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Purpose@#The role of vacuolar protein sorting 34 (Vps34), an indispensable protein required for cell vesicular trafficking, in the biological behavior of hepatocellular carcinoma (HCC) has yet to be studied. @*Materials and Methods@#In the present study, the expression of Vps34 in HCC and the effect of Vps34 on HCC cell invasion was detected both in vivo and in vitro. Furthermore, by modulating the RILP and Rab11, which regulate juxtanuclear lysosome aggregation and recycling endosome respectively, the underlying mechanism was investigated. @*Results@#Vps34 was significantly decreased in HCC and negatively correlated with the HCC invasiveness both in vivo and in vitro. Moreover, Vps34 could promote lysosomal juxtanuclear accumulation, reduce the invasive ability of HCC cells via the Rab7-RILP pathway. In addition, the deficiency of Vps34 in HCC cells affected the endosome-lysosome system, resulting in enhanced Rab11 mediated endocytic recycling of cell surface receptor and increased invasion of HCC cells. @*Conclusion@#Our study reveals that Vps34 acts as an invasion suppressor in HCC cells, and more importantly, the endosome-lysosome trafficking regulated by Vps34 has the potential to become a target pathway in HCC treatment.
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OBJECTIVES@#The integrated model of prenatal diagnosis and postnatal treatment for congenital heart disease (CHD) leads to an increasing number of operation in infants. This study aims to reveal the risk factors for postoperative early mortality and delayed recovery in infants less than 3 months old, who underwent surgical treatment for CHD in the Department of Cardiovascular Surgery, Second Xiangya Hospital, Central South University during the past 5 years.@*METHODS@#Clinical variables were collected via medical records. Delayed recovery was defined as the time of postoperative intubation, or cardiac intensive cure unit (CICU) stay, or hospital stay longer than its third quartile. Risk factors for early postoperative prognosis and the odds ratio (OR) were analyzed with logistic regression analysis.@*RESULTS@#A total of 511 infants underwent surgical treatment for CHD from January 2016 to June 2020 were retrospectively reviewed, including 217 (42.5%) infants with complex CHD. The median age was 60 days (3 hours-90 days); and median weight was 4.5 (1.7- 8.4 kg). There were 26 postoperative mortalities, making the incidence at 5.1%, including 5 (5/294, 0.7%) mortalities in patients with uncomplicated CHD, and 21 (9.6%) mortalities in patients with complex CHD. Based on multivariable analysis, risk factors for postoperative mortality were diagnosis of complex CHD (OR=5.53, P<0.001), weight under 4.0 kg (OR=9.86, P<0.001), preoperative symptoms (OR=3.17, P=0.012), and emergency operation (OR=11.66, P<0.001). The median time for postoperative intubation, CICU stay, and hospital stay were 21.0 (0.3-979.0) hours, 3.0 (0.5-91.0) days, and 11.5 (3.0-105.0) days, respectively. A total of 177 (34.6%) infants delayed recover, with risk factors including diagnosis of complex CHD (OR=3.41, P=0.001), weight under 4.0 kg (OR=4.55, P<0.001), and preoperative symptoms (OR=3.91, P<0.001).@*CONCLUSIONS@#Surgical treatment for infants (<3 months) with CHD is still a challenge, particularly for infants with complex CHD and weight under 4.0 kg. We can improve the prognosis of CHD treatment in infants by establishing the integrated model of prenatal diagnosis and postnatal treatment to choose the most suitable time window, avoid symptoms before surgery, and reduce emergency operation.
