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Objective:To evaluate the role of the autophagy in hydrogen-induced reduction of myocardial injury in septic mice.Methods:A total of 192 clean-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 6 groups ( n=32 each) using a random number table method: sham operation group (group Sham), sham operation plus hydrogen group (group Sham+ H 2), sepsis group (group S), sepsis plus hydrogen group (group S+ H 2), sepsis plus bafilomycin A1 group (group S+ BafA1) and sepsis plus hydrogen plus bafilomycin A1 group (group S+ H 2+ BafA1). Sepsis was produced by cecal ligation and puncture (CLP) after anesthesia. The mice inhaled 2% hydrogen for 1 h starting from 1 and 6 h after operation in group Sham+ H 2, group S+ H 2 and group S+ H 2+ BafA1. Bafilomycin A1 1 mg/kg was intraperitoneally injected at 1 h after operation in S+ BafA1 and S+ H 2+ BafA1 groups. Twenty mice in each group were selected to record the 7-day survival rates after operation. Then the mice were sacrificed at 24 h after operation to observe the pathological changes of myocardial tissues which were scored and detect the serum cardiac troponin I (cTnI) concentration (by enzyme-linked immunosorbent assay) and determine the level of microtubule-associated protein 1 light chain 3 B (LC3B) and P62 (by Western blot). LC3Ⅱ/LC3Ⅰratio was calculated. Results:Compared with group Sham, the 7-day survival rate after operation was significantly decreased, the serum cTnI concentrations and pathological scores of myocardial tissues were increased, the expression of P62 was up-regulated ( P<0.05), no significant change was found in LC3Ⅱ/LC3Ⅰratio ( P>0.05), and no significant change was found in the parameters mentioned above in group Sham+ H 2 ( P>0.05). Compared with group S, the 7-day survival rate after operation was significantly increased, the serum cTnI concentrations and pathological scores of myocardial tissues were decreased, LC3Ⅱ/LC3Ⅰratio was increased, and the expression of P62 was down-regulated in group S+ H 2, and LC3Ⅱ/LC3Ⅰratio was significantly decreased, and the expression of P62 was up-regulated in group S+ BafA1 ( P<0.05). Compared with group S+ H 2, the 7-day survival rate was significantly decreased, the serum cTnI concentrations and pathological scores of myocardial tissues were increased, LC3Ⅱ/LC3Ⅰratio was decreased, and the expression of P62 was up-regulated in group Sham+ H 2 ( P<0.05). Conclusions:The mechanism by which hydrogen alleviates myocardial damage may be related to promoting autophagy in septic mice.
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Objective:To evaluate the effect of hydrogen-rich saline (HRS) on mitochondrial biogenesis and dynamics in hippocampus of mice with sepsis-associated encephalopathy (SAE).Methods:One hundred and twenty-eight male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=32 each) using a random number table method: sham operation group (Sham group), sham operation plus HRS group (Sham+ HRS group), SAE group and SAE plus HRS group.Sepsis was developed by cecal ligation and puncture (CLP) in anesthetized mice.HRS 10 ml/kg was intraperitoneally injected at 1 and 6 h after CLP in Sham+ HRS and SAE+ HRS groups.Twenty mice were randomly selected from each group to record the 7-day survival after operation.The working memory of the mice was observed by Y-maze test on days 3, 5 and 7 after CLP.The hippocampal tissues were obtained at 24 h after CLP for determination of the content of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) (by enzyme-linked immunosorbent assay), activities of superoxide dismutase (SOD) and catalase (CAT) (by spectrophotometry), and expression of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α), nuclear respiratory factor 2 (NRF2), mitochondrial transcription factor A (Tfam), dynamin-related protein 1 (Drp1) and mitochondrial fusion protein mitofusin 2 (Mfn2) (by Western blot). Results:Compared with group Sham, the postoperative 7-day survival rate was significantly decreased, the time spent in novel arm was shortened, the contents of TNF-α, IL-6 and HMGB1 were increased, the activities of SOD and CAT were decreased, the expression of PGC-1α, NRF2 and Tfam was up-regulated, the expression of Drp1 was up-regulated, and the expression of Mfn2 was down-regulated in group SAE ( P<0.05). Compared with group SAE, the postoperative 7-day survival rate was significantly increased, the time spent in novel arm was prolonged, the contents of TNF-α, IL-6 and HMGB1 were decreased, the activities of SOD and CAT were increased, the expression of PGC-1α, NRF2 and Tfam was up-regulated, the expression of Drp1 was down-regulated, and the expression of Mfn2 was up-regulated in group SAE+ HRS ( P<0.05). Conclusions:The mechanism by which HRS alleviates SAE may be related to promotion of mitochondrial biogenesis, regulation of dynamics, and reduction of oxidative stress in hippocampus of mice.
