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Objective To evaluate the role of mitochondrial fusion?fission in endotoxin?induced a?cute lung injury in rats. Methods Twenty healthy male Sprague?Dawley rats, weighing 160-180 g, were e?qually and randomly divided into either control group ( group C ) or endotoxin?induced acute lung injury group (group L) using a random number table. Lipopolysaccharide 5 mg∕kg was injected intravenously in group L, while the equal volume of normal saline 0?5 ml was given instead in group C. The animals were sacrificed at 6 h after administration of lipopolysaccharide or normal saline. The lungs were immediately re?moved for measurement of wet to dry lung weight ratio ( W∕D ratio) , superoxide dismutase ( SOD) activity and malondialdehyde ( MDA) content. The mitochondrial fusion proteins mitofusin 1 ( Mfn1) , Mfn2 and op?tic atrophy 1 ( OPA1) mRNA and protein expression was detected, and mitochondrial fission proteins dy?namin?related protein 1 (Drp1) and fission 1 (Fis1) mRNA and protein expression was also detected in lung tissues. Results Compared to group C, the W∕D ratio and MDA contents in lung tissues were signifi?cantly increased, SOD activity was decreased, Mfn1, Mfn2 and OPA1 mRNA and protein expression in lung tissues was down?regulated, and Drp1 and Fis1 mRNA and protein expression was up?regulated in group L. The pathological damage to lung tissues was obviously aggravated in group L when compared to group C. Conclusion The mechanism underlying endotoxin?induced acute lung injury is related to enhanced oxidative stress responses caused by decreased mitochondrial fusion and increased mitochondrial fission in rats.
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Objective To evaluate the relationship between erythroid 2-related factor (Nrf2 )-antioxidant response element (ARE) pathway and acute lung injury (ALI) induced by endotoxic shock in rabbits .Methods Thirty healthy male New Zealand white rabbits ,aged 2 months ,weighing 1.5-2.0 kg ,were randomly divided into 3 groups (n=10 each) using a random number table :control group (group C) ,group ALI and all-trans retinoic acid group (group ATRA ) .In group ATRA ,all-trans retinoic acid 6 mg/kg (in filter sterilized vegetable oil 1.2 ml) was injected intraperitoneally once a day for 2 days .ALI was induced by lipopolysaccharide 5 mg/kg (in normal saline 2 ml ) injected via the auricular vein at 10 h after the last injection of ATRA in ALI and ATRA groups .The equal volume of normal saline was injected instead in group C .The rabbits were sacrificed at 6 h after lipopolysaccharide or normal saline administration .The pulmonary specimens were removed for determination of wet/dry lung weight ratio (W/D ratio ) and expression of Nrf2 mRNA and nuclear protein ,and HO-1 mRNA and protein in lung tissues .The pathological changes of lungs were scored .Results Compared with group C ,the pathological score and W/D ratio were significantly increased , and the expression of Nrf2 mRNA and nuclear protein ,and HO-1 mRNA and protein was up-regulated in ALI and ATRA groups ( P 0.05 ) .Conclusion Activation of Nrf2-ARE pathway is the regulatory mechanism of the body adapting to the ALI induced by endotoxic shock in rabbits .
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Objective To evaluate the effect of electro-acupuncture (EA) at Zusanli and Feishu on endotoxin shock-induced acute lung injury in rabbits.Methods Sixty healthy male New Zealand white rabbits aged 2 months weighing 1.5-2.0 kg were randomly divided into 6 groups (n =10 each):group sham operation (group S); group zinc protoporphyrin-Ⅸ (ZnPP-Ⅸ) (group Z); group lipopolysaccharide (LPS) (group L); group LPS + EA (group EL) ; group LPS + sham EA (group SEL) and group LPS + EA + ZnPP-Ⅸ (group ELZ).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg and tracheostomized.The animals kept spontaneous breathing.Right internal carotid artery was cannulated for BP monitoring.Ear vein was cannulated for drug administration.LPS 5 mg/kg was injected iv in groups L,EL,SEL,ELZ.Endotoxin shock was confirmed by decrease in BP by 20 ％ of the baseline value and PaO2/FiO2 ≤ 300.ZnPP-Ⅸ (heme oxygenase (HO-1 ) inhibitor)10μmol/kg was injected intraperitoneal at 2 h after LPS injection in groups Z and ELZ.Bilateral 15 min EA stimulation of Zusanli and Feishu ( according to atlas of animal acu-points) was performed once a day for 5 days before LPS administration in groups EL and ELZ.The animals were sacrificed by blood-letting at 6 h after LPS administration.The lungs were removed for microscopic examination (0 =no injury,4 =most severe injury),detection of alveolar epithelial cell apoptosis (by TUNEL) and determination of HO-1 protein and mRNA expression.Results LPS significantly increased lung injury scores,alveolar epithelial cell apoptosis index (the number of apoptotic cells/total cells) and HO-1 protein and mRNA expression.EA significantly attenuated lung injury and alveolar epithelial cell apoptosis induced by LPS and further increased the expression of HO-1 protein and mRNA in group EL as compared with group L.The protective effects of EA was counteracted by ZnPP- Ⅸ in group ELZ.Conclusion EA at Zusanli and Feishu can attenuate endotoxin shock-induced lung injury by up-regulation of HO-1 expression and inhibiting alveolar epithelial cell apoptosis in the lung.
