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Objective:To evaluate the role of the nuclear factor E2-related factor 2 (Nrf2) signaling pathway in hydrogen sulfide (H 2S)-induced inhibition of inflammatory responses in the brain tissues of mice with sepsis-associated encephalopathy (SAE). Methods:Fifty-four wild-type C57BL/6J mice and thirty-six Nrf2 -/-C57BL/6J mice, weighing 20-25 g, were divided into 5 groups ( n=18 each) using a random number table method: wild-type sham operation group (wild-type Sham group), wild-type SAE group, wild-type SAE+ NaHS group, Nrf2 -/-SAE group, and Nrf2 -/-SAE+ NaHS group.After the model of SAE was established by cecal ligation and puncture in anesthetized mice.NaHS 50 μmol/kg was intraperitoneally injected at 3 h after the model was successfully established.The mice were sacrificed at 24 h after surgery, and brain tissues were obtained for examination of the phathological changes and for determination of the number of viable neurons, the expression of NLRP3 (by Western blot), contents of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 (by enzyme-linked immunosorbent assay), and percentage of Iba-1 + CD86 + , Iba-1 + CD206 + and Iba-1 + cells (by flow cytometry). Results:Compared with wild-type Sham group, NLRP3 expression was significantly up-regulated, contents of TNF-α, IL-1β and IL-6 and percentage of Iba-1 + CD86 + and Iba-1 + cells were increased, and the percentage of Iba-1 + CD206 + cells and neuron survival rate were decreased in wild-type SAE group ( P<0.05). Compared with wild-type SAE group, NLRP3 expression was significantly down-regulated, contents of TNF-α, IL-1β and IL-6, percentage of Iba-1 + and Iba-1 + CD86 + and neuron survival rate were decreased, and the percentage of Iba-1 + CD206 + cells was increased in wild-type SAE+ NaHS group ( P<0.05). There was no significant difference in each parameter between Nrf2 -/-SAE group and Nrf2 -/-SAE+ NaHS group ( P>0.05). Compared with wild-type SAE+ NaHS group, NLRP3 expression was significantly up-regulated, the percentage of Iba-1 + and Iba-1 + CD86 + and contents of TNF-α, IL-1β and IL-6 were increased, and the percentage of Iba-1 + CD86 + cells and neuron survival rate were decreased in Nrf2 -/-SAE+ NaHS group ( P<0.05). Conclusion:Nrf2 signaling pathway is involved in H 2S-induced inhibition of inflammatory responses in the brain tissues of mice with SAE.
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Objective:To evaluate the effect of hydrogen-rich saline on the activity of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammomes during renal ischemia-reperfusion (I/R) in mice.Methods:Thirty male C57BL/6J mice, weighing 20-25 g, aged 6-8 weeks, were divided into 5 groups ( n=6 each) using a random number table method: sham operation group (S group), I/R group, I/R plus NLRP3 inhibitor MCC950 group (I/R+ M group), I/R plus hydrogen-rich saline group (I/R+ H group), and I/R plus MCC950 plus hydrogen-rich saline group (I/R+ M+ H group). The model of renal I/R injury was established by clipping the bilateral renal pedicles for 30 min followed by reperfusion in anesthetized animals.MCC950 20 mg/kg was intraperitoneally injected for 14 consecutive days before surgery in I/R+ M and I/R+ M+ H groups.Hydrogen-rich saline 5 ml/kg was intraperitoneally injected at 1 h after surgery in I/R+ H and I/R+ M+ H groups.The equal volume of normal sline was given instead in the other groups.Blood samples from hearts were collected at 24 h of reperfusion for determination of concentrations of blood urea nitrogen (BUN), creatinine (Cr) and kidney injury molecule-1 (Kim-1), tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β) and IL-6 in serum (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and kidney tissues were obtained for microscopic examination of pathological changes of renal tissues (with light microscopes) and for determination of cell apoptosis and expression of NLRP3, apoptosis-associated speck-like protein containing C-terminal caspase recruitment domain (ASC) and caspase-1 protein and mRNA (by Western blot or real-time polymerase chain reaction). Results:Compared with group S, the serum concentrations of Cr, BUN, Kim-1, TNF-α, IL-1β and IL-6 were significantly increased, the kidney injury score and the number of apoptotic cells were increased, and the expression of NLRP3, ASC, caspase-1 protein and mRNA was up-regulated in I/R, I/R+ M, I/R+ H and I/R+ M+ H groups ( P<0.05). Compared with group I/R, the serum concentrations of Cr, BUN, Kim-1, TNF-α, IL-1β and IL-6 were significantly decreased, the kidney injury score and the number of apoptotic cells were decreased, and the expression of NLRP3, ASC, caspase-1 protein and mRNA was down-regulated in I/R+ M, I/R+ H and I/R+ M+ H groups ( P<0.05). Compared with group I/R+ H, the serum concentrations of Cr, BUN, Kim-1 and TNF-α, IL-1β and IL-6 were significantly decreased, the kidney injury score and the number of apoptotic cells were decreased, and the expression of NLRP3, ASC, caspase-1 protein and mRNA was down-regulated in group I/R+ M+ H ( P<0.05). Conclusion:The mechanism by which hydrogen-rich saline reduces renal I/R injury is related to inhibiting the activation of NLRP3 inflammomes in mice.