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Female , Humans , Infant , Middle Aged , Pregnancy , Heart Defects, Congenital/complications , Length of Stay , Postoperative Period , Prognosis , Retrospective Studies , Risk FactorsABSTRACT
Objective:To investigate the clinical effect of using free bilateral anterolateral thigh flaps(ALTF) in series to repair large area soft tissue defects of forearm.Methods:The clinical data of 11 patients with large soft tissue defects of forearm admitted in the Department of Plastic(Repair and Reconstruction) Surgery, Lishui Hospital of Zhejiang University from March 2014 to December 2021 were retrospectively analyzed, including 8 males and 3 females. Aged 36 to 68 years old, with an average of 48 years old. VSD treatment was performed after debridement, and until fresh removed 3 to 5 days after the operation. Until the wound was fresh. The wound was repaired with free bilateral ALTF in series until fresh. The size of the forearm wound was 18 cm×15 cm-28 cm×13 cm. The cut area of a single flap was 10 cm×8 cm-20 cm×13 cm. The series of bilateral flaps: One of the flaps was used as the proximal flap, and its vascular pedicle was anastomosed with the arteries and veins of the recipient area. The other flap was used as the distal flap, and the arteries and veins between the 2 ALTFs on both sides were anastomosed. The vascular pedicle beyond the distal flap was ligated or anastomosed to the distal end of the ulnar artery or the distal end of the radial artery. The flap and the surrounding skin of the recipient area were sutured immediately. The donor sites of the flap was closed directly. Periodic and regular outpatient follow-up was performed after operation and the clinical efficacy was analyzed.Results:All the flaps successfully survived after the surgery. The postoperative follow-up lasted for 6-18 months, 12 months in average. The flaps survived well with good soft texture, without swelling, the capillary reaction time was normal, without surface ulceration, in rosy colour and restored protective sensations. The hand function of the affected limb recovered well. The wound at donor sites healed well without complications. At the last follow-up, the hand function of the affected limb was evaluated by the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, the result was 7 in excellent and 4 in good. The patients were satisfied with the flap and the therapeutic effect.Conclusion:The free bilateral ALTF in series can be used to repair a wound surface with large area, and the donor site can be closed at the same time. It is an effective method to repair large soft tissue defect of forearm.
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Objective:To evaluate the effect of high-concentration hydrogen inhalation on sepsis-associated encephalopathy (SAE) in mice.Methods:Healthy male ICR mice, aged 6-8 weeks, weighing 20-25 g, were divided into 4 groups ( n=50 each) using the random number table method: sham operation group (Sham group), SAE group, sham operation plus high-concentration hydrogen group (Sham+ H 2 group), and SAE plus high-concentration hydrogen group (SAE+ H 2 group). SAE model was prepared by cecal ligation and puncture (CLP) in anesthetized animals.At 1 and 6 h after operation, Sham+ H 2 and SAE+ H 2 groups inhaled the mixture of hydrogen and oxygen (67% hydrogen-33% oxygen) for 1 h, and Sham and SAE groups inhaled the mixture of nitrogen and oxygen (67% nitrogen-33% oxygen) for 1 h. The postoperative 7-day survival rate was recorded.Cognitive function was assessed by Y maze test at days 3, 5 and 7 after operation.The mice were sacrificed at 24 h after operation, and hippocampal tissues were obtained for microscopic examination of the pathological changes of neurons in hippocampal CA1 region (with a light microscope) and for determination of normal neuron count, contents of tumor necrosis factor-ɑ (TNF-α) and high mobility group box-1 (HMGB1) (by enzyme-linked immunosorbent assay), mitochondrial membrane potential (MMP) (by fluorescence spectrophotometry) and content of mitochondrial ATP (by fluorescein-fluorescent enzyme luminescence method). Results:Compared with Sham group, the 7-day survival rate after operation, percentage of spontaneous alternation at each time point after operation, and the number of normal neurons were significantly decreased, the contents of TNF-ɑ and HMGB1 were increased, and the contents of ATP and MMP were decreased in SAE and SAE+ H 2 groups ( P<0.05), and no significant change was found in Sham+ H 2 group ( P>0.05). Compared with SAE group, the 7-day survival rate after operation, percentage of spontaneous alternation at each time point after operation, and the number of normal neurons were significantly increased, the contents of TNF-ɑ and HMGB1 were decreased, and the contents of ATP and MMP were increased in SAE+ H 2 group ( P<0.05). Conclusions:High-concentration hydrogen inhalation can reduce SAE, and the mechanism may be related to reduction of hippocampal inflammatory responses and improvement in mitochondrial function in mice.