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ObjectiveTo evaluate the role of p38MAPK signaling pathway in the up-regulation of heme oxygenase-1 (HO-1) expression in the lung tissue in rats with acute lung injury (AL1) induced by endotoxic shock.MethodsForty-eight male SD rats,aged 8 weeks,weighing 180-200 g,were randomly divided into 4 groups ( n =12 each):control group ( group C ) ; endotoxic shock group ( group LS );endotoxic shock +SB203580 (a specific p38MAPK inhibitor) group (group LSS) and SB203580 group (group SB).Normal saline 0.5ml was injected via the femoral vein in groups C and SB,while LPS 10 mg/kg (in 0.5 ml normal saline) was injected via the femoral vein in groups LS and LSS.When MAP was decreased to 75％ of baseline value,10％ dimethyl sulfoxide (DMSO) 0.1 ml was infused via the femoral vein in groups C and LS,while SB203580 5 mol/kg (in 10％ DMSO 0.1 ml) was infused via the femoral vein at a rate of 0.01 ml/min in groups LSS and SB.Arterial blood samples were obtained at 6 h after LPS or normal saline was given for blood gas analysis and oxygenation index (PaO2/FiO2) was calculated.Then the rats were sacrificed and the lungs were removed for microscopic examination.The pathological changes of the lung were scored.The lung water content was calculated.The MDA content,SOD activity,and expression of HO-1 mRNA and protein,p38MAPK protein and phospharylated p38MAPK (p-p38MAPK) protein were determined.ResultsCompared with group C,the oxygenation index and SOD activity were significantly decreased,the pathological score,lung water content and MDA content were significantly increased,and the expression of HO-1 mRNA and protein and p-p38MAPK protein was significantly up-regulated ( P＜0.05),while no significant change was found in p38MAPK protein expression in groups LS and LSS,and no significant change was found in the indexes mentioned above in group SB (P＞0.05).Compared with group LS,the oxygenation index and SOD activity were significantly increased,the pathological score,lung water content and MDA content were significantly decreased,the expression of HO-1 mRNA and protein was significantly up-regulated,and p-p38MAPK protein expression was down-regulated ( P＜0.05),and no significant change was found in p38MAPK protein expression in group LSS ( P＞0.05).ConclusionThe inhibition of p38MAPK signaling pathway can lead to the up-regulation of HO-1 expression in lung tissues in rats with ALI induced by endotoxic shock.
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Objective To evaluate the role of activator protein-1 (AP-1) in the up-regulation of heme oxygenase-1 (HO-1) expression during lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats.Methods Forty-eight healthy male Sprague-Dawley rats,weighing 200-220 g,aged 2.5-3.0 months,were randomly divided into 4 groups (n=12 each): normal control group (group C),ALI group,curcumin + ALI group (group Cur+ ALI),and curcumin group (group Cur).In groups C and ALI,normal saline 0.5 ml and LPS 10 mg/kg (0.5 ml) were injected intravenously,respectively,30 min after 0.1％ dimethyl sulfoxide (the vehicle for curcumin) 0.5 ml was injected intraperitoneally.In groups Cur+ ALl and Cur,curcumin 20 mg/kg (0.5 ml) was injected intraperitoneally,and 30 min later LPS 10 mg/kg and normal saline 0.5 ml were injected,respectively.The rats were then sacrificed at 6 h after injection of LPS.The lungs were removed for microscopic examination.The pathological changes of the lung were scored.The malondialdehyde (MDA) content,superoxide dismutase (SOD)activity and expression of HO-1,AP-1 and HO-1 mRNA in lung tissues were determined.Results Compared with group C,the pathological score and MDA content were significantly increased,the SOD activity was significantly decreased,and the expression of HO-1,AP-1 and HO-1 mRNA was up-regulated in groups ALl and Cur +AL(l) (P ＜ 0.05),and no significant change was found in the parameters mentioned above in group Cur (P ＞ 0.05).The pathological score and MDA content were significantly higher,and the SOD activity and expression of HO-1,AP-1 and HO-1 mRNA were significantly lower in group Cur + ALl than in group ALI(P ＜ 0.05).Conclusion Transcription factor AP-1 activation is involved in the up-regulation of HO-1 expression during LPS-induced ALI in rats.
