Effects of heat shock protein 70 on inflammation after hepatic local ischemia/reperfusion in rats

AIM:To study the effects of heat shock protein 70 (HSP70) induced by the heat stress pretreatment on inflammation after hepatic ischemia/resperfusion.METHODS:With the hepatic local ischemia/reperfusion model (IR group),heat stress pretreatment (H+IR group) and injecting quercetin before heat stress pretreatment (Q+H+IR group) were performed.The HSP70,intercellular adhesion molecule-1 (ICAM-1) and the myeloperoxidase (MPO) activity were detected.The levels of serum ALT and AST and histological changes of the hepatocytes were also observed.RESULTS:In H+IR group,the HSP70 expression was higher than that in other groups at each time point,after performing ischemia-perfusion,hepatic injury was slighter.The levels of serum ALT and AST were increased slightly (P<0.01).The expression of ICAM-1 and the changes of MPO activity increased and peaked respectively at 6 h and 12 h after reperfusion.However,they were lower in H+IR group than those in IR group and Q+H+IR group.CONCLUSION:The HSP70 induced by heat stress pretreament reduces the expression of ICAM-1 and the changes of MPO activity during hepatic ischemia-reperfusion,then hepatic injury is depressed from the inflammation.     

Ischemic postconditioning inhibits activation of nuclear factor-κB and ICAM-1 expression in lungs following intestinal ischemia reperfusion in rats

AIM: To investigate the effect of ischemic postconditioning (I-postC) on the expression of nuclear factor-κB (NF-κB) and intercellular adhesion molecule-1 (ICAM-1) in the lungs following intestinal ischemia reperfusion(II/R) in rats, and to explore the possible mechanism of I-postC in attenuating lung injury induced by II/R. METHODS: Thirty-two male Wistar rats were randomly divided into sham, II/R, intestinal ischemic postconditioning (II-postC) and limb ischemic postconditioning (LI-postC) groups. The model of intestinal I/R injury was established by clamping the super mesenteric artery for 45 min followed by 120 min of reperfusion in rats. At the end of the experiment, the changes of arterial blood gas and lung index were measured, and the morphological changes of the lung tissues were observed under light microscope. The content of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and myeloperoxidase (MPO) in the lung tissues were also detected. The contents of NF-κB p65 and ICAM-1 in lung tissue were determined by immunohistochemical staining and Western blotting. RESULTS: (1) Compared to those in II/R group, II-postC and LI-postC improved the respiratory functions of the lung, characterized by the increase in PaO₂ and decrease in PaCO₂ (P<0.05 vs II/R group). The lung index was decreased (P<0.01) and the pathologic lesion of the lung tissues was alleviated significantly by II-postC and LI-postC. (2) Both II-postC and LI-postC markedly inhibited the decrease in SOD activity, the increase in the content of MDA and the activity of MPO in the lung tissues (P<0.05 or P<0.01) induced by intestinal I/R. In addition, the over-expression of NF-κB p65 and ICAM-1 in the lung tissues was inhibited markedly by II-postC and LI-postC (P<0.05 or P<0.01 vs II/R group). CONCLUSION: I-postC attenuates lung injury induced by intestinal I/R in rats due to suppressing the activation of NF-κB and subsequent accumulation of neutrophils mediated by ICAM-1.     

Inhibitory effect of NF-κB decoy ODN on expressions of TLR4 and IL-8 in LPS- induced intestinal epithelial cells

AIM: To study the effect of NF-κB decoy oligodeoxynucleotides(ODNs) on TLR4 and IL-8 expression in LPS-induced SW480 cells. METHODS: SW480 cells were cultured in vitro and stimulated for 3 h with LPS (10 μg/L). NF-κB decoy oligodeoxynucleotides mediated by lipofectin 2000 were added into the cell culture for 6 h. The supernatants were collected and messured for IL-8 by ELISA. TLR4 mRNA and IL-8 mRNA were examined by RT-PCR, respectively. The results were compared with control group, Scrambled ODNs group and lipofectin 2000 group. RESULTS: After SW480 cells were stimulated by LPS, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly increased, and the difference compared with control group was obvious. After treated with NF-κB decoy oligodeoxynucleotides, TLR4 mRNA, IL-8 mRNA and IL-8 expressions were significantly inhibited. The Scrambled ODNs group and lipofectin 2000 group had no effect on them. CONCLUSION: NF-κB decoy ODNs will become a new gene drug for treating inflammatory bowel disease(IBD).     