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Objective:To evaluate the role of hypoxia-inducible factor-1α (HIF-1α) in hydrogen-induced inhibition of lipopolysaccharide (LPS)-induced inflammatory responses in mouse macrophages.Methods:The mouse RAW264.7 macrophages cultured in vitro were divided into 4 groups ( n=24 each) according to the random number table method: control group (C group), LPS group (L group), hydrogen-rich solution plus LPS group (H+ L group), and hydrogen-rich solution plus LPS plus HIF-1α inhibitor 2-methoxyestradiol (2ME2) group (H+ L+ M group). LPS 1 μg/ml was added, and the cells were incubated for 6 h in group L. In group L+ H, LPS was added first, the medium was changed to 0.6 mmol/L hydrogen-rich solution, and cells were incubated for 6 h. In group H+ L+ M, 2ME2 10 μmol/L was given first, cells were then incubated for 30 min, LPS and hydrogen-rich solution were added, and cells were incubated for 6 h. Western blot was used to determine the expression of HIF-1α, Beclin-1, Bcl-2/E1B-19 kDa interacting protein 3 (BNIP3) and LC3.Enzyme-linked immunosorbent assay was used to detect the concentrations of tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β in the supernatant.The number of autophagosomes was observed using a transmission electron microscope. Results:Compared with group C, the concentrations of TNF-α, IL-6 and IL-1β in the supernatant were significantly increased, the expression of HIF-1α, Beclinl and BNIP3 in macrophages was up-regulated, the ratio of LC3Ⅱ/LC3Ⅰ was increased, and the number of autophagosomes was increased in group L ( P<0.05). Compared with group L, the concentrations of TNF-α, IL-6 and IL-1β were significantly decreased, the expression of HIF-1α, Beclin-1 and BNIP3 in macrophages was up-regulated, LC3Ⅱ/LC3Ⅰ ratio was increased, and the number of autophagosomes was increased in group H+ L ( P<0.05). Compared with group H+ L, the concentrations of TNF-α, IL-6 and IL-1β in the supernatant were significantly decreased, the expression of HIF-1α, Beclin-1, and BNIP3 in macrophages was down-regulated, and the ratio of LC3Ⅱ/LC3Ⅰ was decreased, and the number of autophagosomes was decreased in group H+ L+ M ( P<0.05). Conclusion:HIF-1α-mediated activation of autophagy is involved in the process of hydrogen-induced inhibition of LPS-induced inflammatory responses in mouse macrophages.
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Objective:To evaluate the relationship between phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway and heme oxygenase-1 (HO-1) expression during lung injury in septic mice.Methods:Forty-eight clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group SH), sepsis group (group SEP), PI3K inhibitor LY294002 group (group LY) and LY294002+ HO-1 agonist hemin group (group LH). Sepsis was induced by cecal ligation and puncture in anesthetized animals.LY294002 30 mg/kg was intraperitoneally injected at 2 h before establishing the model in LY group and LH group.Hemin 50 μmol/kg was intraperitoneally injected at 1 h before establishing the model in LH group.Mice were sacrificed at 24 h after surgery, and lungs were removed for determination of wet/dry weight ratio (W/D ratio), tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) contents (by enzyme-linked immunosorbent assay), and expression of phosphorylated Akt(p-Akt), Akt and HO-1 (by Western blot) and for examination of the pathological changes of lung tissues which were scored. Results:Compared with SH group, the W/D ratio, lung injury score, and TNF-α and IL-10 contents were significantly increased in SEP, LY and LH groups, and the expression of p-Akt and HO-1 was significantly up-regulated in SEP group ( P<0.05). Compared with SEP group, the W/D ratio, lung injury score and TNF-α content were significantly increased, IL-10 content was decreased, and the expression of p-Akt and HO-1 was down-regulated in LY group ( P<0.05). Compared with LY group, the lung injury score and TNF-α content were significantly decreased, IL-10 content was increased, and the expression of HO-1 was up-regulated in LH group ( P<0.05). Conclusion:PI3K/Akt signaling pathway is involved in the endogenous protective mechanism of lung injury by regulating HO-1 expression in septic mice.