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Objective:To evaluate the effect of hydrogen on mitochondrial dynamics in hippocampus of mice with sepsis-associated encephalopathy (SAE).Methods:A total of 224 clean-grade healthy male C57 mice, weighing 20-25g, aged 6-8 weeks, were divided into 4 groups ( n=56 each) using a random number table method: sham operation group (group Sham), sham operation + hydrogen group (group Sham+ H 2), SAE group and SAE + hydrogen group (group SAE+ H 2). Sepsis was produced by cecum ligation and puncture (CLP). Sham+ H 2and SAE+ H 2 groups inhaled 2% hydrogen for 1 h starting from 1 and 6 h after CLP, respectively.The postoperative 7-day survival rate was recorded.Brain tissues were obtained at 24 h after operation for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and ATP content (by a bioluminescence assay) in hippocampal tissues.At 6, 12 and 24 h after operation, hippocampal mitochondria were isolated for determination of the expression of dynamin-related protein 1 (Drp1) and the mitochondrial fusion proteins mitofusin 2 (Mfn2) (by Western blot). Results:Compared with group Sham, the postoperative 7-day survival rate was significantly decreased, the contents of MMP and ATP were decreased, the expression of Drp1 was up-regulated, and the expression of Mfn2 was down-regulated( P<0.05), the pathological changes were aggravated in hippocampal CA1 region in SAE and SAE+ H 2 groups, and no significant change was found in the parameters mentioned above in group Sham+ H 2 ( P>0.05). Compared with group SAE, the postoperative 7-day survival rate was significantly increased, the contents of MMP and ATP were increased, the expression of Drp1 was down-regulated, and the expression of Mfn2 was up-regulated( P<0.05), the pathological changes were attenuated in hippocampal CA1 region in group SAE+ H 2. Conclusion:The mechanism by which hydrogen improves mitochondrial function is probably associated with promoting mitochondrial fusion and inhibiting mitochondrial fission in hippocampus of mice with SAE.
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Objective:To evaluate the effect of hydrogen on activation of A1 astrocytes in the hippocampus of mice with sepsis-associated encephalopathy (SAE).Methods:A total of 164 clean-grade healthy male C57BL/6J mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=41 each) using a random number table method: sham operation group (group Sham), sham operation plus hydrogen group (group Sham+ H 2), group SAE and SAE plus hydrogen group (group SAE+ H 2). The SAE model was established by cecal ligation and perforation.Group Sham+ H 2 and group SAE+ H 2 inhaled 2% hydrogen starting from 1 and 6 h after operation, respectively.Twenty mice in each group were selected to observe the 7-day survival rate after operation.The remaining mice were sacrificed at 12 h after operation, and brain tissues were removed for examination of the pathological changes in hippocampal CA1 region (with a light microscope) and for determination of the apoptosis in neurons (by TUNEL), co-expression of hippocampal glial fibrillary acidic protein (GFAP) and complement C3 (by immunofluorescence staining), expression of A1 astrocyte marker C3 (by Western blot), and contents of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) (by enzyme-linked immunosorbent assay). The abnormal cell ratio and apoptosis rate were calculated.Six mice in each group were selected at 7 days after operation to perform Y-Maze paradigm. Results:Compared with group Sham, the 7-day survival rate after operation was significantly decreased, the abnormal cell ratio and apoptosis rate of hippocampal neurons were increased, the contents of TNF-α, IL-6 and HMGB1 were increased, the expression of C3 was up-regulated, the number of cells coexpressing GFAP and C3 was increased, the exploration time spent in the novel arm in Y-Maze paradigm was shortened, and the preference index was decreased in group SAE ( P<0.05). Compared with group SAE, the 7-day survival rate after operation was significantly increased, the abnormal cell ratio and apoptosis rate of hippocampal neurons were decreased, the contents of TNF-α, IL-6 and HMGB1 were decreased, the expression of C3 was down-regulated, the number of cells coexpressing GFAP and C3 was decreased, the exploration time spent in the novel arm in Y-Maze paradigm was prolonged, and the preference index was increased in group SAE+ H 2 ( P<0.05). There was no significant difference in each parameter mentioned above between Sham group and Sham+ H 2 group ( P>0.05). Conclusion:The mechanism by which hydrogen improves SAE may be related to inhibiting activation of A1 type astrocytes in mice.
