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Objective:To evaluate the effect of artesunate on intestinal ischemia/reperfusion (I/R)-induced lung injury in mice and relationship with heme oxygenase-1 (HO-1).Methods:Twenty-four healthy SPF male C57BL/6 mice, aged 8-9 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (group Sham), intestinal I/R group (group I/R), artesunate group (group A), and artesunate plus HO-1 inhibitor Zinc protoporphyrin Ⅸ(ZnPP) group (group AS). The model of intestinal I/R injury was established by occluding the superior mesenteric artery for 45 min followed by 2 h reperfusion in anesthetized animals.Artesunate 40 mg/kg was injected via the tail vein at 1 h before ischemia in group A. ZnPP 7.5 mg/kg was injected via the tail vein at 12 h before ischemia, and artesunate 40 mg/kg was injected via the tail vein at 1 h before ischemia in group AS.The animals were sacrificed at the end of reperfusion, and the lung tissues were obtained for microscopic examination of the pathologic changes and for determination of the wet/dry lung weight (W/D) ratio, myeloperoxidase (MPO) activity, malondialdehyde (MDA) content, expression of interleukin-6 (IL-6) mRNA (by fluorescence quantitative polymerase chain reaction) and apoptotic index (AI) (by TUNEL). The lung injury score was assessed. Results:Compared with group Sham, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly increased, and the expression of IL-6 mRNA was up-regulated in group I/R ( P<0.05). Compared with group I/R, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly decreased, and the expression of IL-6 mRNA was down-regulated in group A ( P<0.05). Compared with group A, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly increased, and the expression of IL-6 mRNA was up-regulated in group AS ( P<0.05). Conclusions:Artesunate can alleviate intestinal I/R-induced lung injury, and the mechanism may be related to activation of HO-1 in mice.
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Objective:To evaluate the role of cannabinoid type 2 receptor (CB2R) in sevoflurane postconditioning-induced reduction of intestinal ischemia-reperfusion (I/R) injury in rats.Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 220-270 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (group Sham), intestinal I/R group (group I/R), intestinal I/R+ sevoflurane postconditioning group (group Sevo) and intestinal I/R+ sevoflurane postconditioning+ CB2R antagonist AM630 group (group AM). The model of intestinal I/R injury was established by occluding the superior mesenteric artery for 45 min followed by 2 h reperfusion.In the group Sevo, 2% sevoflurane was inhaled immediately at the beginning of reperfusion for 30 min.In the group AM, CB2R antagonist AM630 3 mg/kg was intraperitoneally injected at 1 h before ischemia, and the other treatments were similar to those previously described in group Sevo.At 2 h of reperfusion, the animals were sacrificed after anesthesia, and small intestinal tissues were obtained for microscopic examination of the pathologic changes which was scored according to Chiu and for determination of wet/dry weight ratio (W/D ratio), for detection of malondialdehyde (MDA) content (by thiobarbituric acid colorimetry), for determination of myeloperoxidase (MPO) activity (by MPO assay) and cleaved caspase-3 protein expression (by Western blot). Results:Compared with group Sham, the Chui score, W/D ratio, MDA content and MPO activity in the intestinal tissues were significantly increased, cleaved caspase-3 expression was up-regulated in group I/R ( P<0.05). Compared with group I/R, the Chui score, W/D ratio, MDA content and MPO activity in the intestinal tissues were significantly decreased, cleaved caspase-3 expression was down-regulated in group Sevo ( P<0.05). Compared with group Sevo, the Chui score, W/D ratio, MDA content and MPO activity were significantly increased, cleaved caspase-3 expression was up-regulated in group AM ( P<0.05). Conclusion:CB2R is involved in the process of sevoflurane postconditioning-induced reduction of intestinal I/R injury in rats.