Effects of erigeron breviscapine on nuclear factor-κB expression following lung ischemia-reperfusion injury

AIM: To investigate the effects of erigeron breviscapine on nuclear factor-κB (NF-κB) expression following lung ischemia-reperfusion (I/R) injury in rats. METHODS: Thirty-two male Sprague-Dawley rats were randomized into four groups with 8 animals in each group: sham operation group (I), I/R group (II), erigeron 25 mg/kg group (III) and erigeron 50 mg/kg group (IV). A lung I/R injury rat model was established in situ. I/R injury consisted of 45 min of lung cross-clamping followed by 2 h of reperfusion; sham operation animals had a thoracotomy only. The wet-to-dry weight ratio (W /D), myeloperoxidase (MPO) of lung tissue, the content of nuclear NF-κB p65 were detected by immunohistochemical staining and Western blotting. The histopathological changes of lung tissue were observed under light microscopy. Electron microscopic evaluation was done on randomly selected lungs of two rats in each group at the end of the experiment. RESULTS: Compared to sham operation group, W /D and MPO in the I/R group increased significantly after reperfusion, and the expression of NF-κB in nucleus was up-regulated. As compared with I/R group, the level of NF-κB decreased in group III and IV. Also the changes of W /D and MPO were ameliorated as compared with group II. There was significant difference between group III and IV. CONCLUSION: Erigeron breviscapine reduced I/R lung injury through suppressing the activation of NF-κB and subsequent neutrophils accumulation.     

CO2 与养分交互作用对番茄幼苗养分利用的影响

以番茄(Lycopersicon esculentumMill.)‘合作906’为材料进行溶液培养试验,设2个因子:CO2和营养液浓度;CO2浓度设正常(360μL/L)和倍增(720μL/L)2个水平;营养液浓度设基本营养液(日本山崎番茄营养液),微量元素采用阿农营养液配方的1/2、1/4、1/8、1/164个水平,完全试验方案8个处理,3次重复。pH为6·0±0·2,3d更换1次营养液。移植到1·2L盆(2株/盒)中,植株在CO2生长箱(VS-3DMC)中培养,全天施放CO2,白天25℃,晚上15℃,光照为14h/d,光照强度11000lx,相对湿度60%。46d时收获,根、茎、叶经蒸馏水冲洗吸干水分后,放入纸袋105℃杀青,75…     

Exogenous carbon monoxide protects against the lung injury induced by ischemia-reperfusion of hind limbs in rats

AIM: To study the mechanism of protective effect of exogenous carbon monoxide (CO) in the lung injury induced by ischemia-reperfusion (IR) of hind limbs in rats. METHODS: Thirty-two SD rats were randomly divided into 4 groups: control, control+CO, IR and IR+CO. A rat model of ischemia in hind limbs and the reperfusion lung injury was made. The rats in IR+CO and control+CO groups were exposed to air containing 2.5×10-8 CO for 1 h before reperfusion or the corresponding control time point, while the other two groups were exposed to the routine air. The lung tissue structure, polymorphonuclear leukocyte (PMN) count, wet-to-dry weight ratio (W/D), malondialdehyde (MDA) content and the animal survival rate were observed. The carboxyhemoglobin (COHb) levels in artery blood were detected with CO-oximeter and the expression of intercellular adhesion molecule-1 (ICAM-1) in the lung was detected by Western blotting. RESULTS: Compared to control, the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly increased in IR group. Compared with the IR group, the blood COHb level was significantly increased and the animal mortality, lung PMNs number, W/D, MDA content and ICAM-1 expression were all significantly decreased in IR+CO group. CONCLUSION: These data suggest that exogenous CO attenuate limb IR-induced lung injury by down-regulatiny ICAM-1 expression and suppressing PMN sequestration in the lung following limb IR in rats.     