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Objective:To evaluate the relationship between phosphorylation of Tau protein and apolipoprotein E (ApoE) containing 18 kDa fragments and investigate the mechanism of neuronal damage induced by sevoflurane.Methods:Primary neurons (ApoE3 and ApoE2 genotypes, 24 dishes for each genotype) of fetal mice cultured until the 5th day were divided into 4 groups ( n=12 each) using a random number table method: ApoE3 control group (A3C group), ApoE3 sevoflurane group (A3S group), ApoE2 control group (A2C group) and ApoE2 sevoflurane group (A2S group). Neurons were treated with 21% oxygen + 5% carbon dioxide + 4.1% sevoflurane for 4 h in A3S and A2S groups, while the neurons were only treated with 21% oxygen + 5% carbon dioxide in A3C and A2C groups.The cell proteins were then extracted to detect the expression of full-length ApoE and ApoE, AT8 and PHF1 containing 18 kDa fragments (by Western blot), expression of ApoE mRNA (by real-time polymerase chain reaction), and concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in the supernatant (by enzyme-linked immunosorbent assay). Results:Compared with A2C group, the expression of ApoE mRNA and full-length ApoE in neurons was up-regulated ( P<0.05), and no significant change was found in the expression of AT8 and PHF1 and concentrations of TNF-α and IL-6 in the supernatant in A2S group ( P>0.05). Compared with A3C group, the expression of ApoE mRNA, full-length ApoE, and ApoE, AT8 and PHF1 containing 18 kDa fragments was up-regulated, and the concentrations of TNF-α and IL-6 in the supernatant were increased in A3S group ( P<0.05). Conclusion:Sevoflurane may promote phosphorylation of Tau proteins and increase inflammatory responses through up-regulating the expression of ApoE containing 18 kDa fragments, thus leading to neuronal damage.
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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 dexmedetomidine on intestinal injury in severely burned rats.Methods:One hundred and twenty healthy clean-grade male Sprague-Dawley rats, weighing 240-260 g, were divided into 4 groups ( n=30 each) using a random number table method: sham operation group (group Sham), sham plus dexmedetomidine group (group Sham+ Dex), severe burn group (group Burn), and severe burn plus dexmedetomidine group (group Burn+ Dex). Forty percent total body surface area of III degree burn model was developed in chloral hydrate-anesthetized rats.Dexmedetomidine was intravenously infused for 4 h at a rate of 5 μg·kg -1·h -1 starting from 3 h after establishing the model in Sham+ Dex group and Burn+ Dex group.The small intestinal tissues were removed for examination of the pathological changes which were scored and for determination of tumor necrosis factor-alpha (TNF-α) and high-mobility group box 1 protein (HMGB1) contents (by enzyme-linked immunosorbent assay), and expression of occludin and ZO-1 protein (by Western blot). The serum concentrations of 4-kD-FITC were measured at 90, 180, 360 and 720 min after establishing the model. Results:Compared with Sham group, the pathological scores of intestinal tissues, contents of TNF-α and HMGB1, serum concentrations of 4-kD-FITC at each time point were significantly increased, and the expression of occludin and ZO-1 was down-regulated in Burn group and Burn+ Dex group ( P<0.05), and no significant change was found in the parameters mentioned above in Sham+ Dex group ( P>0.05). Compared with Burn group, the pathological scores of intestinal tissues, contents of TNF-α and HMGB1, serum concentrations of 4-kD-FITC at each time point were significantly decreased, and the expression of occludin and ZO-1 was up-regulated in Burn+ Dex group ( P<0.05). Conclusion:Dexmedetomidine can reduce intestinal injury in severely burned rats, and the mechanism may be related to inhibiting inflammatory responses in the intestine.
