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Objective:To evaluate the relationship between the second messenger cyclic GMP-AMP (cGAS)-cyclic GMP-AMP receptor stimulator of interferon genes (STING) signaling pathway and ferritinophagy in the early stage of cerebral ischemia-reperfusion (I/R) in mice.Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 21-25 g, were divided into 4 groups ( n=6 each) using a random number table method: sham group, cerebral I/R injury group (CIRI group), cerebral I/R injury + cGAS inhibitor group (CIRI + RU group), and cerebral I/R injury + cGAS inhibitor + overexpressed nuclear receptor coactivator 4 (NCOA4) group (MCAO + RU + LV-NCOA4 group). The model of cerebral I/R injury was developed using the middle cerebral artery occlusion (MCAO) in anesthetized animals.In CIRI+ RU group, cGAS inhibitor 5 mg/kg was intraperitoneally injected at 10 min before reperfusion.In CIRI+ RU+ LV-NCOA4 group, NCOA4-overexpressing lentivirus (1×10 9 TU/ml) 2 μl was injected into the ventricle at 7 days before MCAO, and the other operations were the same as those previously described in CIRI+ RU group.After 6 h of reperfusion, the neurological function deficits were assessed and scored, then the mice were sacrificed, and brains were removed for determination of the cerebral infarct size (by TTC method), MDA content (by TBA method), activity of SOD (by WST-1 method), and expression of cGAS, STING, NCOA4, ferritin, and microtubule-associated protein 1 light chain 3B (LC3B) (by Western blot). Results:Compared with Sham group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05). Compared with CIRI group, the neurological function deficit score and cerebral infarct size were significantly decreased, SOD activity was increased, MDA content was decreased, the expression of cGAS, STING, NCOA4 and LC3B was down-regulated, and the expression of ferritin was up-regulated in CIRI+ RU group ( P<0.05). Compared with CIRI+ RU group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05), and no significant change was found in the expression of cGAS and STING in CIRI+ RU+ LV-NCOA4 group ( P>0.05). Conclusions:The cGAS-STING signaling pathway can promote the over-activation of ferritinophagy, enhance oxidative stress, and thus induce early CIRI in mice.
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Objective:To evaluate the role of B/glycogen synthase kinase-3β (Akt/GSK-3β) signaling pathway in interleukin-4 (IL-4)-induced reduction of cerebral ischemia-reperfusion (I/R) injury in mice and the relationship with autophagy.Methods:Forty clean-grade healthy male Balb/c mice, aged 10-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), cerebral I/R group (group IR), IR plus IL-4 group, and IR plus IL-4 plus Akt inhibitor LY294002 group (IR+ IL-4+ LY group). Cerebral I/R was induced by 60 min middle cerebral artery occlusion followed by 24 reperfusion in anesthetized mice.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model in group IL-4.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model, and LY294002 15 nmol/kg was simultaneously injected via the tail vein in group IR+ IL-4+ LY.Neurological function was assessed and scored at 24 h of reperfusion, and then animals were sacrificed and brains removed for determination of cerebral infarct size (by TTC assay), cell apoptosis, autophagosome count (with transmission electron microscope), levels of superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) (using colorimetric assay), phosphorylation of Akt and GSK-3β, expression of LC3 and Beclin-1 (by Western blot). The apoptosis index and LC3Ⅱ/LC3Ⅰ ratio were calculated. Results:Compared with Sham group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in the other three groups ( P<0.05). Compared with IR group, the neurological scores, cerebral infarct size and apoptosis index were significantly decreased, the SOD activity in brain tissues was increased, levels of MDA and ROS were decreased, phosphorylation of Akt and GSK-3β was increased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were decreased in IR+ IL-4 group ( P<0.05), and no significant change was found in the parameters mentioned above in IR+ IL-4+ LY group ( P>0.05). Compared with IR+ IL-4 group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in IR+ IL-4+ LY group ( P<0.05). Conclusion:IL-4 can inhibit cell autophagy through activating Akt/GSK-3β signaling pathway and thus reduces cerebral I/R injury in mice.