Effect of Ginkgo biloba extract on the expression of c-fos and HSP70 following focal cerebral ischemic reperfusion in rats

AIM: To investigate the influence of Ginkgo biloba extract (GBE) on the expression of c-fos, heat shock protein 70 (HSP70) during focal cerebral ischemic reperfusion in rats. METHODS: The middle cerebral artery occlusion (MCAO) model described by Zea longa was used. Healthy Wistar rats were randomized to 4 groups. Immunohistochemistry, in situ hybridization and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were used to detect the expression of c-fos gene, HSP70 and cell apoptosis at different reperfusion time points: 1, 6, 12, 24 hours and 3, 7 days after recirculation. RESULTS: The positive reactions of both c-fos and HSP70 were significantly increased at different reperfusion time in GBE-pretreated ischemia/reperfusion (IR) group than those in ischemia/reperfusion group (P<0.01) and the number of TUNEL-positive cells was reduced in GBE-pretreated IR group. CONCLUSION: The GBE induced the expression of c-fos, HSP70 and contributes to neuroprotective activities after cerebral ischemia.     

Ischemic postconditioning ameliorates pia mater microcirculation in rats subjected to cerebral ischemia reperfusion

AIM:To investigate the ameliorative effect of ischemic postconditioning (I-postC) on pia mater microcirculation in rats subjected to cerebral ischemia reperfusion (I/R) and its mechanisms.METHODS:Thirty-two male Wistar rats were randomly divided into sham, I/R, I-postC, and ischemic preconditioning (IPC) groups.The global cerebral I/R injury was induced by shunting carotid artery in rats.Pia mater microcirculation and cerebral microcirculatory perfusion were measured after reperfusion.The content of soluble intercellular adhesion molecule-1 (sICAM-1) in plasma was detected using enzyme linked-immunosorbent assay (ELISA).Myeloperoxidase (MPO), malondialdehyde (MDA), and superoxide dismutase (SOD) in cerebral tissue were detected.The expressions of vascular endothelial cell cadherin (VE-cadherin) and NF-κB p65 in cerebral tissue were assayed by Western blotting.RESULTS:(1) The disturbance of the blood flow in microvessel induced by I/R was improved significantly by I-postC.In addition, I-postC alleviated significantly the decrease in diameters of microvesseles, cerebral microcirculatory perfusion and cerebral VE-cadherin content induced by I/R (P＜0.05).(2) sICAM-1 in plasma, MPO and MDA in cerebral tissue decreased, but SOD activity in cerebral tissue increased in I-postC group, compared with those in I/R group (P＜0.05 or P＜0.01).The over-expression of NF-κB p65 induced by I/R was relieved by I-postC (P＜0.05).CONCLUSION:I-postC ameliorates pia mater microcirculation in rats subjected to cerebral I/R through suppressing the activation of polymorphonuclear neutrophils mediated by ICAM-1.     

Role of IRAK-4 activity in inhibitory effects of lipopolysaccharide pretreatment on hepatic ischemia/reperfusion injury

AIM:To observe the changes of interleukin-1 receptor associated kinase-4 (IRAK-4) in ischemia/reperfusion (I/R) liver pretreated with lipopolysaccharide (LPS) and to explore the protective mechanisms of LPS pretreatment against hepatic I/R injury. METHODS:Male Sprague-Dawley rats, weighing 240-280 g, were divided into three groups:control, ischemia/reperfusion group (I/R group) and LPS-pretreated group (LPS group). On the first day, LPS group received 0.1 mg/kg LPS via the tail vein, followed by 0.5 mg/kg on the 2nd, 3rd, 4th and 5th day. I/R group received the equivalent volumes (0.5 mL) of sterile PBS. Experiments of I/R injury was induced by temporary ischemia of the left lateral liver lobe for 90 min followed by 3 h reperfusion on 2 days after the last LPS treatment. At 0 min, 60 min and 180 min after reperfusion, the expression of IRAK-4 gene and protein level were determined by RT-PCR and Western blotting. The activity of NF-κB and the serum TNF-α level were also detected by ELISA. RESULTS:Although the level of IRAK-4 gene and protein were higher in the LPS group than that in I/R group and control group (P<0.01), no difference of the activities of NF-κB and the TNF-α level was observed between the LPS group and I/R group (P>0.05) at 0 min after reperfusion. However, all those indexes were evidently lower in the LPS group than those in I/R group (P<0.01) at 60 min and 180 min after reperfusion. CONCLUSION:This data suggests that the protective effects induced by LPS pretreatment against hepatic I/R injury may be via down-regulation of IRAK-4 expression.     