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Objective@#To our knowledge, no definitive conclusion has been reached regarding the relationship between glucocorticoids and hypertension. Here, we aimed to explore the characteristics of glucocorticoids in participants with dysglycemia and hypertension, and to analyze their association with blood pressure indicators.@*Methods@#The participants of this study were from the Henan Rural Cohort study. A total of 1,688 patients 18-79 years of age were included in the matched case control study after application of the inclusion and exclusion criteria. Statistical methods were used to analyze the association between glucocorticoids and various indices of blood pressure, through approaches such as logistic regression analysis, trend tests, linear regression, and restricted cubic regression.@*Results@#The study population consisted of 552 patients with dysglycemia and hypertension (32.7%). The patients with co-morbidities had higher levels of serum cortisol ( @*Conclusions@#Serum deoxycortisol was positively correlated with systolic blood pressure, pulse pressure, mean arterial pressure, mean blood pressure, and mean proportional arterial pressure. Glucocorticoids (deoxycortisol and cortisol) increase the risk of hypertension in people with dysglycemia, particularly in those with T2DM.
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Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Blood Pressure , Case-Control Studies , China/epidemiology , Cohort Studies , Glucocorticoids/blood , Glycemic Load , Hydrocortisone/blood , Hypertension/etiology , Prevalence , Risk Factors , Rural PopulationABSTRACT
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 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 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 investigate the changes in FUNDC1/microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) signaling pathway during sepsis-induced liver injury in mice.Methods Tbirtytwo clean-grade healthy male C57BL/6 mice,aged 6 weeks,weighing 20-25 g,were divided into sham operation group (n =8) and sepsis group (n =24) using a random number table method.Sepsis was induced by cecal ligation and puncture.Blood samples were obtained at 24 h after operation in sham operation group and at 6,12 and 24 h after establishing the model in sepsis group for determination of concentrations of alanine aminotransferase and aspartate aminotransferase in serum.Mice were then sacrificed,and the right lobe of livers was removed for examination of the pathological changes and for determination of the expression of FUNDC1 and LC3 Ⅱ by Western blot.The mitochondria in the right lobe of livers were isolated to measure the respiratory function,and respiratory control rate was calculated.Results Compared with sham operation group,the concentrations of alanine aminotransferase and aspartate aminotransferase in serum and pathological scores were significantly increased,the respiratory control rate of mitochondria was decreased,the expression of FUNDC1 was down-regulated,and the expression of LC3 Ⅱ was up-regulated at each time point after establishing the model in sepsis group (P＜0.05).Conclusion The mechanism by which sepsis induces liver injury may be related to inhibiting activation of FUNDC1/LC3 Ⅱ signaling pathway in mice.
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Objective To evaluate the effect of hydrogen on blood brain barrier of mice with sepsisassociated encephalopathy (SAE).Methods A total of 100 adult male ICR mice,aged 6-8 weeks,weighing 20-25 g,were divided into 4 groups (n =25 each) using a random number table method:sham operation group (group Sham),sham operation plus hydrogen group (group Sham+H),group SAE and SAE plus hydrogen group (group SAE+ H).Sepsis was induced by cecal ligation and puncture (CLP).Sham+H and SAE+H groups inhaled 2％ hydrogen for 1 h starting from 1 and 6 h after CLP,respectively.At 24 h after CLP,Evans blue (EB) was injected via the caudal vein,and then the mice were sacrificed and brain tissues were removed for measuring the EB and water contents,for examining the pathological changes of hippocampi (with a light microscope) and for detecting the expression of occludin and VE-cadherin (by Western blot).Morris water maze test was performed at days 10-16 after CLP.Results Compared with group Sham,the contents of EB and water in brain tissues were significantly increased,the expression of occludin and VE-cadherin was down-regulated,the escape latency was prolonged,and the number of crossing the original platform was reduced in SAE and SAE+H groups (P＜0.05).Compared with group SAE,the contents of EB and water in brain tissues were significantly decreased,the expression of occludin and VE-cadherin was up-regulated,the escape latency was shortened,the number of crossing the original platform was increased (P＜0.05),and the pathological changes of hippocampi were significantly attenuated in group SAE+H.Conclusion The mechanism by which hydrogen mitigates SAE may be related to reducing the damage to blood brain barrier of mice.
