot), and the pathological changes of lung tissues were observed by hematoxylin-eosin staining. Results:Compared with group S, LVSP, HR, and ±dp/dt max were significantly decreased, serum cTnI concentration and myocardial Fe 2+ and MDA contents were increased, SOD activity was decreased, expression of ACSL4 was up-regulated and expression of NRF2 and FPN1 was down-regulated in group NIR ( P<0.05). Compared with group DS, LVSP, HR, and ±dp/dt max were significantly decreased, serum cTnI concentration and myocardial Fe 2+ and MDA contents were increased, SOD activity was decreased, expression of ACSL4 was up-regulated and expression of NRF2 and FPN1 was down-regulated in group DIR ( P<0.05). Compared with group NIR, LVSP, HR, and ±dp/dt max were significantly decreased, serum cTnI concentration and myocardial Fe 2+ and MDA contents were increased, SOD activity was decreased, myocardial infarct size was increased, expression of ACSL4 was up-regulated and expression of NRF2 and FPN1 was down-regulated in group DIR ( P<0.05). Compared with group DIR, LVSP, HR, and ±dp/dt max were significantly increased, serum cTnI concentration and myocardial Fe 2+ and MDA contents were decreased, SOD activity was increased, myocardial infarct size was decreased, expression of ACSL4 was down-regulated and expression of NRF2 and FPN1 was up-regulated in group DIR+ SFN ( P<0.05). Conclusion:NRF2 is involved in the process of myocardial I/R injury, which is related to promoting ferroptosis in diabetic rats

7.
Acta cir. bras ; 36(6): e360607, 2021. tab, graf
Article in English | LILACS-Express | MEDLINE, LILACSEXPRESS, LILACS, VETINDEX | ID: biblio-1284911

ABSTRACT

ABSTRACT Purpose To investigate the role of Nrf2/HO-1 in renal histopathological ailments time-dependently in asphyxial cardiac arrest (CA) rat model. Methods Eighty-eight Sprague Dawley male rats were divided into five groups of eight rats each. Asphyxial CA was induced in all the experimental rats except for the sham group. The rats were sacrificed at 6 hours, 12 hours, one day and two days post-CA. Serum blood urea nitrogen (BUN), creatinine (Crtn) and malondialdehyde from the renal tissues were evaluated. Hematoxylin and eosin and periodic acid-Schiff staining were done to evaluate the renal histopathological changes in the renal cortex. Furthermore, Nrf2/HO-1 immunohistochemistry (ihc) and western blot analysis were performed after CA. Results The survival rate of rats decreased in a time-dependent manner: 66.6% at 6 hours, 50% at 12 hours, 38.1% in one day, and 25.8% in two days. BUN and serum Crtn markedly increased in CA-operated groups. Histopathological ailments of the renal cortical tissues increased significantly from 6 hours until two days post-CA. Furthermore, Nrf2/HO-1 expression level significantly increased at 6 hours, 12 hours, and one day. Conclusions The survival rate decreased time-dependently, and Nrf/HO-1 expression increased from 6 hours with the peak times at 12 hours, and one day post-CA.