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Objective To evaluate the role of protein kinase B∕glycogen synthase kinase-3 beta (Akt∕GSK-3β) signaling pathway in trichostatin-A (TSA)-induced reduction of cerebral ischemia-reperfu-sion ( I∕R) injury in mice. Methods Forty pathogen-free healthy male Balb∕c mice, weighing 18-22 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group ( S group), I∕R group, TSA group and TSA plus Akt inhibitor LY294002 group (TL group). Cerebral I∕R was induced by middle cerebral artery occlusion ( 1-h ischemia followed by 24-h reperfusion) . TSA 5 mg∕kg was intraperitoneally injected for 3 consecutive days before establishing the model in TSA group. TSA 5 mg∕kg was intraperitoneally injected for 3 consecutive days before establishing the model, and LY29400215 nmol∕kg was injected via the caudal vein at 30 min before establishing the model. Brain tissues were ob-tained at 24 h of reperfusion for determination of cerebral infarct size ( by TTC ) , activities of superoxidedismutase ( SOD) and reactive oxygen species ( ROS) and malondialdehyde ( MDA) content ( by colorimet-ric assay), cell apoptosis (by TUNEL) and expression of Akt, phosphorylated Akt (p-Akt), GSK-3βand phosphorylated GSK-3β ( p-GSK-3β) . The apoptosis index and ratios of p-Akt∕Akt and p-GSK-3β∕GSK-3β were calculated. Results Compared with S group, the cerebral infarct size was significantly in-creased, the activity of SOD in brain tissues was decreased, the MDA content and ROS activity in brain tissues and apoptosis index were increased, and the ratios of p-Akt∕Akt and p-GSK-3β∕GSK-3β were de-creased in I∕R group ( P<0. 05) . Compared with I∕R group, the cerebral infarct size was significantly de-creased, the activity of SOD in brain tissues was increased, the MDA content and ROS activity in brain tis-sues and apoptosis index were decreased, and the ratios of p-Akt∕Akt and p-GSK-3β∕GSK-3β were de-creased in TSA group ( P<0. 05) . Compared with TSA group, the cerebral infarct size was significantly in-creased, the activity of SOD in brain tissues was decreased, the MDA content and ROS activity in brain tissues and apoptosis index were increased, and the ratios of p-Akt∕Akt and p-GSK-3β∕GSK-3β were de-creased in TL group ( P<0. 05) . Conclusion The mechanism by which TSA attenuates cerebral I∕R injury is related to activating Akt∕GSK-3β signaling pathway in mice.
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Objective To investigate the effect of dexmedetomidine hydrochloride ( Dex ) preconditioning on renal function in rats after renal ischemia reperfusion injury under high glucose condition. Methods SD rats were randomly divided into 6 groups:NG-Sham operated group,NG-I/R group, NG-Dex group,HG-Sham operated group,HG-I/R group,HG-Dex group. Renal ischemia reperfusion model was established except Sham groups. Dex 50 μg·kg-1 was injected intraperitoneally 30 min before ischemia in the Dex preconditioning group,25% glucose 3 g·kg-1 was given intraperitoneally before the renal ischemia reperfusion model was established in high glucose groups. Blood glucose and renal function of each group were detected . Renal pathologic changes were observed with hematoxylin-eosin staining. Apoptosis of renal tissue was detected by TUNEL method. Results BUN,Cr and apoptosis rate in NG-I/R group were higher than those in NG-Sham operated group ( P<0.05);BUN,Cr and apoptosis rate in NG-Dex group were lower than those in NG-I/R group ( P<0.05);BUN,Cr and apoptosis rate in HG-I/R group and HG-Dex group were higher than those in NG-I/R group and NG-Dex group,respectively (P<0.05); However,there was no significant difference between HG-I/R group and HG-Dex group ( P>0.05) . Conclusion Dex has a protective effect on renal function after renal ischemia reperfusion, but this effect is inhibited in high glucose condition, which may relate to the increasing of kidney cells apoptosis.
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Objective To compare the ameliorating effect of collagen peptide chelated calcium (CPCC) and estrogen on the bone quality in ovariectomized rats in order to serve the development of safe drugs for prevention of osteoporosis (OP).Methods Bilateral ovariectomized rats were divided into ovariectomized group (OVX),sham group,17β-estradiol injection group (OVX+E2) and CPCC gavage group (OVX+CCCP).Bone and serum indices of these groups were assessed and compared at 9 weeks after treatment.Results Bone density of the OVX group was significantly lower than the sham group (P0.05),while the body weight gain of the E2 group at weeks 8 and 9 was significantly lower than those of the sham group (P<0.01).As regarding the prevention of bone loss,the Mg and Ca levels of the E2 group were significantly lower than those of the moderate and high dose CPCC groups.The Cu level was not significantly different compared with the sham group,while those in the moderate and low dose CPCC groups were significantly higher than the sham group.The Mn,Zn and hydroxyproline levels of the E2 group were significantly lower than those of the sham group,while the CPCC group maintained levels similar to that of the sham group.In regarding to the inhibiting effect on the increased blood BGP and StrACP,the E2 group was still maintained at levels similar to that of the OVX group,while those of the CPCC group were significantly lower than the OVX group.As regarding the decreased blood Ca,the E2 group was not significantly different with that of the OVX group,while that of the CPCC group was significantly higher than the OVX group.Conclusions CPCC is more effective than estrogen in ameliorating the bone quality of ovariectomized rats.