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Objective To evaluate the role of phosphatidylinositol 3-kinase ( PI3K)∕protein kinase B ( Akt) signaling pathway in propofol-induced reduction of intestinal ischemia-reperfusion ( I∕R) injury in rats. Methods Thirty-two healthy male Sprague-Dawley rats, aged 2-3 months, weighing 225-275 g, were divided into 4 groups ( n=8 each) using a random number table method: sham operation group ( Sham group), intestinal I∕R group ( I∕R group), propofol group ( P group), and PI3K inhibitor wortmannin plus propofol group ( W+P group) . Intestinal ischemia was induced by occluding the superior mesenteric ar-tery for 45 min followed by 2 h of reperfusion to establish the model of intestinal I∕R injury. Propofol was in-travenously infused at a rate of 20 mg·kg-1 ·h-1 starting from the onset of reperfusion until the end of reper-fusion in group P. Wortmannin 15 μg∕kg was intravenously injected at 25 min before reperfusion, and propofol was intravenously infused at a rate of 20 mg·kg-1 ·h-1 starting from the onset of reperfusion until the end of reperfusion in group W+P. Rats were sacrificed at 2 h of reperfusion, and small intestinal tissues were obtained for microscopic examination of pathologic changes of intestinal mucosa and for determination of wet∕dry weight ratio (W∕D ratio), malondialdehyde (MDA) content (by thiobarbituric acid colorimetric method) , superoxide dismutase ( SOD ) activity ( using xanthine oxidase method ) , myeloperoxidase ( MPO) activity ( by MPO assay) , and phosphorylated Akt ( p-Akt) expression ( by Western blot) . Intes-tinal damage was assessed and scored according to Chiu. Results Compared with group Sham, Chiu' s score, W∕D ratio, MDA content and MPO activity were significantly increased, the SOD activity was de-creased, and p-Akt expression was down-regulated in group I∕R (P<0. 05). Compared with group I∕R, Chiu's score, W∕D ratio, MDA content and MPO activity were significantly decreased, the SOD activity was increased, and p-Akt expression was up-regulated in group P (P<0. 05). Compared with group P, Chiu's score, W∕D ratio, MDA content and MPO activity were significantly increased, the SOD activity was decreased, and p-Akt expression was down-regulated in group W+P (P<0. 05). Conclusion The mechanism by which propofol reduces intestrnal I∕R injury is related to activating PI3K∕Akt signaling path-way and inhibiting inflammatory and oxidative stress responses in rats.
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Objective To evaluate the effect of heme oxygenase-1 (HO-1) protein transduction on acute lung injury in septic rats.Methods Eighteen healthy male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly allocated into 3 groups (n =6 each) using a random number table:sham operation group (group Sham),sepsis group (group Sep),and fusion protein PEP-1-HO-1 group (group HO).Sepsis was produced by cecal ligation and puncture (CLP).In group HO,PEP-1-HO-1 fusion protein 0.6 mg was injected via the left iliac vein at 1 h before CLP and 5 h after CLP.The equal volume of normal saline was given instead of PEP-1-HO-1 in Sham and Sep groups.At 12 h after CLP,blood samples were collected from the right common carotid artery for measurement of serum tumor necrosis factoralpha (TNF-α) and interleukin-6 (IL-6) concentrations.The rats were then sacrificed,and lungs were removed for microscopic examination and for determination of wet/dry lung weight ratio (W/D ratio) and malondialdehyde (MDA) content (by thiobarbituric acid colorimetric method).Results Compared with group Sham,the W/D ratio and MDA content were significantly increased,the serum TNF-α and IL-6 concentrations were significantly increased (P＜0.05),and the pathological changes were significantly aggravated in Sep and HO groups.Compared with group Sep,the W/D ratio and MDA content were significantly decreased,the serum TNF-α and IL-6 concentrations were significantly decreased (P＜0.05),and the pathological changes were significantly attenuated in group HO.Conclusion HO-1 protein transduction can attenuate acute lung injury in septic rats,and the mechanism is probably related to inhibition of lipid peroxidation in lung tissues and systemic inflammatory responses.
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Objective To investigate the effects of heme oxygenase-1 (HO-1) protein transduction mediated by cell penetrating peptide PEP-1 on intestinal injury in a rat model of sepsis induced by cecal ligation and puncture (CLP).Methods Twenty-four healthy male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 4 groups (n =6 each) using a random number table:sham operation group (group S),group CLP,low-dose fusion protein PEP-1-HO-1 + CLP group (group P1) and high-dose fusion protein PEP-1-HO-1 + CLP group (group P2).Fusion protein PEP-1-HO-1 0.3 mg was administrated via the left iliac vein at 1 h before CLP and 5 h after CLP in group P1.Fusion protein PEP-1-HO-1 0.6 mg was administrated via the left iliac vein at 1 h before CLP and 5 h after CLP in group P2.The equal volume of normal saline was given instead of PEP-1-HO-1 in the other groups.The animals underwent laparotomy,but the caecum was not ligated or punctured in group S.Blood samples were collected at 12 h after CLP from the right common carotid artery for measurement of serum TNF-α and IL-6 levels.The rats were then sacrificed and intestines were removed for microscopic examination and for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in intestinal tissues.Results Compared with group S,the serum TNF-α and IL-6 levels,and MDA content in intestinal tissues were significantly increased,while SOD activity in intestinal tissues was decreased in CLP,P1 and P2 groups.Compared with group CLP,the serum TNF-α and IL-6 levels,and MDA content in intestinal were significantly decreased,while SOD activity in intestinal tissues was increased in P1 and P2 groups.Compared with group P1,the serum TNF-α and IL-6 levels,and MDA content in intestinal tissues were significantly decreased,while SOD activity in intestinal tissues was increased in group P2.The pathological changes of intestines were significantly mitigated in P1 and P2 groups as compared with group CLP.Conclusion HO-1 protein transduction attenuates intestinal injury induced by sepsis in rats,and the mechanism is related to inhibition of systemic inflammatory responses and lipid peroxidation in intestinal tissues.