8.
Chinese Journal of Endemiology ; (12): 441-447, 2021.
Article in Chinese | WPRIM | ID: wpr-909029

ABSTRACT

Objective:To explore the effects of Ginkgo biloba on regulating NF-E2-related factor 2 (Nrf2)-Kelch-like ECH-associated protein 1 (Keap1)-antioxidant response element (ARE) signaling pathway in liver injury induced by coal-burning-borne endemic arsenic poisoning in rats.Methods:Group design method was adopted, according to body weight (80-100 g), a total of 30 Wistar rats were divided into 5 groups (6 rats in each group, half males and half females) by random number table method. The normal control group was fed with normal diet ad libitum for 4.5 months; the Ginkgo biloba control group was fed with Ginkgo biloba (25 mg/kg, 6 d/week) for 1.5 months after normal feeding for 3 months; the drinking water arsenic poisoning group and the arsenic contaminated grain group were fed respectively with 100 mg/L arsenic trioxide (As 2O 3) solution and 100 mg/kg arsenic-containing feed for 3 months, and then fed with normal diet for 1.5 months; the Ginkgo biloba treatment group was fed with 100 mg/kg arsenic-containing feed for 3 months, and then was given Ginkgo biloba (25 mg/kg, 6 d/week) for 1.5 months. After sacrificing the animals, the content of malondialdehyde (MDA), the activity of copper zinc superoxide dismutase (SOD1) and the activity of glutathione peroxidase (GPx) in serum were detected by thiobarbituric acid colorimetry, xanthine oxidase method and dimercaptodinitrobenzoic acid reduction method, respectively. The mRNA and protein expressions of indicator genes of Nrf2-Keap1-ARE signaling pathway in liver tissues were detected by quantitative real-time PCR, immunohistochemistry and Western blotting. Correlation between the indexes was analyzed by Pearson. Results:In drinking water arsenic poisoning group, arsenic contaminated grain group and Ginkgo biloba treatment group, the contents of MDA in serum were (3.54±0.51), (3.83±0.87) and (2.93±0.84) μmol/L, respectively, which were higher than that in normal control group [(1.85±0.36) μmol/L, P < 0.05]; and SOD1 activities [(68.21±4.37), (64.53±9.96), (73.09±5.43) U/ml] and GPx activities [(486.41±40.45), (458.24±42.25), (539.79±79.43) U/L] in serum were lower than those in normal control group [(81.47±5.73) U/ml, (747.86±80.33) U/L, P < 0.05]. Compared with the arsenic contaminated grain group, the content of MDA in serum in Ginkgo biloba treatment group was decreased, the activities of SOD1 and GPx in serum were increased ( P < 0.05). Compared with normal control group, the mRNA expressions of SOD1 and GPx1 in the liver tissues in drinking water arsenic poisoning group, arsenic contaminated grain group and Ginkgo biloba treatment group were significantly higher ( P < 0.05). Compared with arsenic contaminated grain group, the mRNA expressions of SOD1 and GPx1 in the liver tissue in Ginkgo biloba treatment group were increased ( P < 0.05). Compared with the normal control group, the protein expression of SOD1 in liver tissue in arsenic contaminated grain group was decreased ( P < 0.05), the protein expressions of GPx1 were decreased in the liver tissues in drinking water arsenic poisoning group, arsenic contaminated grain group and Ginkgo biloba treatment group ( P < 0.05). Compared with the arsenic contaminated grain group, the protein expressions of SOD1 and GPx1 were increased in the liver tissue in Ginkgo biloba treatment group ( P < 0.05). Compared with the normal control group and arsenic contaminated grain group, the protein expression of Keap1 was decreased in the liver tissue in Ginkgo biloba treatment group ( P < 0.05). Compared with the normal control group, the protein expressions of Nrf2 and phosphorylation of Nrf2 (pNrf2) were increased in the cytoplasm in drinking water arsenic poisoning group, arsenic contaminated grain group and Ginkgo biloba treatment group ( P < 0.05). Compared with the arsenic contaminated grain group, the protein expression of pNrf2 was decreased in the cytoplasm in Ginkgo biloba treatment group ( P < 0.05). The protein expressions of Nrf2 and pNrf2 in the nucleus in drinking water arsenic poisoning group, arsenic contaminated grain group and Ginkgo biloba treatment group were also higher than those in normal control group ( P < 0.05). Compared with the arsenic contaminated grain group, the protein expressions of Nrf2 and pNrf2 were increased in the nucleus in Ginkgo biloba treatment group ( P < 0.05). The results of correlation analysis revealed that the protein expressions of Nrf2 and pNrf2 in the nucleus were negatively correlated with Keap1 protein expression ( r=-0.523,-0.401, P < 0.05), and positively correlated with the mRNA expressions of SOD1 and GPx1 ( r=0.658, 0.530, 0.555, 0.603, P < 0.05). In addition, the protein expressions of SOD1 and GPx1 were positively correlated with their enzyme activities ( r=0.472, 0.629, P < 0.05). Conclusions:Arsenic could induce oxidative stress and liver injury. Ginkgo biloba could reduce the protein expression of Keap1, and promote nuclear translocation of Nrf2, which might induce the up-regulation of mRNA expressions of SOD1 and GPx1, and partially reverse the posttranscriptional regulation of arsenic on SOD1 and GPx1, and then increase their protein expressions and enzyme activities, thereby improve arsenic induced oxidative stress and liver injury.

9.
Acta Pharmaceutica Sinica B ; (6): 1400-1411, 2021.
Article in English | WPRIM | ID: wpr-888811

ABSTRACT

A major mitochondrial enzyme for protecting cells from acetaldehyde toxicity is aldehyde dehydrogenase 2 (ALDH2). The correlation between ALDH2 dysfunction and tumorigenesis/growth/metastasis has been widely reported. Either low or high ALDH2 expression contributes to tumor progression and varies among different tumor types. Furthermore, the ALDH2∗2 polymorphism (rs671) is the most common single nucleotide polymorphism (SNP) in Asia. Epidemiological studies associate ALDH2∗2 with tumorigenesis and progression. This study summarizes the essential functions and potential ALDH2 mechanisms in the occurrence, progression, and treatment of tumors in various types of cancer. Our study indicates that ALDH2 is a potential therapeutic target for cancer therapy.