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Objective:To investigate the effect of different ventilation time in the prone position on patients with endogenous/exogenous ARDS.Methods:30 endogenous/30 exogenous ARDS patients were randomly devided into 4 groups,ventilation in the prone position for 2 h and 4 h.Recording the score of APCHEII,oxygenation index,the absorption situation in X-ray,HR,MAP,extubation time,the time out of ICU.Results:The APCHEII scores HR and MAP in four groups have no significant statistics (P＞0.05);4h ventilation for endogenous ARDS patients has a better indicators than 2 h in oxygenation index,the absorption situation in X-ray,extubation time and the time out of ICU (P＜0.05);2 h and 4 h ventilation for exogenous ARDS patients can improve indicators above,two groups have no significant statistics (P＞0.05),the results of exogenous groups are precede than endogenous group (P＜0.05).Conclusion:Ventilation in the prone position can improve the situation of ARDS patients,both endogenous patients and exogenous patients.Exogenous ARDS patients have a better treatment effect after the ventilation of 2h,however,endogenous patients need longer time and have a non-ideal prognosis.
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Objective To investigate the effect of dexmedetomidine hydrochloride ( Dex ) preconditioning on renal function in rats after renal ischemia reperfusion injury under high glucose condition. Methods SD rats were randomly divided into 6 groups:NG-Sham operated group,NG-I/R group, NG-Dex group,HG-Sham operated group,HG-I/R group,HG-Dex group. Renal ischemia reperfusion model was established except Sham groups. Dex 50 μg·kg-1 was injected intraperitoneally 30 min before ischemia in the Dex preconditioning group,25% glucose 3 g·kg-1 was given intraperitoneally before the renal ischemia reperfusion model was established in high glucose groups. Blood glucose and renal function of each group were detected . Renal pathologic changes were observed with hematoxylin-eosin staining. Apoptosis of renal tissue was detected by TUNEL method. Results BUN,Cr and apoptosis rate in NG-I/R group were higher than those in NG-Sham operated group ( P<0.05);BUN,Cr and apoptosis rate in NG-Dex group were lower than those in NG-I/R group ( P<0.05);BUN,Cr and apoptosis rate in HG-I/R group and HG-Dex group were higher than those in NG-I/R group and NG-Dex group,respectively (P<0.05); However,there was no significant difference between HG-I/R group and HG-Dex group ( P>0.05) . Conclusion Dex has a protective effect on renal function after renal ischemia reperfusion, but this effect is inhibited in high glucose condition, which may relate to the increasing of kidney cells apoptosis.
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Objective To investigate the effects of penehyclidine hydrochloride on activities of nuclear factor kappa B (NF-kB) and activator protein-1 (AP-1) during actue lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty male Sprague-Dawley rats,aged 8 weeks,weighing 250-300 g,were randomly assigned into 3 equal groups (n =10 each) using a random number table:sham operation group (group S),blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloride group (group PHCD).The model of actue lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until MAP was decreased to 35-45 mmHg within 15 min and maintained at this level for 60 min,followed by resuscitation.In PHCD group,PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established,blood samples were obtained for measurement of concentrations of tumor necrosis factor-alpha (TNF-α) in serum.The lungs were then removed for determination of lung water content,myeloperoxidase (MPO) activaty (by colorimetric assay),NF-κB and AP-1 activaties (using electrophoretic mobility shift assay) in lung tissues,and for microscopic examination of pathologic changes (under light microscope).The left lung was lavaged,and lung permeability index (LPI) was calculated.Results Compared with S group,lung water content,LPI,serum TNF-α level and activites of MPO,NF-κB and AP-1 were significantly increased in THSR and PHCD groups.Compared with THSR group,lung water content,LPI,serum TNF-α concentrations and activites of MPO,NF-κB and AP-1 were significantly decreased in PHCD group.The pathological damage to lung tissues was significantly reduced in PHCD group as compared with THSR group.Conclusion PHCD can inhibit activities of NF-κB and AP-1 in lung tissues,thus mitigating acute lung injury induced by blunt chest trauma-HSR in rats.