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Objective To evaluate the role of mitophagy in hydrogen-induced reduction of lipopolysaccharide (LPS)-caused mitochondrial injury to macrophages of mice.Methods Macrophage line RAW264.7 cells of mice were routinely cultured and divided into 4 groups (n =6 each) using a random number table method:control group (Con group),LPS group,LPS plus hydrogen group (LPS + H2 group) and LPS plus hydrogen plus mitophagy inhibitor 3-methyladenine (3-MA) group (LPS+H2+3-MA group).Cells were incubated for 6 h with LPS at the concentration of 1 μg/ml in LPS group.Cells were incubated for 6 h with LPS 1 μg/ml and hydrogen-rich medium 0.6 mmol/L.Cells were incubated for 1 h with 2 mmol/L 3-MA and then incubated for 6 h with LPS 1 μg/ml and hydrogen-rich medium 0.6 mmol/L in LPS+H2+3-MA group.Mitochondrial respiratory control ratio (RCR) was measured using a Clark-type electrode.Mitochondrial membrane potential (MMP) was determined by JC-1 staining.Autophagosomes were counted with a transmission electron microscope.The expression of PTEN-induced putative kinase 1 (PINK1),E3 ubiquitin ligase (Parkin),microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ)and Beclin-1 was determined by Western blot.Results Compared with Con group,RCR and MMP were significantly decreased,the expression of PINK1,Parkin,LC3 Ⅱ and Beclin-1 was up-regulated,and the number of autophagosomes was increased in LPS group (P＜0.05).Compared with LPS group,RCR and MMP were significantly increased,the expression of PINK1,Parkin,LC3 Ⅱ and Beclin-1 was up-regulated,and the number of autophagosomes was increased in LPS+H2group (P＜0.05).Compared with LPS+H2 group,RCR and MMP were significantly decreased,the expression of PINK1,Parkin,LC3 Ⅱ and Beclin-1 was down-regulated,and the number of autophagosomes was decreased in LPS + H2 + 3-MA group (P＜0.05).Conclusion Enhanced mitophagy is involved in hydrogen-induced reduction of LPS-caused mitochondirial injury to macrophages of mice.
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Objective:To investigate whether facial expression recognition correct rates were changed in stable patients with major depressive disorder.Methods:A cross-sectional study was performed in patients with major depressive disorder (n=80) (according to the DSM-IV) and matched normal controls by age, gender and education year (n=41) in this study.All subjects would recognize five different expression (anger, contempt, fear, happiness and sadness).These emotions were displayed on four different male and female computerized cartoon faces, and each expression was shown once in a static format and once in a dynamic format.The facial expression recognition correct percentage would be compared between the two groups.Results:The correct identification rate of contempt was lower in the patients than in the normal controls (93％ vs.94％, P＜0.05).The correct identification rate of dynamic contempt was lower in the patients than in the normal controls (93％ vs.95％, P＜0.05).The correct identification rate of fear was lower in the patients than in the normal controls (89％ vs.93％, P＜0.05).The correct identification rate of dynamic fear was lower in the patients than in the normal controls (90％ vs.94％, P＜0.05).Regarding error rates of misattribution, compared with normal controls, depressive patients over-identified fear to anger (3.7％vs.1.8％) and contempt (2.9％vs.0.9％), sadness to contempt (5.0％ vs.1.7％) and happiness (2.0％ vs.0.2％) (Ps＜0.05).Conclusion:Patients with MDD have significantly great difficulties identifying negative expressions.
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Objective:To explore the effect of transitional nursing on medicine-taking compliance and insight of first hospitalized female schizophrenic patients.Methods:Seventy female cases who met ICD-10 diagnostic criteria that were discharged after stable treatment were randomly divided into intervention group and control group, 35 cases for each group.The medicine-taking compliance of the two groups was evaluated by the Morisky medication adherence scale, and the treatment effect was evaluated by the Insight and Treatment Attitude Questionnaire (ITAQ).The follow-up period was two months after discharge with four assessment points.Results:The Morisky scores in intervention group at the end of 1 week[ (4.7±1.2) vs. (4.0±0.8) ], 2 weeks[ (5.1±1.2) vs. (3.8±1.1) ], 1 month[ (5.1±1.5) vs. (3.5±1.2) ]and 2 months[ (5.8±1.2) vs. (3.3±1.0) ]were higher than those in control group (F=52.74, P＜0.001).The ITAQ scores in intervention group at the end of 1 week[ (3.4±2.1) vs. (2.9±1.4) ], 2 weeks[ (5.5±3.2) vs. (3.7±2.5) ], 1 month[ (4.6±2.7) vs. (4.5±2.7) ]and 2 months[ (6.9±3.4) vs. (3.9±2.6) ]were also higher than those in control group (F=7.12, P=0.010).Conclusion:Transitional nursing can improve the medicine-taking compliance and rehabilitation effect of first hospitalized female schizophrenic patients.
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Objective@#To evaluate the effect of hydrogen on mitochondrial biosynthesis in the hippocampus of mice with sepsis-associated encephalopathy (SAE).@*Method@#Two hundred and twenty-four healthy clean-grade male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups (n=56 each) using a random number table method: sham operation group (group SH), sham operation plus hydrogen group (group SH+ H2), group SAE, and SAE plus hydrogen group (group SAE+ H2). The model of SAE was established by cecal ligation and puncture in anesthetized mice.Group SH+ H2 and group SAE+ H2 inhaled 2% hydrogen for 1 h starting from 1 and 6 h after operation, respectively.Twenty mice were selectde to record the postoperative 7-day survival rate.The remaining animals were sacrificed at 24 h after operation, and brain tissues were taken for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of neuronal apoptosis (by TUNEL), mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and ATP content (by a bioluminescence assay). The apoptosis rate was calculated.The expression of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) in hippocampus was determined by Western blot at 6, 12 and 24 h after operation.@*Results@#Compared with group SH, the postoperative 7-day survival rate was significantly decreased, the apoptosis rate of hippocampal neurons was increased, the contents of MMP and ATP were decreased, and the expression of PGC-1α was up-regulated in SAE and SAE+ H2groups (P<0.05), and no significant change was found in the parameters mentioned above in group SH+ H2(P>0.05). Compared with group SAE, the postoperative 7-day survival rate was significantly increased, and the apoptosis rate of hippocampal neurons was decreased, the contents of MMP and ATP were increased, and the expression of PGC-1α was up-regulated (P<0.05), and the pathological changes of hippocampal tissues were significantly attenuated in group SAE+ H2.@*Conclusion@#The mechanism by which hydrogen mitigates SAE may be related to promoting mitochondrial biosynthesis in mice.