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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 relationship between heme oxygenase-1 (HO-1) expression and autophagy during acute lung injury in septic mice.Methods Forty-eight male C57BL/6 mice,aged 6-8 weeks,weighing 20-25 g,were randomly divided into 4 groups (n=12 each) using a random number table:sham operation group (group SH),sepsis group (group CLP),HO-1 agonist hemin plus sepsis group (group Hemin+CLP) and HO-1 inhibitor SnPP plus sepsis group (group SnPP+CLP).Sepsis was induced by cecal ligation and puncture in CLP,Hemin+CLP and SnPP+CLP groups.Hemin 50 μmol/kg and SnPP 50 μmol/kg were intraperitoneally injected at 2 h before operation in Hemin + CLP group and SnPP+CLP group,respectively.Arterial blood samples were taken at 24 h after operation for blood gas analysis,PaO2 was recorded,and oxygenation index was calculated.Lungs were removed for examination of pathological changes which were scored and for determination of autophagosome count (with an electron microscope),weight/dry weight ratio (W/D ratio) and expression of HO-1,Becling-1 and microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) in lung tissues (by Western blot).Results Compared with group SH,oxygenation index was significantly decreased,and W/D ratio,autophagosome count and pathological scores were increased in CLP,Hemin+CLP and SnPP+CLP groups,and the expression of HO-1,Beclin-1 and LC3 Ⅱ was significantly up-regulated in CLP and Hemin+CLP groups (P＜0.05).Compared with group CLP,oxygenation index and autophagosome count were significantly increased,W/D ratio and pathological scores were decreased,and the expression of HO-1,Beclin-1 and LC3 Ⅱ was up-regulated in group Hemin+CLP,and oxygenation index and autophagosome count were significantly decreased,W/D ratio and pathological scores were increased,and the expression of HO-1,Beclin-1 and LC3 Ⅱ was downregulated in group SnPP+CLP (P＜0.05).Conclusion Up-regulated expression of HO-1 mediates autophagy,which is involved in the endogenous protective mechanism underlying acute lung injury in septic mice.
ABSTRACT
Lead exposure is a known potential risk factor for neurodegenerative diseases such as Alzheimer's disease (AD). Exposure to lead during the critical phase of brain development has been linked with mental retardation and hypophrenia in later life. This study was aimed to investigate the effects of lead exposure of pregnant mice on the expressions of insulin-degrading enzyme (IDE) and nerve growth factor (NGF) in the hippocampus of their offspring. Blood samples were collected from the tail vein, and after anesthetizing the pups, the brain was excised on postnatal day 21. Lead concentrations were determined by graphite furnace atomic absorption spectrophotometry, and the expressions of IDE and NGF were determined by immunohistochemistry and Western blotting. Results showed that the reduction in IDE and NGF expression in the hippocampus of pups might be associated with impairment of learning and memory and dementia induced by maternal lead exposure during pregnancy and lactation.
Subject(s)
Animals , Female , Mice , Pregnancy , Down-Regulation , Gene Expression Regulation, Developmental , Gene Expression Regulation, Enzymologic , Hippocampus , Metabolism , Insulysin , Genetics , Metabolism , Lead , Toxicity , Prenatal Exposure Delayed EffectsABSTRACT
OBJECTIVES@#To study the differential genes expression in the early stage of acute renal ischemia-reperfusion injury and explore potential molecular mechanisms.@*METHODS@#The ischemia-reperfusion model was made via clamping renal artery of rat. The microarray detection and bioinformatics analyzing of the genes expression were performed. Differentially expressed genes were screened and related cellular activities and signaling pathways were analyzed in early stage of acute kidney injury. Meanwhile, molecules closely relative to acute kidney injury were explored by establishing a biological network of the differentially expressed genes, and the results were verified by real-time PCR.@*RESULTS@#A total of 151 genes showed differential expression in this study, including 132 up-regulated and 19 down-regulated genes. Cell proliferation, cytokines mediated signaling transduction and immune responses were greatly enriched by GO and KEGG analysis. The results of real-time PCR showed that compared with control groups, three selected genes （ANXA1, PHLDA1 and KLF6） which related to the acute kidney injury had an obvious differential expression in the early stage of disease. The multiple of increase was essentially the same as the multiple detected by microarray.@*CONCLUSIONS@#This study shows differential gene expression profile, related biological processes and signaling pathways involved in the early stage of acute kidney injury. ANXA1, PHLDA1 and KLF6 may play a role in the pathogenesis of acute kidney injury.