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Objective:To evaluate the effect of hydrogen on lipopolysaccharide (LPS)-caused inflammatory responses in BV-2 microglia and the role of autophagy.Methods:The BV-2 microglial cells cultured in vitro were seeded in 6- or 96-well plates and were divided into 4 groups ( n=24 each) using a random number table method: control group (group C), group LPS, hydrogen-rich medium group (group H) and autophagy inhibitor 3-methylpurine group (group 3-MA). In group C, cells were cultured in MEM culture medium supplemented with 15% fetal bovine serum for 24 h. In group LPS, LPS was added at a final concentration of 1 μg/ml, and cells were incubated for 24 h. In group H, LPS was added at a final concentration of 1 μg/ml, the culture medium was replaced with a hydrogen-rich medium at a final concentration of 0.6 mmol/L, and cells were incubated for 24 h. In group 3-MA, 3-methylpurine was added at a final concentration of 2 mmol/L, and the subsequent treatment was similar to those previously described in group H. The cell survival rate was detected by CCK-8 assay.The concentrations of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10 and transforming growth factor-β (TGF-β) in supernatant were detected by enzyme-linked immunosorbent assay.The percentage of ionized calcium binding adaptor molecule-1 (Iba-1) +, Iba-1 + CD86 + and Iba-1 + CD206 + cells was detected by flow cytometry.The expression of microtubule-associated protein 1 light chain 3 Ⅰ (LC3 Ⅰ), LC3Ⅱ, Beclin-1 and p62 was detected by Western blot, and the ratio of LC3Ⅱ/LC3Ⅰ was calculated. Results:There was no significant difference in the cell survival rate among the four groups ( P>0.05). Compared with group C, the concentrations of TNF-α, IL-6, IL-10 and TGF-β and percentage of Iba-1 +, Iba-1 + CD86 + and Iba-1 + CD206 + cells were significantly increased in LPS, H and 3-MA groups, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was significantly down-regulated, and p62 expression was up-regulated in LPS and 3-MA groups, and the ratio of LC3LC3Ⅱ/LC3Ⅰ and Beclin-1 expression was significantly up-regulated, and p62 expression was down-regulated in group H ( P<0.05). Compared with group LPS, the concentrations of TNF-α and IL-6 were significantly decreased, the concentrations of IL-10 and TGF-β were increased, the percentage of Iba-1 + and Iba-1 + CD86 + cells were decreased, the percentage of Iba-1 + CD206 + cells was increased, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was up-regulated, and p62 expression was down-regulated in group H ( P<0.05), and no significant change was found in the above indexes in group 3-MA ( P>0.05). Compared with group H, the concentrations of TNF-α and IL-6 were significantly increased, the concentrations of IL-10 and TGF-β were decreased, the percentage of Iba-1 + and Iba-1 + CD86 + cells was increased, the percentage of Iba-1 + CD206 + cells was decreased, the LC3Ⅱ/LC3Ⅰ ratio and Beclin-1 expression was down-regulated, and p62 expression was up-regulated in group 3-MA ( P<0.05). Conclusion:The mechanism by which hydrogen reduces LPS-caused inflammatory responses in BV-2 microglia is related to enhancing autophagy and inhibiting microglial activation.