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Objective@#To evaluate the effect of hydrogen on the mitochondrial function in brain tissues of mice with sepsis-associated encephalopathy (SAE).@*Methods@#Two hundred and sixty-eight healthy male C57 mice, aged 6 weeks, weighing 20-25 g, were divided into 4 groups (n=67 each) using a random number table method: sham operation group (group SH), sham operation plus hydrogen gas group (group SH+ H2), group SAE, and SAE plus hydrogen gas group (group SAE+ H2). Sepsis was produced by cecal ligation and puncture in anesthetized mice.The mice in group SH+ H2 and group SAE+ H2 inhaled 2% hydrogen gas for 1 h starting from 1 and 6 h after operation, respectively.The postoperative 7-day survival rate was recorded.Brain tissues were obtained at 24 h after operation to count the normal neurons in hippocampal CA1 region.At 6, 12 and 24 h after operation, hippocampal mitochondria were isolated for determination of mitochondrial membrane potential (MMP) by fluorescence spectrophotometry and ATP content by a bioluminescence assay.Y-maze (spontaneous alternation) test was performed at days 3, 5 and 7 after operation.@*Results@#Compared with group SH, the 7-day survival rate was significantly decreased, the number of normal neurons in hippocampal CA1 region, MMP, mitochondrial ATP content and spontaneous alternation percentage in Y-maze test were significantly decreased in group SAE and group SAE+ H2 (P<0.05). Compared with group SAE, the 7-day survival rate, the number of normal neurons in hippocampal CA1 region, MMP, mitochondrial ATP content and spontaneous alternation percentage in Y-maze test were significantly increased in group SAE+ H2 (P<0.05).@*Conclusion@#The mechanism by which hydrogen reduces SAE is probably associated with improving mitochondrial function in brain tissues of mice.
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Objective To evaluate the effect of hydrogen on the mitochondrial function in brain tis-sues of mice with sepsis-associated encephalopathy(SAE).Methods Two hundred and sixty-eight healthy male C57 mice,aged 6 weeks,weighing 20-25 g,were divided into 4 groups(n＝67 each)using a ran-dom number table method: sham operation group(group SH),sham operation plus hydrogen gas group(group SH+H2),group SAE,and SAE plus hydrogen gas group(group SAE+H2).Sepsis was produced by cecal ligation and puncture in anesthetized mice.The mice in group SH+H2 and group SAE+H2 inhaled 2％hydrogen gas for 1 h starting from 1 and 6 h after operation,respectively.The postoperative 7-day sur-vival rate was recorded.Brain tissues were obtained at 24 h after operation to count the normal neurons in hippocampal CA1 region.At 6,12 and 24 h after operation,hippocampal mitochondria were isolated for determination of mitochondrial membrane potential(MMP)by fluorescence spectrophotometry and ATP con-tent by a bioluminescence assay.Y-maze(spontaneous alternation)test was performed at days 3,5 and 7 after operation.Results Compared with group SH,the 7-day survival rate was significantly decreased,the number of normal neurons in hippocampal CA1 region,MMP,mitochondrial ATP content and sponta-neous alternation percentage in Y-maze test were significantly decreased in group SAE and group SAE+H2(P＜0.05).Compared with group SAE,the 7-day survival rate,the number of normal neurons in hipp-ocampal CA1 region,MMP,mitochondrial ATP content and spontaneous alternation percentage in Y-maze test were significantly increased in group SAE+H2(P＜0.05).Conclusion The mechanism by which hy-drogen reduces SAE is probably associated with improving mitochondrial function in brain tissues of mice.
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Objective To evaluate the effect of hydrogen on mitochondrial biosynthesis in the hippocampus of mice with sepsis-associated encephalopathy (SAE).Method Two hundred and twenty-four healthy clean-grade male C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n=56 each) using a random number table method:sham operation group (group SH),sham operation plus hydrogen group (group SH+H2),group SAE,and SAE plus hydrogen group (group SAE+H2).The model of SAE was established by cecal ligation and puncture in anesthetized mice.Group SH+H2 and group SAE+H2 inhaled 2％ hydrogen for 1 h starting from 1 and 6 h after operation,respectively.Twenty mice were selectde to record the postoperative 7-day survival rate.The remaining animals were sacrificed at 24 h after operation,and brain tissues were taken for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of neuronal apoptosis (by TUNEL),mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and ATP content (by a bioluminescence assay).The apoptosis rate was calculated.The expression of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) in hippocampus was determined by Western blot at 6,12 and 24 h after operation.Results Compared with group SH,the postoperative 7-day survival rate was significantly decreased,the apoptosis rate of hippocampal neurons was increased,the contents of MMP and ATP were decreased,and the expression of PGC-1α was up-regulated in SAE and SAE+H2groups (P＜0.05),and no significant change was found in the parameters mentioned above in group SH+H2 (P＞0.05).Compared with group SAE,the postoperative 7-day survival rate was significantly increased,and the apoptosis rate of hippocampal neurons was decreased,the contents of MMP and ATP were increased,and the expression of PGC-1α was up-regulated (P＜0.05),and the pathological changes of hippocampal tissues were significantly attenuated in group SAE+H2.Conclusion The mechanism by which hydrogen mitigates SAE may be related to promoting mitochondrial biosynthesis in mice.
