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.

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.

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.

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.

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.

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.

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.

Objective:To evaluate the effect of propofol on neuronal activity in medial prefrontal cortex (mPFC) during social behavior in sleep-deprived rats.Methods:Sixty SPF healthy male Sprague-Dawley rats, aged 8 weeks, weighing 200-250 g, were divided into 3 groups ( n=20 each) using a random number table method: control group (group Con), chronic sleep deprivation plus natural sleep group (group CSD+ NS), and chronic sleep deprivation plus propofol group (CSD+ Pro). Sleep deprivation model was developed by the modified multiple platform method, and the rats were placed in the sleep-deprivation tank for 20 h a day (14: 00-10: 00) and allowed to sleep naturally for 4 h (10: 00-14: 00) a day for 28 consecutive days.Propofol 40 mg/kg was intraperitoneally injected after sleep deprivation for 28 consecutive days in group CSD+ Pro, while the equal volume of 10% fat emulsion was given instead in group C and group CSD+ NS.Electroencephalographic recordings in cerebral cortical regions were performed on the days 1st, 14th and 28th after sleep deprivation.The apoptotic neurons in mPFC were detected using TUNEL method after the end of sleep deprivation, and the apoptosis index was calculated.A three chamber sociability test was used to detect the social behavior of rats, and local field potential signals in mPFC were collected. Results:Compared with group Con, the percentage of rapid eye movement sleep was significantly increased, the sniffing time preference coefficients in the 2 stages were reduced, the percentage of the β waves and θ waves-band power in mPFC during the social sniffing process was decreased, and the apoptosis index of neurons in mPFC was increased in group CSD+ NS ( P<0.05). Compared with group CSD+ NS, the percentage of rapid eye movement sleep was significantly increased, the sniffing time preference coefficient in the 2 stages was increased, and the percentage of β waves and θ waves-band power in mPFC during the social sniffing process was increased, and the apoptosis index of neurons in mPFC was decreased in group CSD+ Pro ( P<0.05). Conclusions:Propofol inhibits the apoptosis in neurons in mPFC and increases β and θ waves in the mPFC during social interaction after sleep deprivation in sleep-deprived rats, which is helpful in improving sleep deprivation-induced social disorder.

Objective:To evaluate the effect of propofol on parvalbumin (PV) neurons in the medical prefrontal cortex(mPFC)of rats with social behavior disorders induced by chronic sleep deprivation.Methods:Forty-two SPF male Sprague-Dawley rats, aged 8 weeks, weighing 200-250 g, were divided into 3 groups ( n=14 each) using a random number table method: control group (group Con), chronic sleep deprivation plus natural sleep group (group CSD+ NS), and chronic sleep deprivation plus propofol group (group CSD+ Pro). Sleep deprivation model was established by the modified multiple platform method, the rats were placed in the sleep-deprivation tank for 20 h a day (14: 00-10: 00), and allowed to sleep naturally for 4 h (10: 00-14: 00) a day for 28 consecutive days. Propofol 40 mg/kg was intraperitoneally injected for 28 consecutive days after sleep deprivation in CSD+ Pro group. While the equal volume of 10% fat emulsion was given in Con and CSD+ NS groups. After the end of sleep deprivation, a three-box social experiment was used to detect the social behavior of rats, and the number of the PV positive cells and density of the perineuronal network (PNN) in the mPFC area were measured by immunofluorescence. Results:Compared with group Con, the pertentage of rapid eye movement sleep and sniffing time preference coefficients for the strange rat 1 in the first stage and for the strange rat 2 in the second stage were significantly decreased, and the number of the PV positive cells and density of PNN in the mPFC area were decreased in group CSD+ NS ( P<0.05). Compared with group CSD+ NS, the sniffing time preference coefficients for the strange rat 1 in the first stage and for the strange rat 2 in the second stage were significantly increased, the number of the PV positive cells and density of PNN in the mPFC area were increased( P<0.05), and no significant change was found in the percentage of the rapid eye movement sleep in group CSD+ Pro. Conclusions:Propofol probably increases the number and function of PV neurons in the mPFC and ameliorates sleep deprivation-induced social behavior disorders in sleep-deprived rats.

Pituitary tumors are usually benign but can occasionally exhibit hormonal and proliferative behaviors. Dysregulation of the G1/S restriction point largely contributes to the over-proliferation of pituitary tumor cells. F-box protein S-phase kinase-interacting protein-2 (SKP2) reportedly targets and inhibits the expression of p27(Kip1), a well-known negative regulator of G1 cell cycle progression. In this study, SKP2 expression was found to be upregulated while p27(Kip1) expression was determined to be downregulated in rat and human pituitary tumor cells. Furthermore, SKP2 knockdown induced upregulation of p27(Kip1) and cell growth inhibition in rat and human pituitary tumor cells, while SKP2overexpression elicited opposite effects on p27(Kip1) expression and cell growth. The expression of microRNA-186 (miR-186) was reported to be reduced in pituitary tumors. Online tools predicted SKP2 to be a direct downstream target of miR-186, which was further confirmed by luciferase reporter gene assays. Moreover, miR-186 could modulate the cell proliferation and p27(Kip1)-mediated cell cycle alternation of rat and human pituitary tumor cells through SKP2. As further confirmation of these findings, miR-186 and p27(Kip1) expression were downregulated, while SKP2 expression was upregulated in human pituitary tumor tissue samples; thus, SKP2 expression negatively correlated with miR-186 and p27(Kip1) expression. In contrast, miR-186 expression positively associated with p27(Kip1) expression. Taken together, we discovered a novel mechanism by which miR-186/SKP2 axis modulates pituitary tumor cell proliferation through p27(Kip1)-mediated cell cycle alternation.
Animals ; Cell Cycle ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; Genes, Reporter ; Humans ; Luciferases ; Pituitary Neoplasms ; Rats ; Up-Regulation