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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 evaluate the effect of hydrogen on the expression of matrix metalloproteinase-9 (MMP-9) in brain tissues of mice with sepsis-associated encephalopathy (SAE).Methods A total of 212 clean-grade healthy male C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =53 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H2),group SAE and SAE plus hydrogen group (group SAE+ H2).The sepsis model was established by intraperitoneally injecting human fecal suspension.Sham+H2 and SAE+H2 groups inhaled 2％ hydrogen for 1 h at 1 and 6 h after establishing the model,respectively.Postoperative 14-day survival rate was recorded.Evans blue (EB) was injected into the tail vein at 24 h after establishing the model,and the content of EB in brain tissues was calculated.Brain tissues were taken for determination of brain water content.The expression of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and high-mobility group box 1 protein (HMGB1) in hippocampal tissues was detected by enzymelinked immunosorbent assay.The expression of MMP-9 and ZO-1 protein and mRNA in hippocampal tissues was determined by Western blot and real-time polymerase chain reaction,respectively,at 6,12 and 24 h after establishing the model.Results Compared with Sham group,the 14-day survival rate was significantly decreased,the EB content in brain tissues,brain water content,and contents of TNF-α,IL-6 and HMGB1 were significantly increased,the expression of MMP-9 protein and mRNA was up-regulated,and the expression of ZO-1 protein and mRNA was down-regulated in SAE and SAE+H2 groups (P＜0.05),and no significant change was found in the parameters mentioned above in group Sham+H2 (P＞0.05).Compared with group SAE,the 14-day survival rate was significantly increased,the EB content in brain tissues,brain water content,and contents of TNF-α,IL-6 and HMGB1 were significantly decreased,the expression of MMP-9 protein and mRNA was down-regulated,and the expression of ZO-1 protein and mRNA was up-regulated in group SAE+H2 (P＜0.05).Conclusion The mechanism by which hydrogen attenuates the blood-brain barrier damage may be related to inhibiting MMP-9 expression and to reducing inflammatory responses in brain tissues of SAE mice.
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Objective T o observe the changes of cystathionine β-synthase (C B S ) expression in the cere-bral cortex after brain contusion at different tim es. Methods A n experim ental m odel of traum atic brain injury (T B I) in m ice w as established by an im proved w eight-drop device. T hen W estern blotting and im m unohistochem ical exam ination w ere used to detect the C B S expression in cerebral cortex around in-jury at different tim e points (1 h, 6 h, 12 h, 1 d, 2 d, 3 d, 7 d). Results T he results of W estern blotting revealed that the expression level of C B S w as dow n-regulated and reached its low est level at the 3rd days after injury, and then restored to norm al level after 7 days. T he results of im m unohistochem istry show ed that C B S w as present in the norm al brain cortex. C B S expression gradually decreased at the 3rd days after injury, and then restored to norm al level after 7 days. Conclusion C B S has the potential to be a reference index for tim e estim ation after brain contusion in forensic practice.
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The zinc finger protein A20 exists in many kinds of ceils in the body and plays multiple roles in physiological and pathological processes such as regulation of immune response,inflammatory diseases and cancers by inhibiting cell apoptosis induced by tumor necrosis factor (TNF) and restricting NFKB signaling pathway.Recently,the role of A20 in generation and development of tumors draws wide attention,hence,we summarized recent research progress of the relation between A20 and cancer in this review.
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Objective To evaluate the effect of dexmedetomidine on myocardial injury induced by renal ischemia-reperfusion (I/R) injury in rats.Methods Twenty-four male Wistar rats,weighing 280-300 g,were randomly divided into 3 groups (n =8 each):sham operation group (group S),group I/R and dexmedetomidine group (group Dex).Renal ischemia was induced by occlusion of bilateral renal arteries for 45 min followed by reperfusion in I/R and Dex groups.At 20 min before ischemia,dexmedetomidine 50 μg/kg was injected intraperitoneally in group Dex,and the rest procedures were similar to those previously described in group I/R.The rats were sacrificed at 24 h of reperfusion and myocardial specimens were obtained for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity.The apoptosis in cardiomyocytes was examined by flow cytometry.Apoptosis index was calculated.The expression of Bcl-2 and Bax was detected by immunohistochemistry,and Bcl-2/Bax ratio was calculated.Results Compared with group S,apoptosis index and MDA content were significantly increased in I/R and Dex groups,Bcl-2/Bax ratio was decreased in group I/R,and Bcl-2/Bax ratio was increased in group Dex.Compared with group I/R,apoptosis index and MDA content were significantly decreased,and SOD activity and Bcl-2/Bax ratio were increased in group Dex.Conclusion Dexmedetomidine can attenuate myocardial injury induced by renal I/R in rats,and the mechanism may be related to inhibited apoptosis in cardiomyocytes and reduced lipid peroxidation.