10.
Article in Chinese | WPRIM | ID: wpr-871703

ABSTRACT

Objective To investigate the inhibitory effect oflentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygeninduced retinopathy (OIR).Methods One hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group,simple OIR model group,OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group),with 16,32,32 and 32 mice,respectively.When the mice were 7 days old,the mice in the normal control group were fed in a routine environment,and the mice in the OIR model group,Vec group and PSF group were established OIR model.The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1 × 10~ TU/ml) at the age of 12 days.No injection was performed in the normal control group and simple OIR group.RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina.Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase1 (HO-1).Western blot analysis was applied to detect the protein expression ofNrf2,HO-1 and PSF.Results Of the normal control group,simple OIR model group,Vec group and PSF group,the number of pre-retinal neovascular cell nuclei were 0.00,14.36 ± 5.50,15.67 ± 4.96,8.13 ± 2.09,the non-perfusion area were 0.00%,(35.71 ± 2.81)%,(36.57 ± 4.53)%,(15.33 ± 4.75)%,respectively.The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87,165.70;P<0.05).Compared with the normal control group,there were more pre-retinal neovascular cell nucleis and larger nonperfusion area in the simple OIR model group and Vec group (P<0.05).Compared with the simple OIR model group and Vec group,there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P<0.05).Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2,HO-1 (F=53.66,83.54) and protein expression ofNrf2,HO-1 and PSF (F=58.38,52.69,24.79) among 4 groups were significant (P<0.05).The rnRNA expression ofNrf2,HO-1 and protein expression of Nrf2,HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P<0.05).The mRNA expression ofNrf2,HO-1 and protein expression ofNrf2,HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P<0.05).model group and Vec group (P<0.05).Conclusion Intravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.

11.
Chinese Journal of Dermatology ; (12): 128-132, 2020.
Article in Chinese | WPRIM | ID: wpr-870235

ABSTRACT

Objective To evaluate the protective effect of nuclear factor E2-related factor 2(Nrf2) protein against ultraviolet B (UVB)-induced photodamage to HaCaT cells,and to explore its mechanisms.Methods Cultured HaCaT cells were divided into 4 groups:control group receiving no treatment,UVB group irradiated with 30 mJ/cm2 UVB for 30 s,Nrf2 group transfected with a lentiviral vector overexpressing the Nrf2 gene,and Nrf2 + UVB group transfected with a lentiviral vector overexpressing the Nrf2 gene followed by radiation with 30 mJ/cm2 UVB for 30 s.After the treatment,HaCaT cells in the above 4 groups were cultured for another 24 hours.Then,changes in the morphology of HaCaT cells were observed after UVB radiation,Western blot analysis was performed to determine Nrf2 protein expression,cell counting kit-8 (CCKS) assay to detect survival rates of HaCaT cells,flow cytometry to detect levels of reactive oxygen species (ROS),and a biochemical method to detect superoxide dismutase (SOD) levels in cells,and enzyme-linked immunosorbent assay to detect levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α in the culture supematant of HaCaT cells.One-way analysis of variance was used for comparing means in several groups,and least significant difference (LSD)-t test for multiple comparisons.Results Polygonal and clustered HaCaT cells were observed in the control group.After UVB radiation,HaCaT cells became shrunken and round,the number of floating cells increased,and the number of adherent cells markedly decreased.There was a significant difference in Nrf2 protein expression among the control group,UVB group,Nrf2 group and Nrf2 + UVB group (1.84 ± 0.047,0.63 ± 0.082,2.19 ± 0.168 and 1.43 ± 0.069 respectively;F =64.81,P < 0.05),and the Nrf2 protein expression was significantly higher in the Nrf2 group than in the control group (t =14.82,P < 0.05);the survival rates of HaCaT cells also significantly differed among the above 4 groups (98.00% ± 2.39%,24.40% ± 2.98%,71.63% ± 3.39%and 43.38% ± 3.39% respectively;F =236.66,P < 0.05),and the UVB group showed significantly decreased cell viability compared with the control group (t =33.34,P < 0.05)and Nrf2 + UVB group (t=10.07,P < 0.05);a significant difference in the ROS level in HaCaT cells was observed among the above 4 groups (1.27 ± 0.10,5.65 ± 0.19,2.10 ± 0.73 and 3.67 ± 0.19 respectively;F =481.39,P < 0.05),and the UVB group showed a significantly increased ROS level compared with the control group (t =33.68,P <0.05) and Nrf2 + UVB group (t =12.47,P < 0.05).The SOD level in HaCaT cells significantly differed among the above 4 groups (F =170.76,P < 0.05),and was significantly lower in the UVB group than in the control group (t =11.25,P < 0.05) and Nrf2 + UVB group (t =17.52,P < 0.05).The IL-6 level also significantly differed among the above 4 groups (F =532.34,P < 0.05),and was significantly higher in the UVB group than in the control group (t =28.48,P < 0.05) and Nrf2 + UVB group (t =27.82,P < 0.05).There was no significant difference in the TNF-α level among the above 4 groups (F =2.02,P =0.19).Conclusion Nrf2 can protect HaCaT cells from UVB-induced oxidative damage,by reducing intracellular ROS levels and increasing the activity of the endogenous antioxidant enzyme SOD.