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Objective To investigate the effects of autophagy on N-methy-D-aspartate (NMDA) receptor and its subunit NR2B and behavioral test in a rat model of neuropathic pain (NP). Methods Male Sprague-Dawley (SD) rats were divided into sham group, NP group, autophagy inhibitor 3-methyladenine (3-MA) pretreatment group (3-MA+NP group) and autophagy inducer rapamyein (Rap) group (Rap+NP group) by random number table with 22 rats in each group. NP animal model was reproduced by ligating sciatic nerve, while sciatic nerve of the rats in the sham group were only exposed but not ligated. The rats in two pretreatment groups were intraperitoneally challenged with 3-MA 15 mg/kg or Rap 10 mg/kg injection 1 hour before operation. Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before and 1, 3, 7, 14 days after operation in each group. Spinal cord tissues were harvested at 1 day and 7 days after operation for autophagosome observation by electron microscope. The expressions of autophagy protein microtubule-associated protein 1 light chain 3 -Ⅱ (LC3 -Ⅱ), Beclin1, and NMDA, NR2B were determined by Western Blot. The positive expression of LC3 was detected by immunofluorescence. Results Compared with sham group, the MWT and TWL of rats in NP group were decreased gradually with the prolongation of operation time, the number of autophagosome, the expressions of LC3 -Ⅱ, Beclin1, NMDA, NR2B, and the positive expression of LC3 in spinal cord were significantly increased at 1 day after operation and till 7 days, which indicated that NP led to hyperpathia and autophagy activation. Compared with NP group, MWT was significantly further decreased, TWL was further shortened, the number of autophagosome was decreased, the expressions of LC3 -Ⅱ and Beclin1 in spinal cord were decreased, and NMDA and NR2B expressions were further increased after 3-MA pretreatment, with significant differences at 1 day after operation [MWT (g): 29.4±2.4 vs. 42.5±6.6, TWL (s): 7.2±1.0 vs. 8.8±1.1, LC3 -Ⅱ/β-actin: 0.38±0.03 vs. 0.52±0.07, Beclin1/β-actin: 0.29±0.06 vs. 0.59±0.05, NMDA/β-actin: 0.62±0.06 vs. 0.50±0.06, NR2B/β-actin: 0.57±0.03 vs. 0.46±0.03, all P < 0.05]. Immunofluorescence staining confirmed that the positive expression of LC3 was significantly decreased. Rap pretreatment could increase MWT, TWL and the number of autophagosome, increase LC3 -Ⅱ and Beclin1 expressions in spinal cord, and decrease NMDA and NR2B expressions in NP rats, and significant differences at 1 day after operation were found as compared with those in NP group [MWT (g): 49.4±4.4 vs. 42.5±6.6, TWL (s): 10.5±1.2 vs. 8.8±1.1, LC3 -Ⅱ/β-actin: 0.67±0.09 vs. 0.52±0.07, Beclin1/β-actin: 0.71±0.08 vs. 0.59±0.05, NMDA/β-actin: 0.40±0.05 vs. 0.50±0.06, NR2B/β-actin: 0.34±0.04 vs. 0.46±0.03, all P < 0.05], and immunofluorescence showed that the positive expression of LC3 was increased and lasted for 7 days. It indicated that Rap could increase the activity of autophagy, alleviate the occurrence of hyperalgesia, and reduce the expressions of NMDA receptor and its NR2B subunit. Conclusion NP could regulate the variety of NMDA/NR2B and hyperalgesia via increasing autophagy.