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Objective To investigate the effects of penehyclidine hydrochloride on Fas/FasL expression during acute lung injury induced by blunt chest trauma-hemorrhagic shock and resuscitation (HSR) in rats.Methods Thirty male SPF Sprague-Dawley rats, aged 8 weeks, weighing 245-275 g, were randomly assigned into 3 equal groups using a random number table: sham operation group (group Sham) , blunt chest trauma-HSR group (group THSR) and penehyclidine hydrochloric group (group PHCD).The model of acute lung injury induced by blunt chest trauma-HSR was induced by dropping a 300 g weight onto a precordium in anesthetized rats.Blood was withdrawn via the femoral artery 5 min later until mean arterial pressure was decreased to 35-45 mmHg within 15 min, and maintained at this level for 60 min, followed by resuscitation.In PHCD group, PHCD 2 mg/kg was injected intravenously at 60 min after hemorrhagic shock.At 6 h after the model was established, the rats were sacrificed, the lungs were then removed for microscopic examination of pathologic changes and for determination of Fas, FasL and caspase-8 expression, and interleukin-6 (IL-6) and IL-1β contents in lung tissues.Apoptotic index was calculated.Results Compared with group Sham, the expression of Fas, FasL and caspase-8 was significantly up-regulated, and AI and contents of IL-6 and IL-1β were increased in THSR and PHCD groups (P＜O.05).Compared with group THSR, the expression of Fas, FasL and caspase-8 was significantly down-regulated,and AI and contents of IL-6 and IL-1β were decreased in group PHCD (P＜0.05).The pathologic changes of lungs were significantly reduced in group PHCD compared with group THSR.Conclusion The mechanism by which penehyclidine hydrochloride inhibits lung cell apoptosis induced by blunt chest trauma-HSR is associated with inhibition of Fas/FasL expression in rats.
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Objective To investigate the mechanism of myocardial ischemia and reperfusion-induced acute lung injury (ALl) and protective effect of ischemic postconditioning.Methods Forty SD rats were allocated to sham group,myocardial ischemia/reperfusion group (reperfusion group),ischemic postconditioning group (postconditioning group),and ischemic postconditioning + phosphatase and tensin homolog deleted on chromosome ten (PTEN) inhibiting group (inhibitor group) according to the random number table,with 10 rats per group.Myocardial ischemia/reperfusion was induced by left anterior descending coronary artery occlusion.Postconditioning was performed within 1 minute before reperfusion consisting of 3 10 s cycles of reperfusion followed by 10 s occlusion.Lung was immediately removed 120 minutes after reperfusion for HE stain,immunohistochemical detection of inflammatory factors and apoptosis factors,TUNEL assay of cell apoptosis,and Western blot of protein kinase B (Akt),phospho-Akt (p-Akt),glycogen synthase kinase-3β (GSK-3β),and phospho-GSK-3β (p-GSK-3β).Results Down-regulated B-cell lymphoma-2 (Bcl-2) and IL-10 and up-regulated Bcl-2 associated X protein (Bax),cysteinyl aspartate specific proteinase-3 (Caspase-3),IL-6 as well as IL-8 were observed in other 3 groups compared with sham group (P ＜0.01).Moreover,down-regulated Bax,Caspase-3,IL-6,IL-8 as well as TUNEL and up-regulated Bcl-2 as well as IL-10 were observed in reperfusion group compared to postconditioning group and tensor group (P ＜ 0.01).No statistical differences were found among the four groups in levels of Akt,p-Akt,and GSK-3β,but level of p-GSK-3β was significantly down-regulated in reperfusion group compared to other 3 groups(P ＜ 0.01).Conclusion Development of ALI may relate to down-regulation of p-GSK-3β evoked directly by the release of inflammation factors in early period of myocardial ischemia/reperfusion and ischemic postconditioning may attenuate the condition.
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Objective To evaluate the effects of ischemic postconditioning on brain injury induced by myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Diabetes mellitus was induced by intraperitoneal streptozotocin 60 mg/kg and confirmed by blood glucose level ＞ 16.7 mmol/L.Thirty male Sprague-Dawley rats,weighing 220-280 g,in which diabetes mellitus was successfully induced,were randomly allocated into 3 groups (n =10 each) using a random number table:group sham operation (group S),group I/R and group ischemic postconditioning (group P).Myocardial I/R was induced by occlusion of the anterior descending branch of the left coronary artery in I/R and P groups.Group P received 3 cycles of 10 s reperfusion followed by 10 s ischemia at the end of myocardial ischemia.The rats were sacrificed at 120 min of reperfusion and the brains were removed for microscopic examination and for determination of cell apoptosis (by TUNEL) and expression of interleukin-6 (IL-6),IL-8,IL-10,glycogen synthase kinase-3 beta (GSK-3β) and phosphorylated GSK-3β (pGSK-3β) (by immuno-histochemistry).Apoptotic index was calculated.Results Compared with group S,apoptotic index was significantly increased,IL-6 and IL-8 expression was up-regulated,and IL-10 and pGSK-3β expression was downregulated in I/R and P groups (P ＜ 0.01).Compared with group I/R,apoptotic index was significantly decreased,IL-6 and IL-8 expression was down-regulated,and IL-10 and pGSK-3β expression was up-regulated in group P (P＜0.01).There was no significant difference in GSK-3β expression among the 3 groups (P ＞ 0.05).The pathologic changes were significantly attenuated in group P as compared with group I/R.Conclusion Ischemic postconditioning can attenuate brain injury induced by myocardial I/R in diabetic rats,and inhition of activity of GSK-3β may be involved in the mechanism.