12.
Journal of Clinical Hepatology ; (12): 924-927, 2020.
Article in Chinese | WPRIM | ID: wpr-819199

ABSTRACT

Nonalcoholic fatty liver disease (NAFLD) is the most prevalent chronic liver disease around the world, and its incidence rate is increasing year by year. Oxidative stress is the second hit in the classic “two-hit” pathogenesis of NAFLD, which is currently recognized as one of the pathogeneses of NAFLD. Nuclear factor erythroid 2-related factor 2 (Nrf2) is a group of positive regulators that protect hepatocytes against oxidative stress. It is a key factor for cellular anti-oxidative stress and a key transcription factor that antagonizes liver oxidative stress. It plays an important role in the development and progression of NAFLD and may be a potential treatment target for improving NAFLD. This article reviews the role of oxidative stress and the Nrf2 pathway in the pathogenesis of NAFLD.

13.
Rev Assoc Med Bras (1992) ; 66(7): 986-991, 2020. graf
Article in English | SES-SP, LILACS, SES-SP | ID: biblio-1136303

ABSTRACT

SUMMARY OBJECTIVE Nuclear factor erythroid-2 related factor 2 (Nrf2)/ antioxidant response element (ARE) is a novel defensive pathway involved in the oxidative and chemical stress of cells. The aim of the study was to explore the role of Nrf2 on the apoptosis of human disc nucleus pulpous cells induced by hydrogen peroxide (H2O2). METHODS The degeneration model of human intervertebral disc nucleus pulpous cells was established. The expression of Nrf2 was interfered with using sulforaphane (SFN); for that end, three groups were established: a blank group (H2O2-/SFN-), control group (H2O2+/SFN-), and an experimental group (H2O2+/SFN+). CCK8, Hoechst 33258 living cell staining was used to detect reactive oxygen species (ROS) content. RESULTS The apoptotic rates of the three groups were [(0.40±0.46)%], [(25.98±11.28)%], and [(3.83±2.06)%, respectively. The difference was statistically significant (p<0.05). The relative content of ROS in the three groups was [(100±7)%], [(1538±91)%], and [(818±63)%]; the difference was statistically significant (p<0.05). In Western blotting, Nrf2 content in the experimental group was higher than that in the control group. CONCLUSION Nrf2 exists in the nucleus pulpous cells of human intervertebral discs, which is related to the degeneration of the intervertebral disc. It has negative feedback regulation and can prevent the degeneration of the intervertebral disc by inhibiting the apoptosis of nucleus pulpous cells of human intervertebral discs caused by excessive ROS, which provides a new intervention strategy for the prevention and treatment of the degeneration of intervertebral discs.


RESUMO OBJETIVO O fator 2 relacionado a NF-E2 (Nrf2)/elemento de resposta antioxidante (ARE) é uma nova via defensiva envolvida no estresse oxidativo e químico das células. O objetivo deste estudo foi explorar o papel do Nrf2 na apoptose das células do núcleo pulposo do disco humano induzida pelo peróxido de hidrogênio (H2O2). MÉTODOS O modelo de degeneração das células do núcleo pulposo do disco intervertebral humano foi estabelecido. A expressão do Nrf2 foi interferida utilizando-se sulforafano (SFN). Para isso foram estabelecidos três grupos: um grupo vazio (H2O2-/SFN-), um grupo de controle (H2O2+/SFN-), e um grupo experimental (H2O2+/SFN+). Utilizando CCK8 e Hoechst 33258, o conteúdo de espécies reativas de oxigênio (ERO) foi detectado. RESULTADOS As taxas de apoptose dos três grupos foram [(0,40 ± 0,46)%], [(25,98 ± 11,28%)] e [(3,83 ± 2,06)%], respectivamente. A diferença apresentou significância estatística (p < 0,05). O conteúdo relativo de ERO nos três grupos foi [(100±7)%], [(1538±91%)], e [(818±63%); a diferença foi estatisticamente significativa (p < 0,05). O método de Western blotting indicou um maior conteúdo de Nrf2 no grupo experimental do que no grupo de controle. CONCLUSÃO O Nrf2 existe em células do núcleo pulposo do disco intervertebral humano, que estão relacionadas à degeneração do disco intervertebral. Ele apresenta regulação por feedback negativo e pode evitar a degeneração do disco intervertebral inibindo a apoptose de células do núcleo pulposo do disco causada por excesso de ERO. Essa informação proporciona uma nova estratégia de intervenção para a prevenção e o tratamento da degeneração do disco intervertebral.