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Objective To evaluate the effect of dexmedetomidine on expression of hypoxia-inducible factor-1α (HIF-1α) during endotoxin-caused apoptosis in macrophages of mice.Methods Mouse macrophage cell line RAW264.7 cultured in vitro were seeded in 6-well or 96-well plates and divided into 4 groups (n=16 each) when cell confluence reached 60％-70％ using a random number table method:control group (group Con),dexmedetomidine group (group Dex),lipopolysaccharide (LPS) group,and LPS plus dexmedetomidine group (group LPS+Dex).Phosphate buffer solution was added in group Con.Dexmedetomidine 1 μmol/L was added in group Dex.LPS 1 μg/ml was added in LPS and LPS+Dex groups.Dexmedetomidine 1 μmol/L was added immediately after adding LPS in group LPS+Dex.Cells were then cultured for 24 h in each group.Cell apoptosis was measured using TUNEL,mitochondrial membrane potential using JC-1,reactive oxygen species (ROS) content by ROS kit,and ATP content by ATP kit.The apoptosis rate was calculated.The expression of HIF-1α,cytochrome C (Cyt-c),caspase-9 and cleaved caspase-3 was detected by Western blot.Results Compared with group Con,the apoptosis rate and ROS content were significantly increased,ATP content and mitochondrial membrane potential were decreased,the expression of HIF-1α,Cyt-c,caspase-9 and cleaved caspase-3 was up-regulated in group LPS (P＜ 0.05),and no significant change was found in the parameters mentioned above in group Dex (P＞0.05).Compared with group LPS,the apoptosis rate and ROS content were significantly decreased,ATP content and mitochondrial membrane potential were increased,the expression of HIF-1α was up-regulated,and the expression of Cyt-c,caspase-9 and cleaved caspase-3 was down-regulated in group LPS + Dex (P＜0.05).Conclusion Dexmedetomidine can reduce endotoxin-caused oxidative stress injury to macrophages,improve mitochondrial function and inhibit mitochondrial apoptosis,and the mechanism may be related to upregulating the expression of HIF-1α in mice.
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Objective:To compare the cognitive functions in patients with depressive disorder,hyperlipidemia disorder,and comorbid both of the disorders.Methods:A cross-sectional study was performed in age,gender and education year matched patients with depressive disorder (n =51)(according to the ICD-10),hypedipidemia(n =38) (according to the Chinese adult lipid guideline),comorbid both of the disorders(n =40) and normal controls (n =56) were recruited in this study.All subjects received a battery of neuropsychological tests to access the anxiety and depression symptoms and cognitive function.Results:The scores of MoCA were lower in the patients with comorbid both disorders and patients with depression than patients with hypedipidemia [(24 ± 3),(24 ± 4)vs.(26 ± 3),Ps ＜0.05],and were lower in patients with depression than in normal controls(25 ±3),P ＜0.05.Stroop color test amends numbers were higher in patients with comorbid disorder than in the other three groups (Ps ＜0.05).The scores of immediate and delayed logical memory were higher in patients with hyperlipidemia than in other three groups (Ps ＜0.05).The total number of words in verbal fluency test were lower in patients with comorbid disorders and patients with depression than in patients with hyperlipidemia (Ps ＜0.05).Wisconsin card sorting test category completes were lower in patients with comorbid disorders and patients with depression than in patients with hyperlipidemia and normal controls (Ps ＜ 0.05).The scores of persistent errors were higher in patients with comorbid disorders and patients with depression than in patients with hyperlipidemia and normal controls (Ps ＜0.05).Conclusion:In this study,patients with depressive disorder have impairment of cognitive function,while hyperlipidemia may probably do not impair cognitive function.
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Objective:To investigate the natural course and psychosocial risk factors of depressive symptoms in college freshmen.Methods:Changes in depressive symptoms among college freshmen were observed for two year periods,self-reporting questionnaires were used to collect data every half yearly.The depressive symptoms were assessed with the Centre for Epidemiological Study-Depression Scale,and psychosocial factors were tested as the potential risk factors,rating by Zung Self-Rating Anxiety Scale,Eysenck Personality Questionnaire-Neuroticism Subscale,Rosenberg Self-Esteem Scale,Frost Multidimensional Perfectionism Scale,Parental Rearing Scale,Adolescents Self-Rating Life Events Checklist,Epworth Sleepiness Scale,daytime sleep time,Godin Leisure-Time Exercise Questionnaire.Generalized estimating equations model was used to analyze the date.Results:Totally 1339 college students were recruited in the survey.It was found the depressive scores significantly increased in the second and third surveys,then decreased in the fourth and fifth surveys.Male (Coef=-1.01,SE =0.42,P =0.017),not interested in the major (Coef=3.89,SE =1.42,P =0.006),neuroticism(Coef =0.79,SE =0.23,P =0.001),self-esteem(Coef =-1.57,SE =0.25,P ＜ 0.001),anxiety(Coef =4.79,SE =0.16,P ＜ 0.001),life events (Coef =0.08,SE =0.01,P ＜ 0.00l),daytime sleepiness (Coef =0.80,SE =4.56,P ＜ 0.001) were significantly correlated with depressive symptoms.Conclusion:The first year of college is a critical time in the prevention depression.Not interested in the major,high neuroticism,low self-esteem,anxiety,life events,daytime sleepiness may be risk factors of depressive symptom in college students.