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Objective To systematically review the efficacy of bispectral index (BIS) monitoring for prevention of intraoperative awareness in patients under general anesthesia.Methods The Cochrane Central Register of Controlled Trials (Central),PubMed,Medline,and EMBASE were searched for randomized controlled clinical trials involving detection of intraoperative awareness in patients in whom BIS was used or not under general anesthesia.The quality of the studies was evaluated by the method recommended by Cochrane Collaboration.Evaluation indexes included the incidence of intraoperative awareness.Meta-analysis was conducted using RevMan 5.1 software.Results Five randomized controlled clinical trials involving 34181 patients were included in this meta-analysis.There were 17432 cases in whom BIS was applied and the incidence of intraoperative awareness was 0.132％.There were 16749 cases in whom BIS was not used and the incidence of intraoperative awareness was 0.245％.There was no significant difference in the incidence of intraoperative awareness between the two groups (P ＞0.05).Further analysis was performed according to the method of anesthesia.In inhalational anesthesia,there were 13288 cases in whom BIS was applied and the incidence of intraoperative awareness was 0.128％,and there were 13202 cases in whom BIS was not applied and the incidence of intraoperative awareness was 0.113％.There was no significant difference in the incidence of intraoperative awareness between the two groups (P ＞ 0.05).In total intravenous anesthesia,there were 4144 cases in whom BIS was applied and the incidence of intraoperative awareness was 0.145％,and there were 3547 cases in whom BIS was not applied and the incidence of intraoperative awareness was 0.733 ％.The incidence of intraoperative awareness was significantly lower in the patients in whom BIS was applied than those in whom BIS was not applied (P ＜ 0.01).Conclusion BIS monitoring can effectively prevent the development of intraoperative awareness in patients under total intravenous anesthesia,but can not prevent the development of intraoperative awareness in patients under inhalational anesthesia.
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Objective To investigate the effects of ischemic pre- and postconditioning on cerebral glycogen synthase kinase-3 beta (GSK-3β) activity in a rat model of global cerebral ischemia-reperfusion (I/R).Methods Forty male Wistar rats weighing 200-230 g were randomly allocated into 4 groups (n =10 each) : Ⅰ group sham operation (group S); Ⅱ group I/R; Ⅲ group ischemic preconditioning (group IPR) and Ⅳ group ischemic postconditioning (group IPO). The animals were anesthetized with intraperitoneal 10% chloral hydrate 0.4 ml/100 g. Global cerebral ischemia was induced by four-vessel-occlusion in group Ⅱ , Ⅲ and Ⅳ. Bilateral vertebral arteries were cauterized and bilateral carotid arteries were occluded for 10 min. In group IPR cerebral ischemia was preceded by 3 cycles of 10 s ischemia followed by 30 s reperfusion. The group IPO received 3 cycles of 30 s reperfusion followed by 10 s ischemia at the end of 10 min cerebral ischemia. The animals were killed 2 days later. The brains were immediately removed for determination of neuronal apoptosis in the cortex (by TUNEL), the infarct size (by TTC), p-GSK-3β activity (by spectrum assay) and the expression of Bcl-2, Bax and Caspase-3 (by SP). Linear correlation of p-GSK-3β activity with the number of apoptotic neurons in the cortex and cerebral infarct size was analyzed. Results Cerebral I/R significantly increased the number of apoptotic neurons in the cortex and infarct size, decreased p-GSK-3β activity, down-regulated Bcl-2 expression and up-regulated Bax and Caspase-3 expression in group I/R as compared with group S. Ischemic pre- and postconditioning significantly attenuated these cerebral I/R-induced changes. The p-GSK-3β activity was negatively correlated with the number of apoptotic neurons in the cortex and cerebral infarct size. Conclusion Ischemic pre- and postconditioning reduces cerebral I/R injury through inhibiting the activity of GSK-3β.