Subject(s)
Humans , Apoptosis , Oxidative Stress , NF-E2-Related Factor 2 , Intervertebral Disc Degeneration/metabolism , Hydrogen Peroxide
14.
Article in Chinese | WPRIM | ID: wpr-857539

ABSTRACT

OBJECTIVE To investigate the effects of bilobalide (BB) on the myocardial tissue of rats with myocardial ischemia/reperfusion (Ml/R). METHODS An Ml/R model was prepared by ligating the anterior descending branch of the left coronary artery in rats for 30 min. BB 2, 4 and 8 mg-kg"1 was adminsterted for four weeks. The heart rate of rats was recorded and myocardial infarction area was detected. Detective kits were used to evaluate the concentrations of creatine kinase MB isoenzyme (CK-MB) and lactate dehydrogenase (LDH). HE staining was performed to observe the myocardial injury, and TUNEL staining was performed to evaluate apoptosis. ELISA was performed to determine the concentrations of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), IL-1p, tumor necrosis factor-a (TNF-a) and IL-10. Immunohistochemistry was performed to evaluate the expressions of Ki67 and interleukin-6 (IL-6). Western blotting was performed to evaluate the expressions of Bcl-2, Bax, cleaved-caspase 3, nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2), heme oxygenase 1 (HO-1), NAD(P)H quinone dehydrogenase 1 (NQ01), NF-kB P65, phosphorylated inhibitor of NF-kB (P-IkBoc) and IkBcx kinase (p-IKK). RESULTS Compared with normal control group, the percentage of the myocardial infarction area and the concentration of CK-MB and LDH were increased in Ml/R model group, but heart rate was decreased (P<0.01) and myocardial tissue showed myocardial fiber rupture and inflammatory infiltration. Compared with Ml/R model group, the percentage of the myocardial infarction area and the concentration of CK-MB and LDH were decreased in Ml/R model+BB 2, 4 and 8 mg-kg"1 groups (P<0.05), but heart rate was increased (P<0.01) and myocardial tissue fibers tended to be regular while inflammatory cell infiltration was significantly reduced. Compared with normal control group, the percentage of apoptotic cells and Ki67 expression-positive cells and the expressions of Bax and cleaved-caspase 3 were increased in Ml/R model group (F<0.01), but the expression of Bcl-2 was decreased markedly (P<0.01). Compared with Ml/R model group, the percentages of apoptotic cells and Ki67 expression-positive cells and the expressions of Bax and cleaved-caspase 3 were decreased in Ml/R model+BB 2, 4 and 8 mg∗kg-1 groups (P<0.01), while the expression of Bcl-2 was increased (P<0.01). Ml/R could lower the activity of SOD and GSH-Px, but enhance MDA content. BB could weaken the effect of Ml/R on SOD, GSH-Px and MDA. Compared with normal control group, the concentrations of IL-1β, TNF-α and IL-10 in serum and the percentage of IL-6 expression-positive cells in myocardial tissue of Ml/R model group were increased (P<0.01). Compared with Ml/R model group, the concentrations of IL-1β and TNF-a and the percentage of IL-6 expression-positive cells were decreased in Ml/R model∗ BB 2, 4 and 8 mg-kg"1 group (P<0.01), but the concentration of IL-10 was increased(P<0.05). Compared with mormal control group, the protein levels of Nrf2, HO-1 and NQ01 significantly decreased (P<0.01), while the levels of NF-kB P65, p-IKK and p-kBa significantly increased (P<0.01). BB could effectively reverse protein changes (P<0.05, P<0.01). CONCLUSION BB can protect rats with Ml/R via anti-inflamma-tion and anti-oxidation.