Hypothermic ventricular fibrillation with left ventricular drainage under cardiopulmonary bypass on canine lung preservation without aortic-cross clamping

AIM: To evaluate the protective effect of hypothermic ventricular fibrillation without aortic-cross clamping under cardiopulmonary bypass（CPB） on canine lung.METHODS: Fourteen dogs were randomly divided into two groups.All dogs received a standardized anesthetic technique.A conditional CPB was performed in every instance.Ventricular fibrillation was induced by systemic hypothermia to 28 ℃ and pericardial cooling saline in the experimental group.A standard CPB was performed in control group.The concentration of IL-8 in serum was measured by ELISA.The expressions of NF-κB and ICAM-1 were determined by using immunohistochemical staining.RESULTS: Serum IL-8 level in experimental group was significantly lower than that in control group (P<0.05).In experimental group,the pathological lesion of lung was improved.The results of histochemistry demonstrated that optical densities of NF-κB and ICAM-1 in two groups were much higher than those of pre-CPB (P<0.05),the optical densities of NF-κB and ICAM-1 in control group were much higher than those in experimental group (P<0.05).CONCLUSION: Hypothermic ventricular fibrillation under cardiopulmonary bypass may be an effective method on reducing SIRS and protecting pulmonary function.     

Effects of tirofiban on no-reflow and activation of NF-κB after myocardial ischemia/reperfusion in rats

AIM: To investigate the effects of platelet glycoprotein Ⅱb/Ⅲa receptor inhibitor tirofiban on myocardial no-reflow and activation of NF-κB after acute ischemia/reperfusion in rats. METHODS: Male Wistar rats were randomized into sham operation group, control group and tirofiban treatment group. Control group and tirofiban group were subjected ischemia for 90 min by ligation of coronary artery after thoracotomy and subsequently reperfusion for 120 min to establish acute myocardial ischemia/reperfusion no-reflow models. Thioflavine S, Evans blue and triphenyltetra zolium chloride (TTC) staining were performed to evaluate the area of no-reflow (ANR), infracted area (IA) and risk area (RA) of the heart. Immunohistochemistry was used for semi-quantitative analysis of the expression of nuclear factor-κB p65 (NF-κB p65) protein in myocytes and arteriole. Activity of myeloperoxidase (MPO) and content of malondialdehyde (MDA) in risk area of the heart were detected by ultraviolet spectrophotometer. RESULTS: After 120 min for reperfusion, compared to sham group, the statistical differences of higher positive expression of NF-κB p65 in myocytes and arteriole, activity of MPO and content of MDA both in control and tirofiban group were observed. Compared to control group, lower positive expression of NF-κB p65 in myocyte and arteriole, activity of MPO and content of MDA in tirofiban group were found (P<0.05, P<0.01). A markedly reduced ANR and IA were observed in tirofiban group than those in control group (34.36%±6.04% vs 52.09%±6.89%, P<0.01; 80.41%±8.48% vs 90.13%±5.72%, P<0.05). CONCLUSION: After myocardial ischemia/reperfusion for 120 min, no-reflow phenomenon can be observed in rats. Tirofiban reduces the areas of anatomic no-reflow and infarction, inhibits the activation of NF-κB in myocyte and arteriole, and decreases the infiltration of neutrophils and release of oxygen free radicals.     