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Objective To investigate the effects of heme oxygenase-1 (HO-1) transduced by cell penetrating peptide PEP-1 on renal ischemia/reperfusion (I/R) injury in rats.Methods Eighteen male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 3 groups (n =6 each):sham operation group (group S),renal I/R injury group (group I/R) and fusion protein PEP-1/HO-I + I/R group (group HO).I/R injury was produced by occluding bilateral renal arteries for 45 min followed by reperfusion for 6 h.The fusion protein PEP-1/HO-1 was injected via the left iliac vein 30 min prior to ischemia in group HO.Bilateral renal arteries were only exposed but not occluded in group C.Blood samples were collected from the right common carotid artery at 6 h of reperfusion for determination of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and HO-1 expression in renal tissues were measured.Results Compared with group S,the levels of MDA,serum BUN and Cr were significantly increased,the SOD activity was decreased,and HO-1 expression was up-regulated in groups I/R and HO (P ＜0.05).Compared with group I/R,the levels of MDA,serum BUN and Cr were significantly decreased,the SOD activity was increased,and HO-1 expression was up-regulated in group HO (P ＜ 0.05).Conclusion HO-1 protein can be successfully transduced into renal tissues by PEP-1 and transduced HO-1 protein reduces renal I/R injury by inhibiting lipid peroxidation response.
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Objective To evaluate the effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on liverinjury induced by intestinal ischemia/reperfusion (I/R) in rats.Methods Twenty-four male Sprague-Dawley rats,aged 7-9 weeks,weighing 210-260 g,were randomly divided into 3 groups (n =8 each):sham operation group (group S),intestinal I/R group (group I/R) and PEP-1/HO-1 group (group HO).To establish a model of intestinal I/R,intestines were exteriorized and the superior mesenteric artery was exposed and occluded for 45 min ischemia,and then the clamp was removed for 120 min reperfusion.The PEP-1/HO-1 fusion protein 0.5 mg was injectedvia ihe left iliac vein 30 min prior to ischemia in group HO.The superior mesenteric artery was only exposed but not occluded in group S.At the end of reperfusion,blood samples were collected from the right common carotid artery for measurement of serum aspartate aminotransferase (AST),alanine aminotransferase (ALT) activities.The rats were then sacrificed and livers were removed for microscopic examination and for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in livertissues.Results Compared with group S,serum AST and ALT activities and MDA content in liver tissues were significantly increased,while SOD activity in liver tissues was decreased in groups I/R and HO (P ＜ 0.05).Compared with group I/R,serum AST and ALT activities and MDA content in liver tissues were significantly decreased,while SOD activity in liver tissues was increased in group HO (P ＜0.05).Liver injury induced by intestinal I/R was significantly attenuated in group HO compared with group I/R (P ＜ 0.05).Conciusioon HO-1 protein mediated by cell penetrating peptide PEP-1 can attenuate liver injury induced by intestinalI/R in rats.
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Objective To investigate the effects of galantamine on the myocardial ischemia-reperfusion (I/R) injury in rats and the possible mechanism.Methods Fifty male SD rats weighing 225-275 g were randomly assigned into 5 groups (n =10 each):sham operation group (group SH); I/R group; galantamine + I/R group (group GAL); M receptor antagonist atropine + galantamine + I/R group (group AT); vagus nerve cut-off + galantamine + I/R group (group VGT).Myocardial I/R was induced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 120 min reperfusion.Normal saline 2 ml/kg was slowly injected via the femoral vein at 30 min before ischemia in groups SH and I/R.Galantamine 4 mg/kg was slowly injected via the femoral vein at 30 min before ischemia in group GAL.Atropine 4 mg/kg was slowly injected via the femoral vein at 45 min before ischemia in group AT and the other procedures were the same as those in group GAL.Bilateral cervical vagus nerves were cut off at 45 min before ischemia in group VGT and the other procedures were the same as those in group GAL.At the end of reperfusion,the hearts were removed for determination of myocardial infarct size,MPO and SOD activities,and MDA contents.Results The myocardial infarct size was significantly larger,the MPO activity and MDA content were significantly higher,and the SOD activity was significantly lower in group I/R than in group SH,and in groups AT and VGT than in group GAL (P ＜ 0.05).The myocardial infarct size was significantly smaller,the MDA content and MPO activity were significantly lower,and the SOD activity was significantly higher in group GAL than in group I/R P ＜ 0.05).Conclusion Galantamine has protective effect on myocardium against I/R injury and regulation of peripheral vagus nerve tension may be involved in the mechanism.