15.
Article in Chinese | WPRIM | ID: wpr-857525

ABSTRACT

NF-E2-related factor 2 (Nrf2) is a member of the CNC basic leucine zipper (b-ZIP) family, which plays a key role in maintaining cell redox homeostasis, regulating energy metabolism, inflammatory response, apoptosis, and autophagy. Mitochondria are the main sites for ATP synthesis, and one of the main sources of reactive oxygen free radicals. Recently, increasing studies have suggested that Nrf2 pathway may play a critical role in regulation of mitochondrial function in various ways and involving multiple mechanisms, thus regulating cellular responses and adverse outcomes to endogenous stimuli. These include regulation of reactive oxygen species levels, inhibition of mitochondrial membrane potential decline, promotion of ATP synthesis, protection of mitochondrial fatty acid oxidation and oxidative phosphorylation, mitochondrial biogenesis, and maintenance of mitochondrial integrity as well as mitochondrial autophagy. In addition, Nrf2 pathway also plays an important role in regulation of toxicant induced mitohormesis. This review summarizes recent research progress in the regulation of mitochondrial function by Nrf2 pathway and its mechanism.

16.
Chinese Journal of Anesthesiology ; (12): 1189-1193, 2019.
Article in Chinese | WPRIM | ID: wpr-824686

ABSTRACT

Objective To evaluate the role of phosphatidylinositol-3-kinase/protein kinase B/nucle-ar factor erythroid 2-related factor 2(PI3K/Akt/Nrf2)signaling pathway in resveratrol preconditioning-in-duced cardioprotection in diabetic rats.Methods Clean-grade healthy male Sprague-Dawley rats,aged 2-3 months,weighing 240-280 g,were used in the study.The diabetes model was established by intraper-itoneal injection of streptozotocin 30 mg/kg once a day for 7 consecutive days.Forty diabetic rats were se-lected and divided into 4 groups(n=10 each)according to the random number table method: sham opera-tion group(S group),myocardial ischemia-reperfusion(I/R)group(I/R group),resveratrol plus myo-cardial I/R group(Res+I/R group),and PI3K inhibitor LY294002 plus resveratrol plus myocardial I/R group(LY+Res+I/R group).The myocardial I/R injury model was established by ligating the left anterior descending coronary artery for 30 min followed by 120-min reperfusion in anesthetized rats.Resveratrol 20 mg/kg was intraperitoneally injected once a day for 7 consecutive days at one week before surgery in Res+I/R and LY+Res+I/R groups.LY294002 1.5 mg/kg was intraperitoneally injected at 30 min before operation in LY+Res+I/R group.After 120 min of reperfusion,blood samples were taken for determination of serum creatine kinase-MB(CK-MB)and lactic dehydrogenase(LDH)concentrations(by enzyme-linked immu-nosorbent assay),and myocardial tissues at the ischemic area were obtained for determination of superoxide dismutase(SOD),glutathione(GSH)and malondialdehyde(MDA)levels(by enzyme-linked immu-nosorbent assay)and expression of Akt,phosphorylated Akt(p-Akt),glycogen synthase kinase 3β(GSK3β),phosphorylated GSK3β(p-GSK3β)and Nrf2(by Western blot).Results Compared with group S,serum CK-MB and LDH concentrations were significantly increased,the SOD and GSH levels were decreased,the MDA level was increased,and the expression of p-Akt and p-GSK3β was down-regu-lated in I/R group(P<0.05).Compared with I/R group,serum CK-MB and LDH concentrations were sig-nificantly decreased,the SOD and GSH levels were increased,the MDA level was decreased,and the ex-pression of p-Akt,p-GSK3β and Nrf2 was up-regulated in Res+I/R group(P<0.05).Compared with Res+I/R group,serum CK-MB and LDH concentrations were significantly increased,the SOD and GSH levels were decreased,the MDA level was increased,and the expression of p-Akt,p-GSK3β and Nrf2 was down-regulated in LY+Res+I/R group(P<0.05).Conclusion The mechanism of resveratrol precondi-tioning-induced cardioprotection is related to activating PI3K/Akt/Nrf2 signaling pathway and inhibiting oxi-dative stress responses in diabetic rats.