Effects of safflor injection on expression of cyclooxygenase-2 mRNA during lung ischemia/reperfusion injury in rabbits

AIM: To investigate the effects of safflor injection (SI) on expression of cyclooxygenase-2 mRNA during lung ischemia/reperfusion injury (PIRI) in rabbits. METHODS: Rabbit lung model of ischemia/reperfusion injury was constructed in vivo. The rabbits were randomly divided into three groups: sham-operation group (group S), ischemia-reperfusion group (group I/R) and ischemia/reperfusion plus safflor injection group (group SI). The lung tissue sampled at the end of the experiment was assayed for wet/dry weight ratio (W/D), injured alveoli rate (IAR) and observed ultrastructure changes under electron microscope. The expressions of COX-1 and COX-2 were measured by immunohistochemistry (IHC). The expression of COX-1 mRNA and COX-2 mRNA were observed by in situ hybridization (ISH). RESULTS: The value of W/D and IAR was much higher in I/R group, but decreased in SI group. Electron microscope showed obvious ultrastructure injury brought by PIRI in I/R group, which was greatly attenuated in SI group. The IHC and ISH demonstrated that COX-2 and COX-2 expressions in pulmonary tissue of I/R group were significantly higher than those in S group (P<0.01). The difference of COX-1 and COX-1 expressions in pulmonary tissue was not observed among three groups. CONCLUSION: The lung ischemia-reperfusion insult induces the up-regulation of COX-2 in lung. Safflor injection may attenuate lung ischemia-reperfusion injury through inhibiting COX-2 expression.     

The expression of NF-κB and ICAM-1 in rat brain of experimental intracerebral hemorrhage and cerebral microvascular endothelial cells injured by hydrogen peroxide

AIM: To discuss the possible mechanism of the inflammation after intracerebral hemorrhage (ICH) and the relationship of nuclear factor-kappa B (NF-κB) and intercellular adhesion molecule-1 (ICAM-1). METHODS: The expression of NF-κB and ICAM-1 were detected by immunohistochemistry, in situ hybridization, immunocytochemistry and Western blotting techniques in rat brain of experimental ICH and cerebral microvascular endothelial cells (RCMECs) injured by hydrogen peroxide. RESULTS: The expression of NF-κB p65 and ICAM-1 were up-regulated in rat brain after ICH. The ICAM-1 reached the peak at 1 day while the NF-κB at 4th day. NF-κB p65 expressed remarkably in cultured RCMECs immediately after injured by hydrogen peroxide, while ICAM-1 expressed remarkably　2 hours later. PDTC, an inhibitor of NF-κB, down-regulated the expression of NF-κB and ICAM-1. CONCLUSION: NF-κB induces the expression of ICAM-1 in RCMECs injured by reactive oxygen species (ROS).     

Effect of limb ischemic postconditioning on brain focal ischemia and reperfusion injury in mice

AIM: To establish the mouse model in which the limbic ischemic postconditionning (LIPostC) enhances the tolerance against brain ischemia, and to investigate the effects of LIPostC on the ischemic extent and roles of heat shock protein 70 (HSP70) in ischemia and reperfusion injury. METHODS: The male Kunming mice were used in the study. The brain ischemia reperfusion (I/R) model was made by middle cerebral artery occlusion (MCAO). In the first test, the male mice were randomly divided into 9 groups (n=10): sham group, ischemia/reperfusion (I/R) groups (with ischemia for 0.5 h, 1 h,1.5 h and 2 h) and LIPostC+I/R groups (0.5 h+LIPostC,1 h+LIPostC,1.5 h+LIPostC,2 h+LIPostC). The reperfusion was performed after LIPostC for 24 h. After the neurologic deficit scores were evaluated, the brains were taken out to measure the infarct volume with TTC staining and to observe the pathological changes of cerebral cortex with HE staining. The neuronal apoptosis was determined by TUNEL. In the second test, the male mice were randomized into 4 groups (n=6): sham group, I/R group, LIPostC+I/R group and LIPostC+I/R+quercetin group (2 h ischemia). The neurological deficit scores were evaluated at 24 h after operation. The expression of HSP70 was determined by Western blotting.RESULTS: The duration of brain ischemia was related to the motor behavior and degree of brain injury. The longer the ischemic duration of the brain was performed, the more severe the pathological and behavioral changes were observed. The brain injury in 2 h MCAO mice was more severe than that in 1 h and 1.5 h MCAO mice (P<0.05). Compared to I/R group, each LIPostC group showed lower neurological score, less infarct volume and TUNEL positive neuron. The expression of HSP70 protein was increased and neurological functions were improved significantly in the mice with LIPostC. However, the neuroprotective role of LIPostC was attenuated by treating with quercetin, an inhibitor of HSP70.CONCLUSION: LIPostC promotes the expression of HSP 70, improves the neurological functions and attenuates the ischemia and reperfusion injury in MCAO mice. HSP70 produces a marked effect on the ischemic tolerance induced by LIPostC in MCAO mice.     