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Objective To investigate the effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on intestinal ischemia/reperfusion (I/R) injuiy in tats. Methods Eighteen male SD rats aged 7-9 weeks weighing 210-260 g were randomly divided into 3 groups (re = 6 each): sham operation group (group S) , I/R group and PEP-1/HO-1 + I/R group (group HO) . To establish a model of intestinal I/R injury, intestines were exteriorized and the superior mesenteric artery was exposed and occluded for 45 min ischemia, and then the clamp was removed for 120 min reperfusion. The PEP-1/HO-1 fusion protein 0.5 mg was injected via the left iliac vein 30 min prior to ischemia in group HO. The superior mesenteric artery was exposed but not occluded in group S. At the end of reperfusion, the rats were sacrificed and intestinal tissues obtained to determine the intestinal wet/ dry ratio, malondialdehyde (MDA) level, activities of superoxide dismutase (SOD) and HO-1, and HO-1 protein expression. The histological changes in the intestinal mucosa were examined and the injuiy was scored. Results Compared with group S, the intestinal wet/dry ratio, MDA level, HO-1 activity, HO-1 protein expression and injury score were significantly increased, while the SOD activity was significantly decreased in groups I/R and HO ( P ＜ 0.05) . Compared with group I/R, the intestinal wet/dry ratio, MDA level and injury score were significantly decreased, while the SOD activity, HO-1 activity and HO-1 protein expression increased in group HO ( P ＜ 0.05) . The pathologic changes were significantly attenuated in group HO compared with group I/R.Conclusion HO-1 protein can be successfully delivered into intestinal tissues by PEP-1 and has protective effects against intestinal I/R injury.
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Objective To investigate the effect of PEP-1-heme oxygenase-1 (PEP-1-HO-1) fusion protein transduction on hypoxia-reoxygenation (H/R) injury in rat H9c2 cells. Methods After construction of the prokaryotic expression plasmid pET15b-PEP-1-hHO-1 containing the human heme oxygenase-1 gene, it was then transformed to make PEP-1-HO-1 fusion protein express. The H9c2 cells were cultured in high-glucose Dulbecco's modified Eagle's medium (DMEM) supplemented with 15% fetal bovine serum and randomly divided into 4 groups (n = 4 each): control group (group C), H/R group, low-concentration fusion protein group (group L-HO), and high-concentration fusion protein group (group H-HO). The cells were exposed to 22 h of hypoxia followed by 8 h of reoxygenation. PEP-1-HO-1 fusion protein was added to the culture medium with a final concentration of 1.0 μ mol/L (group L-HO) or 2.0 μmol/L (group H-HO) before hypoxia. The cells and supernatant of the culture medium were collected after reoxygenation to determine the activity of lactate dehydrogenase (LDH) in the supernatant and the content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in the cells. Results The SOD activity was significantly lower, while the MDA content and LDH activity were significantly higher in group H/R, L-HO and H-HO than in group C (P ＜0.05). The SOD activity was significantly higher, while MDA content and LDH activity were significantly lower in group L-HO and H-HO than in group H/R, and in group H-HO than in group L-HO ( P ＜ 0.05). Conclusion PEP-1-HO-1 fusion protein transdution can protect H9c2 cells against H/R injury in rats.
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Objective To investigate the protective effects of heme oxygenase-1 (HO-1) mediated by cell penetrating peptide PEP-1 on myocardium against ischemia/reperfusion (IR) injury in isolated rat hearts. Methods Healthy male SD rats weighing 220-280 g were anesthetized with intraperitoneal pentobarbital. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5% CO2 at 37 ℃. Eighteen isolated rat hearts were randomly divided into 3 groups ( n = 6 each): Ⅰ group sham operation (group S);Ⅱ group IR and Ⅲ group PEP-1/HO-1 + IR (group HO-1). The isolated rat hearts were perfused with an oxygena-ted (95% O2-5% CO2 ) K-H solution at 37 ℃ in a Langendorff apparatus and were subjected to 40 min of global ischemia followed by 50 min of reperfusion after 30 min of stabilization. In group Ⅲ (group HO- 1 ) the isolated hearts were perfused with 50 μmol/L PEP-1/HO-1 for 15 min before ischemia. After 50 min of reperfusion, HO-1expression, MDA content and SOD activity in myocardial tissues were determined. The activities of creatine kinase (CK) and lactic dehydrogenase (LDH) in coronary effluent fluid were measured. Results The HO- 1 expression was significanfly higher in HO-1 group than in group IR. IR induced significant increase in MDA content and decrease in SOD activity in myocardium and CK and LDH activities in coronary effluent in group Ⅱ compared with group S. PEP-1/HO-1 significantly attenuated IR-induced changes. Conciusion HO-1 mediated by PEP-1 has protective effects on myocardium ngainst IR injury in rats.