17.
International Eye Science ; (12): 268-271, 2019.
Article in Chinese | WPRIM | ID: wpr-713011

ABSTRACT

@#AIM:To investigate the association between NF-E2-related factor 2(Nrf2)gene polymorphism and diabetic retinopathy(DR)in the Han descent population of Shaanxi province. <p>METHODS: Totally 386 patients with type 2 diabetes mellitus(T2DM)who hospitalized in our hospital from January 2016 to January 2017 were recruited to participate in the study, they were divided into diabetic without retinopathy(DWR)group(181 cases), proliferative diabetic retinopathy(PDR)group(62 cases)and non-proliferative diabetic retinopathy(NPDR)group(143 cses). Meanwhile, 120 healthy volunteers without DR were recruited to participate in the study as the control group. Genotype was determined by polymerase chain reaction-restriction fragment length polymorphism combined with DNA direct sequencing technique for the polymorphism of the Nrf2 gene. <p>RESULTS: There was no significant difference of genotype and allele distribution of rs6721961 polymorphism between the DWR group and control group, neither PDR group and NPDR group(<i>P</i>>0.05). PDR group and NPDR group were compared with control group and DWR group respectively, the genotype and allele distribution of Nrf2 polymorphism were significantly different(<i>P</i><0.05). Multivariable logistic regression analysis showed that A allele(<i>OR</i>=1.532, 95%<i>CI</i> 1.169-2.008, <i>P</i>=0.014)distribution of rs6721961 polymorphism were associated with risk of DR. <p>CONCLUSION: Polymorphism of the Nrf2 gene may associate with the risk of DR.

18.
Article in Chinese | WPRIM | ID: wpr-746211

ABSTRACT

Objective To observe the expression ofprobucol on high glucose-induced specificity protein 1 (SP 1),kelchlike ECH associated protein 1 (Keap 1),NF-E2-related factor 2 (Nrf2) and glutamatecysteine ligase catalytic (GCLC) in the cultured human müller cells and preliminary study the antioxidation of the probucol on müller cells.Methods Primary cultured human müller cells were randomly divided into four groups:normoglycaemia group (5.5 mmol/L glucose),normoglycaemia with probucol group (5.5 mmol/L glucose+100 μmol/L probucol),hyperglycemia group (25.0 mmol/L glucose),hyperglycemia with probucol group (25.0 mmol/L glucose + 100 μmol/L probucol).Immunofluorescence staining was used to assess distribution of SP1,Keapl,Nrf2,GCLC in human Müller cells.SP1,Keapl,Nrf2 and GCLC messenger RNA (mRNA) expression was evaluated by quantitative real-time RT-PCR (qRT-PCR).Independent sample t test was used to compare the data between the two groups.Results All müller cells expressed glutamine synthetase (> 95%),which confirmed the cultured cells in vitro were the purification of generations of müller cells.The expressions of SP 1,Keap 1,Nrf2,and GCLC protein were positive in human müller cells.qRT-PCR indicated that SP1 (t=28.30,P<0.000),Keap1 (t=5.369,P=0.006),and Nrf2 (t=10.59,P=0.001) mRNA in the hyperglycemia group increased obviously compared with the normoglycaemia group;GCLC (t=4.633,P=0.010)mRNA in the hyperglycemia group decreased significantly compared with the normoglycaemia group.However,SP1 (t=12.60,P=0.000) and Keapl (t=4.076,P=0.015) in the hyperglycemia with probucol group decreased significantly compared with the hyperglycemia group;Nrf2 (t=12.90,P=0.000) and GCLC (t=l 5.96,P<0.000)mRNA in the hyperglycemia with probucol group increased obviously compared with with the hyperglycemia group.Conclusion Probucol plays an antioxidant role by inhibiting the expression of SP 1,Keap 1 and upregulating the expression of Nrf2,GCLC in müller cells induced by high glucose.