Effects of panax notoginseng saponins on pneumocyte apoptosis and Fas/FasL expression in rabbits with lung ischemia/reperfusion injury

AIM: AIM: To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury, and observe effects of panax notoginseng saponins (PNS) on apoptosis in lung ischemia/reperfusion injury. METHODS: Single lung in situ ischemia/reperfusion animal model was used. Eighty four Japanese white rabbits were randomly divided into control group (control), ischemia/reperfusion 1 h group (IR1h), IR3h, IR5h, Panax Notoginseng Saponins 1 h group (PNS1h), PNS3h and PNS5h. TUNEL, immunocytochemistry and in situ hybridization techniques were used to observe apoptosis and Fas/FasL expression in various phases of lung ischemia/reperfusion. RESULTS: Cell apoptosis in lung tissues were significantly high, Fas/FasL mRNA and its protein were up-regulated in lung tissues of lung ischemia/reperfusion injury compared with control (all of P<0.01). The PNS suppressed apoptosis as well as expression of Fas/FasL mRNA and its protein (P<0.05 or P<0.01, respectively). There was a significant correlation between expression of Fas/FasL protein, Fas/FasL mRNA and cell apoptosis (r=0.540，0.658，0.668，0.686；all P<0.01). CONCLUSIONS: Activation of Fas/FasL system and its initiating cell apoptosis of lung tissues may contribute to the pathogenesis of lung ischemia/reperfusion injury. The protective effects of PNS include suppressing the activation of Fas/FasL system and blocking apoptosis in lung tissues in lung ischemia/reperfusion injury.     

Irbesartan suppresses expression of VCAM-1 and ICAM-1 in myocardial ischemia-reperfusion areas in rabbits

AIM: To investigate the effects of irbesartan on acute myocardial ischemia-reperfusion injury and on expression of ICAM-1 and VCAM-1 in myocardial ischemia-reperfusion areas in rabbits. METHODS: Anterior descending branch of left coronary artery (LAD) was ligated in ischemia-reperfusion (IR) group but not in the sham group. Rabbits of IR group were subjected to 60 min of LAD occlusion, and 360 min of reperfusion. In the treated group, the rabbits were given irbesartan until the trial ended. Then, all rabbits were killed and tissue samples were removed from IR areas. After disposed with routine histological methods and stained with hematoxylin and eosin (HE), these samples were examined by light microscopy. The expression of ICAM-1 and VCAM-1 were also investigated by employing immunohistochemical SP method in the tissue samples. RESULTS: Degree of damage in cardiocytes and of neutrophil infiltration were more serious in IR group than those in sham group (P<0.01). Expression of ICAM-1 and VCAM-1 in IR areas of the treated group was much less up-regulation than that in IR group (P<0.01). CONCLUSION: The expression of ICAM-1 and VCAM-1 in I/R areas is up-regulated in the IR group. Irbesartan effectively alleviates myocardial IR injury in rabbits.     

Effects of Radix Angelicae Sinensis on changes of IL-6, TNF-α and bFGF in renal ischemia/reperfusion injury in rabbits

AIM: To determine the effect of Radix Angelicae Sinensis(RAS) on renal ischemia/reperfusion injury in rabbits and to explore its mechanism. METHODS: Twenty-five rabbits were divided randomly into the sham operated group(Control group), renal ischemia/reperfusion injury group(IR group) and RAS+IR group. At the time point of reperfusion 48 h after renal ischemia 1 h, the renal tissue were observed by electron-microscope and the contents of creatinine(Cr) in serum, tumor necrosis factor-α(TNF-α), interleukin-6(IL-6)and basic fibroblast growth factor(bFGF) in the renal tissue were measured. RESULTS: A remarkably degenerative changes in renal tissue were showed under electronmicroscope in IR group, but the changes in RAS+IR group were slight. The contents of Cr, TNF-α and IL-6 in IR group were higher than those in Control group, these parameters in RAS+IR group were lower than those in IR group, the difference between these groups was significant(P< 0.05 or P< 0.01). At the same time, the content of bFGF in IR group was lower than that in Control group(P< 0.01), while the content of bFGF in RAS+IR group was higher than that in IR group(P< 0.01) and Control group(P< 0.05). CONCLUSION: RAS has an effect of alleviating the renal ischemia/reperfusion injury by modulating the production or release of TNF-α, IL-6 and bFGF.     