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Objective To evaluate the effects of sevoflurane preconditioning on lung injury induced by renal ischemia-reperfusion (I/R) in rats.Methods Forty adult male Wistar rats,aged 3 months,weighing 200-250 g,were randomly divided into 5 groups (n =8 each):sham operation group (group S); renal I/R group (group I/R); 1.1％ sevoflurane preconditioning group (group SP1); 2.2％ sevoflurane preconditioning group (group SP2); and 3.3％ sevoflurane preconditioning group (group SP3).Renal I/R was induced by clamping bilateral renal pedicles for 45 min followed by 6 h reperfusion.In SP1-SP3 groups,1.1％,2.2％ and 3.3％ sevoflurane were inhaled for 1 h,respectively,before ischemia,followed by 10 min washout.The rats were sacrificed by exsanguination at 6 h of reperfusion,and lungs were removed for determination of wet/dry lung weight (W/D) ratio,myeloperoxidase (MPO) activity and expression of angiotensin converting enzyme (ACE) mRNA,and for microscope examination of pathologic changes which were scored.Results Compared with group S,the pathological score,W/D ratio,MPO activity and expression of ACE mRNA were significantly increased in I/R and SP1-SP3 groups (P ＜ 0.05).Compared with group I/R,the pathological score,W/D ratio,MPO activity and expression of ACE mRNA were significantly decreased in SP1-SP3 groups (P ＜ 0.05).Compared with SP1 and SP3 groups,the pathological score,W/D ratio,MPO activity and expression of ACE mRNA were significantly decreased in group SP2 (P ＜ 0.05).Compared with group SP1,the W/D ratio was significantly decreased (P ＜0.05),and no significant changes were found in the other parameters in group SP3 (P ＞ 0.05).Conclusion Sevoflurane preconditioning can attenuate lung injury induced by renal I/R and inhibition of ACE synthesis is involved in the mechanism.
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Objective To investigate the expression of apoptosis-related genes Bcl-2 and Bax after cerebral ischemic reperfusion in hippocampal CA1 and CA3 areas and the effect of curcumin on the expression of Bcl-2 and Bax in gerbils.Methods Gerbils were randomly divided into sham group(SH),ischemia-reperfusion group(IR),curcumin group(CU) and solvent control group(SC).Forebrain ischemia was induced by occlusion of bilateral common carotid arteries.Observations were made in each group 1d,3d,5d and 7d after ischemia.Open field test was used to examine the behavioral change.The apoptotic neurons in hippocampal CA1 region was counted,and the expression of Bcl-2 and Bax in hippocampal CA1 and CA3 areas was detected by SABC immunocytochemical technique.Results The behavioral mark and the number of apoptotic neurons in hippocampal CA1 and CA3 areas were much smaller in CU group than in IR group(P
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Aim To explore the expression and role of ERK and JNK in cerebral ischemic preconditioning in gerbil.Methods Forebrain ischemia was induced by occlusion of bilateral carotid arteries.Gerbils were randomly divided into sham group(SH),ischemic-preconditioning control group(IC),ischemic-preconditioning group(IP),ischemia-reperfusion group(IR).The gerbils were killed in 15 min,2,4,6 h,1,3,5 and 7 d in each group following reperfusion.Open field test was used to examine the behavioral deficit in the due time.The number of surviving and apoptosis neurons in hippocampal CA1/3 region was counted,and the activity of p-ERK and p-JNK in hippocampus was detected using SP immunocytochemical technique.Results The behavioral mark and the number of apoptosis neurons in hippocampal CA1 region in group IP were much less than that in group IR(P