19.
Article in Chinese | WPRIM | ID: wpr-755630

ABSTRACT

Objective To evaluate the effect of hydrogen preconditioning during cold ischemia phase on the activity of nuclear factor erythroid 2-related factor 2 ( Nrf2) in rat pulmonary microvascular en-dothelial cells ( PMVECs) subjected to hypoxia-reoxygenation ( H/R) . Methods PMVECs were isolated from clean-grade male Sprague-Dawley rats, aged 2-3 weeks, using the tissue block adherence method and divided into 4 groups ( n=25 each) using a random number table method: control group ( group C) , H/R group, oxygen group ( O group) and hydrogen group ( H group) . Cells were incubated for 4 h with 4℃ low potassium dextransolution ( LPD) pre-equilibrated with 95% oxygen and 5% carbondioxide to simulate the cold ischemia phase. LPD pre-balanced with 95% oxygen and 5% carbon dioxide was replaced with LPD, and then cells were incubated for 1 h at room temperature to simulate the lung transplantation period. LPD was rapidly replaced with 37℃ M199 complete culture solution, and cells were incubated in the mixture of 40% oxygen-5% carbondioxide-55% nitrogen to simulate the reperfusion period. In O and H groups, the cells were exposed to 40% oxygen-60% nitrogen and 3% hydrogen-40% oxygen-57% nitrogen during the cold ischemia period, respectively, and the gas mixture was replaced every 20 min. The cell culture fluid was collected 4 h later for determination of interleukin ( IL )-6, IL-10 and tumor necrosis factor-alpha ( TNF-α) concentrations ( by enzyme-linked immunosorbent assay) and malondialdehyde ( MDA) concen-trations ( by thiobarbituric acid method) . The cytoplasm and nucleoproteins were extracted for measurement of Nrf2 expression ( by Western blot) and cell apoptosis ( by flow cytometry and TUNEL assay) . The cell apoptosis rate was calculated. Results Compared with C group, the IL-6, TNF-α and MDA levels were significantly increased, the IL-10 level was decreased, the apoptosis rate was increased, and the expres-sion of Nrf2 in nucleus was up-regulated in H/R group ( P>0. 05) . Compared with H/R group, the IL-6, TNF-α and MDA levels were significantly decreased, the IL-10 level was increased, the apoptosis rate was decreased, and the Nrf2 expression in cytoplasm was down-regulated in O and H groups (P<0. 05), the Nrf2 expression was significantly up-regulated in H group ( P<0. 05) , and no significant change was found in the expression of Nrf2 in nucleus in O group ( P>0. 05) . Compared with O group, the IL-6, TNF-αand MDA levels were significantly decreased, the IL-10 level was increased, the apoptosis rate was decreased, the expression of Nrf2 in nucleus was up-regulated, and the expression of Nrf2 protein in cytoplasm was down-regulated in H group ( P<0. 05 ) . Conclusion The mechanism by which hydrogen preconditioning during cold ischemia phase reduces H/R injury to rat PMVECs is related to activating Nrf2 and thus inhibi-ting oxidative stress.

20.
Article in Chinese | WPRIM | ID: wpr-755590

ABSTRACT

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 ( Nrf2 ) signaling pathway in endoplasmic reticulum stress response during lipopolysaccharide ( LPS)-induced acute lung injury ( ALI) in mice. Methods Forty clean-grade healthy male C57BL∕6 mice, aged 6-8 weeks, weighing 22-26 g, were divided into 4 groups ( n=10 each) using a random number table method: control group ( group C) , group ALI, salubrinal group ( group S) and salubrinal plus brusatol group ( group S+B) . Animals were intratracheally instilled with 5 mg∕kg of LPS diluted in normal saline to establish the model of ALI. Animals were intratracheally instilled with 100 μl of normal saline in group C. Mice in group S were intraperitoneally injected with endoplasmic reticulum stress response inhibitor 1 mg∕kg salubrinal at 1 and 24 h after LPS instillation. Mice of group S+B were intraperitoneally injected with brusatol 2 mg∕kg once every other day for 10 days prior to LPS instillation, and the other treatments were similar to those previously de-scribed in group S. Mice were sacrificed at 48 h after LPS administration, and lungs were removed for mi-croscopic examination of the pathological changes of lung tissues which were scored and for determination of contents of IL-17A, tumor necrosis factor-alpha ( TNF-α) and interleukin-6 ( IL-6) in lung tissues ( by en-zyme-linked immunosorbent assay) and expression of Nrf2, CCAAT∕enhancer-binding protein homologous protein (CHOP) and caspase-12 in lung tissues (by Western blot). Lung water content was calculated. Results Compared with group C, the lung water content and contents of IL-17A, TNF-α and IL-6 were significantly increased, the expression of CHOP and caspase-12 in cytoplasma was up-regulated, and the ex-pression of Nrf2 in nuclei was down-regulated in ALI and S+B groups, and the lung water content and con-tents of IL-17A, TNF-α and IL-6 were significantly increased, the expression of Nrf2 in nuclei and CHOP in cytoplasma was up-regulated, and the expression of caspase-12 was down-regulated in group S ( P<0. 05) . Compared with group ALI, the lung water content and contents of IL-17A, TNF-α and IL-6 were significantly decreased, the expression of CHOP and caspase-12 in cytoplasma was down-regulated, the ex-pression of Nrf2 in nuclei was up-regulated ( P<0. 05) , and the pathological changes were significantly at-tenuated in group S. Compared with group S, the lung water content and contents of IL-17A, TNF-α and IL-6 were significantly increased, the expression of CHOP and caspase-12 in cytoplasma was up-regulated, the expression of Nrf2 in nuclei was down-regulated ( P<0. 05) , and the pathological changes of lung tis-sues were accentuated in group S+B. Conclusion Nrf2 signaling pathway is involved in the process of en-doplasmic reticulum stress response during LPS-induced ALI in mice.

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