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Objective To evaluate the role of spinal nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in hydrogen-induced reduction of inflammatory pain in rats.Methods Sixty-four SPF healthy adult male Sprague-Dawley rats,weighing 200-250 g,were divided into 4 groups (n =16 each) using a random number table:control group (group C),inflammatory pain group (group IP),inflammatory pain plus hydrogen-rich saline group (group IP+H2) and inflammatory pain plus hydrogen-rich saline plus Nrf2 inhibitor all-trans retinoic acid (ATRA) group (group IP+H2+ATRA).Chronic inflammatory pain was induced by injecting complete Freund's adjuvant (CFA) 100 μl into the plantar surface of the left hind paw in IP group and IP+H2 group.The equal volume of normal saline was given instead in group C.Hydrogen-rich saline 5 ml/kg was injected intraperitoneally once a day for 7consecutive days starting from 1 day after injecting CFA in group IP+H2 and group IP+H2+ATRA,and the equal volume of normal saline was given instead in the other groups.ATRA 7 mg/kg was injected intraperitoneally once a day for 2 consecutive days starting from 2 days before injecting CFA.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before establishing the model (T0) and 1,3 and 7 days after establishing the model (T1-3).Six rats were sacrificed after the last measurement of pain threshold on day 7 after establishing the model,and the L4-6 lumbar segments of the spinal cord were removed for determination of the expression of Nrf2,HO-1 and glial fibrillary acidic protein (GFAP) by Western blot.Results Compared with group C,the MWT was significantly decreased and the TWL was shortened at T1-3,and the expression of Nrf2,HO-1 and GFAP was up-regulated in IP and IP+H2 groups (P＜0.05).Compared with group IP,the MWT was significantly increased and the TWL was prolonged at T1-3,the expression of Nrf2 and HO-1 was up-regulated,and the expression of GFAP was down-regulated in group IP+H2 (P＜0.05),and no significant change was found in the parameters mentioned above in group IP+H2+ATRA (P＞0.05).Compared with group IP+H2,the MWT was significantly decreased and the TWL was shortened at T1-3,the expression of Nrf2 and HO-1 was down-regulated,and the expression of GFAP was up-regulated in group IP+H2+ATRA (P＜0.05).Conclusion Activation of spinal Nrf2/HO-1 signaling pathway is involved in hydrogen-induced reduction of infflammatory pain in rats.
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Objective To evaluate the relationship between phosphatidylinositol 3?kinase∕serine?threonine kinase(PI3K∕Akt)signaling pathway and autophagy during acute lung injury in septic mice. Methods Thirty?six male C57BL∕6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups(n=12 each)using a random number table: sham operation group(group SH), sepsis group (group S)and PI3K inhibitor LY294002 plus sepsis group(group LY+S). Sepsis was induced by cecal ligation and puncture in S and LY+S groups. LY294002 30 mg∕kg was intraperitoneally injected at 2 h be?fore operation in group LY+S. Arterial blood samples were taken at 24 h after operation for blood gas analy?sis, PaO2was recorded, and oxygenation index was calculated. Lung specimens were obtained for examina?tion of pathological changes which were scored and for determination of autophagosome count(using trans?mission electron microscope), wet∕dry weight ratio(W∕D ratio)and expression of Akt, phosphorylated Akt(p?Akt), Beclin?1 and microtubule?associated protein 1 light chain 3 Ⅱ(LC3 Ⅱ). The p?Akt∕Akt ratio was calculated. Results Compared with group SH, oxygenation index was significantly decreased,and the W∕D ratio and pathological score were increased in S and LY+S groups, the autophagosome count was significantly increased, p?Akt∕Akt ratio was increased, and the expression of Beclin?1 and LC3Ⅱ was up?regulated in group S(P<0.05). Compared with group S, oxygenation index was significantly de?creased, and the W∕D ratio was increased, the autophagosome count was decreased, pathological scores were increased, p?Akt∕Akt ratio was decreased, and the expression of Beclin?1 and LC3Ⅱwas down?regu?lated in group LY+S(P<0.05). Conclusion PI3K∕Akt signaling pathway activation mediates autophagy and is involved in the endogenous protective mechanism of acute lung injury in septic mice.
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Objective To investigate the role of nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome in hydrogen-rich saline-induced reduction of lipopolysaccharide (LPS)-caused damage to mitochondria in macrophages of mice.Methods Macrophage line RAW264.7 of mice were routinely cultured and divided into 4 groups (n=6 each) using a random number table method:control group (group C),group LPS,hydrogen-rich saline plus LPS group (group LPS+H2) and hydrogen-rich saline plus LPS plus ATP group (group LPS+ATP+H2).LPS was given at the concentration of 1 μg/ml,and the cells were then incubated for 30 min in group LPS.LPS at the concentration of 1 μg/ml and hydrogen-rich saline at the concentration of 0.6 mmol/L were simultaneously given,and the cells were then incubated for 30 min in LPS+H2 and LPS+ATP+H2 groups.ATP at the concentration of 1 nmol/L was then given,and the cells were incubated for 6 h in group LPS+ATP+H2.Mitochondrial membrane potential (MMP) was determined by JC-1 staining,and respiratory control ratio (RCR) was measured using a Clark-type electrode.The expression of NLRP3,caspase-1 and apoptosisassociated speck-like protein containing C-terminal caspase recruitment domain (ASC) was determined by Western blot.The concentrations of INTERLEUKIN-1 BETA (IL-1β),IL-18 and IL-6 in the supernatant were determined by enzyme-linked immunosorbent assay.Results Compared with group C,MMP and RCR were significantly decreased,the concentrations of IL-1β,IL-18 and IL-6 in the supernatant were increased,and the expression of NLRP3,caspase-1 and ASC was up-regulated in group LPS (P＜0.05).Compared with group LPS,MMP and RCR were significantly increased,the concentrations of IL-1β,IL-l8 and IL-6 in the supernatant were decreased,and the expression of NLRP3,caspase-1 and ASC was down-regulated in group LPS+H2 (P＜0.05).Compared with group LPS+H2,MMP and RCR were significantly increased,the concentrations of IL-1β,IL-18 and IL-6 in the supernatant were decreased,and the expression of NLRP3,caspase-1 and ASC was down-regulated in group LPS+ATP+H2 (P＜0.05).Conclusion Hydrogen-rich saline can reduce LPS-caused damage to mitochondria in macrophages of mice through inhibiting the activation of NLRP3 inflammasome.