Effects of human thioredoxin on pneumocyte apoptosis and Bcl-2/Bax expression in rats with lung ischemia/reperfusion injury

AIM: To explore the relationship between apoptosis in the lung tissues and lung ischemia/reperfusion injury, and to observe the effects of human thioredoxin (hTrx) on apoptosis in lung ischemia/reperfusion injury. METHODS: The single lung in situ ischemia/reperfusion animal model was used. Eighty four Wistar rats were randomly divided into control group (control), groups of ischemia for 1 h and reperfusion for different times (IR1h, IR3h, IR5h), and groups of IR+human thioredoxin treatment (IR1h +hTrx, IR3h +hTrx and IR5h +hTrx). Transmission electron microscope (TEM), terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and immunocytochemistry techniques were used to observe apoptosis, apoptosis signal-regulating kinase 1 (ASK1) and expression of Bcl-2 and Bax in various phases of lung ischemia/reperfusion. RESULTS: Cell apoptosis in lung tissues was significantly high, ASK1, Bcl-2 and Bax protein were up-regulated in lung tissues of lung ischemia/reperfusion injury as compared to control (all P<0.01). Compared to IR group, hTrx suppressed apoptosis as well as expression of ASK1 and Bax protein (P<0.01), Bcl-2 protein and the ratio of Bcl-2/Bax were up-regulated in lung tissues (all P<0.05 or P<0.01). There was a significant correlation between the expression of ASK1, Bax protein and cell apoptosis (r=0.775, r=0.814, respectively; all P<0.01). There was a negative correlation between cell apoptosis and Bcl-2/Bax protein (r=-0.275, P<0.05). CONCLUSION: Initiating cell apoptosis by the activation of Bcl-2/Bax system in lung tissues may contribute to the pathogenesis of lung ischemia/reperfusion injury. The protective effects of hTrx include suppressing the expression of ASK1, down-regulating the ratio of Bcl-2/Bax and blocking apoptosis in lung tissues in lung ischemia/ reperfusion injury.     

Pretreatment of hypertonic saline attenuates the hepatic ischemia reperfusion injury induced by neutrophils

AIM: To explore the effect of the pretreatment of hypertonic saline (HTS) in hepatic ischemia reperfusion (I/R) injury.METHODS: The rats were divided into sham group (sham group), ischemia reperfusion group (IR group) and pretreatment of hypertonic saline group (HTS group). Partial hepatic ischemia reperfusion model was used. The rats were sacrificed at the time of 1 h, 3 h, 6 h, 12 h and 24 h after reperfusion in each group, respectively. Blood samples were obtained to examine ALT. The expression of the CD11b/CD18 (Mac-1) on the neutrophils was analyzed by flow cytometry. RT-PCR and Western blotting were used to examine the expression of intercellular adhesion molecule-1 (ICAM-1) in livers and chromatometry was performed to detect the activity of myeloperoxidase (MPO) in livers. The morphology of hepatocytes and the structure of sinusoid were observed by histological examinations. RESULTS: ① HTS pretreatment decreased the level of ALT at the time points of 3 h, 6 h and 12 h after reperfusion (P<0.05). ② Mac-1 expression in HTS group was lower at 6 h and 12 h after reperfusion compared with IR group (P<0.05). ③ MPO activity in HTS group was lower at 6 h, 12 h and 24 h compared with IR group (P<0.05). ④ RT-PCR and Western blotting analysis indicated that the pretreatment of HTS inhibited the expression of ICAM-1 in livers after reperfusion. ⑤ Moderate hepatocyte swelling and few neutrophil infiltration were observed in HTS group.CONCLUSION: Pretreatment with HTS has the effect on hepatic ischemia reperfusion injury by inhibiting the expression of Mac-1 on circulating neutrophils and the expression of ICAM-1 in the liver.