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Objective To evaluate the role of c-AMP-protein kinase A (cAMP-PKA) on the up-regulation of heme oxygenase-1 (HO-1) expression during lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats.Methods Forty-eight healthy male Sprague-Dawley rats,weighing 180-220 g,aged 2.5-3.0 months,were randomly divided into 4 groups (n =12 each)∶ normal control group (group C),ALI group (group ALI),H89 +ALI group (group H + ALI) and H89 group (group H).In group C,normal saline (solvent for LPS) 0.5 ml was injected via the femoral vein and normal saline (solvent for H89) 0.5 ml was injected subcutaneously 2 h later.In group ALI,10 mg/kg LPS 0.5 ml was injected via the femoral vein and normal saline 0.5 ml was injected subcutaneously 2 h later.In group H +ALI,10 mg/kg LPS 0.5 ml was injected via the femoral vein and 5 mg/kg H89 0.5ml was injected subcutaneously 2 h later.In group H,normal saline 0.5 ml was injected via the femoral vein and 5 mg/kg H89 0.5 ml was injected subcutaneously 2 h later.The rats were then sacrificed at 6 h after iv injection of LPS and the lungs were removed for microscopic examination and lung water content.The pathological changes of the lung were scored.The expression of HO-1 and PKA (by Western blot) and HO-1 mRNA (by RT-PCR) was detected.Results Compared with group C,the pathological score and lung water content were significantly increased,and the expression of HO-1,PKA and HO-1 mRNA was up-regulated in groups ALI and H +ALI (P ＜0.05),and no significant change was found in the parameters mentioned above in group H (P ＞ 0.05).The pathological score and lung water content were significantly higher,and the expression of HO-1,AP-1 and HO-1 mRNA was significantly lower in group H + ALI than in group ALI (P ＜ 0.05).Conclusion Activation of signaling pathway c-AMP-PKA is involved in the up-regulation of HO-1 expression during LPS-induced ALI in rats.
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Objective To investigate the effect of hydrogen gas on lipopolysaccharide (LPS)-induced apoptosis in human umbilical vein endothelial cells (HUVECs) in vitro.Methods HUVEC-12 cells were seeded in 96-well plates with a density of 1 × 104/ml (200 μl/hole) or in 6-well plates (2 ml/hole) with a density of 1 × 106/ml and randomly divided into 4 groups (n =30 each):control group (group C),hydrogen gas (H2) group,LPS group and LPS + H2 group.The cells were cultured in the plain culture medium in groups C and LPS or in hydrogen-saturated culture medium in groups H2 and LPS + H2.In addition,LPS 1 μg/ml was added simultaneously in groups LPS and LPS + H2 and the equal volume of normal saline was added instead in groups C and H2.The cell viability and apoptosis were measured by MTT assay and flow cytometry respectively after 24 h incubation.The concentration of high-mobility group box 1 (HMGB1) in the supernatant was determined by ELISA.Results Compared with groups C and H2,the cell viability was significantly decreased,and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly increased in groups LPS and LPS + H2 (P ＜ 0.05).Compared with group LPS,the cell viability was significantly increased,and the apoptotic rate and concentration of HMGB1 in the supernatant were significantly decreased in group LPS + H2 (P ＜ 0.05).Conclusion Hhydrogen gas can effectively reduce LPS-induced apoptosis in HUVECs through inhibiting the release of HMGB1.
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Objective:To study the influence of Taurine on proliferation of neural stem cells after focal cerebral ischemia in rats and to explore the mechanism of taurine in treating ischemic brain injury.Methods:The model of middle cerebral artery occlusion was made by using opening ranium hot coagulation in 80 rats,who were divided randomly into normal group,sham operate group,model group and taurine group.Taurine was administrated once daily by i.p.in taurine group,and immunohistochemistry method was used to observe the number of Brdu positive cells on the 3rd,7th,14th and 21st day after ischemia.Results:There were Brdu positive cells in the cortex,hippocampal dentate gyrus(DG) and striatum at different time points after cerebral ischemia,and the number of Brdu positive cells in various brain regions increased significantly on the 7th day as(18.65 ?4.38),(25.63?3.58),(19.46 ?2.98) and having statistical significance(P
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AIM: To investigate the effect of microwave ablation of liver cancer on the cellular immunity in mice. METHODS: A C57BL/6J mouse model of liver cancer was established by subcutaneous injection of Hepa 1 - 6 cells. The tumors were subjected to microwave ablation under the ablation condition of 45 ℃, 50 ℃, 55 ℃ or 60 ℃ for 180 s. The CD4~+ T cells, CD8~+ T cells and natural killer cells (NK) in peripheral blood were detected by FACS. The cytotoxicity of splenic NK and splenic cytotoxic T lymphocytes (CTL) activated by inactivated Hepa 1-6 cells was assayed by LDH method. RESULTS: The proportions of CD4~+ T cells, CD8~+ T cells and NK cells in peripheral blood in 50 ℃ and 55 ℃ group at 21 d after ablation were significantly increased and that of NK cells in 60 ℃ group was significantly increased. There was no significant difference between those in group 42 d after ablation and control. The cytotoxicities of splenic CTL and NK cells in 50 ℃ and 55 ℃ groups at 21 d or 42 d after ablation were significantly increased, and they were much higher than those in 45 ℃ group at the same time. The cytotoxicities of splenic CTL in 50 ℃ and 55 ℃ groups at 21 d after ablation were much higher than that in 60 ℃ group at the same time. CONCLUSION: Under a certain ablation temperature, microwave ablation of liver cancer promotes the cellular immunity.