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Objective To evaluate the role of autophagy in dexmedetomidine-induced reduction of lipopolysaccharide ( LPS)-caused inflammatory responses in macrophages of mice. Methods Mouse mac-rophage cell line RAW264. 7 cultured in vitro were seeded in 6-well or 96-well plates and divided into 4 groups ( n=20 each ) when cell confluence reached 60％ using a random number table method: control group (group Con), LPS group, LPS plus dexmedetomidine group (group LPS+DEX), and LPS plus dexmedetomidine plus autophagy inhibitor 3-MA group (group LPS+DEX+3-MA). PBS was added and cells were cultured for 12 h in group Con. LPS at the final concentration of 1000 ng∕ml was added and cells were incubated for 12 h in group LPS. LPS at the final concentration of 1000 ng∕ml was added, and then dexmedetomidine at the final concentration of 1 μmol∕L was immediately added, and cells were incubated for 12 h in group LPS+Dex. In group LPS+Dex+3-MA, 3-MA at the final concentration of 2 mmol∕L was added and cells were incubated for 1 h, LPS at the final concentration of 1000 ng∕ml was added, and then dexmedetomidine at the final concentration of 1 μmol∕L was immediately added, and cells were incubated for 12 h. Cell viability was detected by CCK-8 assay, and the concentrations of nitrous oxide ( NO) , tumor necrosis factor-alpha ( TNF-α) and interleukin-1beta ( IL-1β) in the supernatant were determined by en-zyme-linked immunosorbent assay, and the expression of microtubule-associated protein 1 light chain 3 Ⅰ( LC3 Ⅰ) , LC3Ⅱ, P62 and Bcelin-1 was detected by Western blot. LC3Ⅱ∕LC3Ⅰ ratio was calculated. Results Compared with group Con, the cell viability was significantly decreased, the concentrations of TNF-α, IL-1β and NO and LC3Ⅱ∕LC3Ⅰratio were increased, and the expression of P62 and Beclin1 was up-regulated in group LPS (P<0. 05). Compared with group LPS, the cell viability was significantly in-creased, the concentrations of TNF-α, IL-1βand NO were decreased, LC3Ⅱ∕LC3Ⅰratio was increased, the expression of P62 was down-regulated, and the expression of Beclin1 was up-regulated in group LPS+DEX ( P<0. 05) . Compared with group LPS+Dex, the cell viability was significantly decreased, the con-centrations of TNF-α, IL-1β and NO were increased, LC3Ⅱ∕LC3Ⅰ ratio was decreased, the expression of P62 was up-regulated, and the expression of Beclin1 was down-regulated in group LPS+Dex+3-MA ( P<0. 05) . Conclusion Enhanced autophagy is involved in dexmedetomidine-induced reduction of LPS-caused inflammatory responses in macrophages of mice.
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Objective To evaluate the effect of hydrogen on mitochondrial dynamics during endotoxin-induced damage to human umbilical vein endothelial cells (HUVECs).Methods HUVECs cultured in vitro were seeded in the culture plate and divided into 4 groups using a random number table:control group (group C),hydrogen-saturated culture medium group (H group),endotoxin group (group E) and endotoxin + hydrogen-saturated culture medium group (group E+H).The cells were cultured in the plain culture medium in C and E groups.The cells were cultured in the hydrogen-saturated culture medium containing lipopolysaccharide (LPS) with the final concentration of 10 μg/ml in H and E+H groups.At 2,8 and 24 h of culture or incubation with LPS,the cell viability was detected by methyl thiazolyl tetrazolium assay,the intracellular ATP content was measured using the phosphomolybdic acid colorimetric method,and the expression of dynamin-related protein 1 (DRP1) was detected by using Western blot.The expression of DRP1 was detected by immunofluorescence at 8 h of incubation with LPS.Results Compared with group C,the cell viability and ATP content were significantly decreased,and the expression of DRP1 was up-regulated at each incubation time point in E and E +H groups (P＜0.05),and no significant change was found in the parameters mentioned above in group H (P＞0.05).Compared with group E,the cell viability and ATP content were significantly increased,and the expression of DRP1 was down-regulated at each incubation time point in group E+H (P＜0.05).Conclusion The mechanism by which hydrogen reduces endotoxin-induced damage to HUVECs is related to down-regulation of DRP1 expression and inhibition of excessive mitochondrial fission.