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Sufficient embryos are needed for the preservation of Beagle dogs germplasm resources and the prepara-tion of gene?modified human disease animal models. Up to now, the induced ovulation technique has no effect on dogs,it is hard to obtain mature oocytes in vivo, although the scientists try a lot in many aspects, but still could not make a break?through. The in vitro maturation rate is too low to support the preservation of germplasm resources, application in gene?modified disease models and biomedical research. Aiming to provide useful information on breakthrough in dog oocytes mat?uration, this review will summarize the effect of different age and reproductive stage,different morphology and size of the oo?cytes and lipid droplet on the in vitro maturation of dog oocytes.
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Objective To study the effect of electroacupuncture pretreatment on cognitive function in aged rats and its possible mechanism. Methods 60 healthy male SD rats were randomly divided into 3 groups(n =20):control group (group C),simple operation group (S group),operation + electroacupuncture stimulation group (E Group),E group in the preoperative electroacupuncture stimulation Baihui、Neiguan、Qu pool and Zusanli,1 d,continuous stimulation 7 d. Group S and E were treated with hepatic ischemia-reperfusion. The water maze test (MWN)was performed at 1d before operation and 1,3,5 d and 7 d after operation,and the escape latency and swimming distance were recorded. The expression of Wnt protein ,β-catenin and phosphorylated glycogen synthase kinase-3β(p-GSK-3β) in cerebrospinal fluid were measured. Results Compared with group C ,the escape latency and swimming distance were significantly prolonged in group S and E at 1-7d(P<0.05),and the contents of Aβand p-tau-181 in S and E group were increased(P<0.05),the contents of Wnt protein,β-catenin protein and p-GSK-3β protein in group E were significantly increased (P < 0.05),and the contents of Wnt protein ,β-catenin and p-GSK-3βprotein in group S were significantly decreased(P<0.05);Compared with group S,the escape latency and swimming distance were significantly shortened in group E (P < 0.05),the levels of Aβ and p-tau-181 protein in group E were decreased(P<0.05),and Wnt protein andβ-catenin protein in group E were significantly lower than those in group E (P < 0.05),and p-GSK-3β protein were significantly increased (P <0.05). Conclusion Pretreatment with electroacupuncture can significantly improve the cognitive function of aged rats,the mechanism may be related to the activation of Wnt/β-catenin signaling pathway.
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Objective To study the effect of electroacupuncture pretreatment on cognitive function in aged rats and its possible mechanism. Methods 60 healthy male SD rats were randomly divided into 3 groups(n =20):control group (group C),simple operation group (S group),operation + electroacupuncture stimulation group (E Group),E group in the preoperative electroacupuncture stimulation Baihui、Neiguan、Qu pool and Zusanli,1 d,continuous stimulation 7 d. Group S and E were treated with hepatic ischemia-reperfusion. The water maze test (MWN)was performed at 1d before operation and 1,3,5 d and 7 d after operation,and the escape latency and swimming distance were recorded. The expression of Wnt protein ,β-catenin and phosphorylated glycogen synthase kinase-3β(p-GSK-3β) in cerebrospinal fluid were measured. Results Compared with group C ,the escape latency and swimming distance were significantly prolonged in group S and E at 1-7d(P<0.05),and the contents of Aβand p-tau-181 in S and E group were increased(P<0.05),the contents of Wnt protein,β-catenin protein and p-GSK-3β protein in group E were significantly increased (P < 0.05),and the contents of Wnt protein ,β-catenin and p-GSK-3βprotein in group S were significantly decreased(P<0.05);Compared with group S,the escape latency and swimming distance were significantly shortened in group E (P < 0.05),the levels of Aβ and p-tau-181 protein in group E were decreased(P<0.05),and Wnt protein andβ-catenin protein in group E were significantly lower than those in group E (P < 0.05),and p-GSK-3β protein were significantly increased (P <0.05). Conclusion Pretreatment with electroacupuncture can significantly improve the cognitive function of aged rats,the mechanism may be related to the activation of Wnt/β-catenin signaling pathway.
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Objective To evaluate the effects of age factors on hypothermia-induced reduction of ischemia-reperfusion(I/R)injury in isolated rat hearts.Methods Pathogen-free healthy aged male Sprague-Dawley rats,aged 18-20 months,weighing 400-600 g,and young rats,aged 4-6 months,weighing 280-350 g,were used in the study.After the animals were anesthetized with intraperitoneal chloral hydrate and heparinized,their hearts were excised and perfused with K-H solution in a Langendorff apparatus.Twenty-four isolated hearts of aged rats were assigned into 2 groups(n=12 each)using a random number table:I/R group(group AI/R)and hypothermia group(group AH).Twenty-four isolated hearts of young rats were assigned into 2 groups(n=12 each)using a random number table:I/R group(group YI/R)and hypothermia group(group YH).Perfusion was suspended for 30 min followed by 120 min reperfusion to establish the model of I/R.The temperature was maintained at 37 ℃ during the whole process in AI/R and YI/R groups.The hearts were perfused with 34 ℃ K-H solution until 120 min of reperfusion starting from onset of reperfusion in AH and YH groups.At 30 min of equilibration(T0)and 15,30,60 and 120 min of reperfusion(T1-4),heart rate(HR),left ventricular developed pressure(LVDP),the maximum rate of increase in left ventricular pressure(+dp/dtmax),and the minimum rate of increase in left ventricular pressure(+dp/dtmin)were recorded.Six hearts from each group were randomly selected at T4,and myocardial specimens were obtained for determination of ATP,superoxide dismutase(SOD)and malondialdehyde(MDA)levels and myocardial infarct size(IS).Results Compared with group YI/R,HR was significantly decreased at T1-4,ATP and SOD levels were increased,and the MDA content and myocardial IS were decreased in group YH,and the HR,LVDP,+dp/dtmax and +dp/dtmin at T0 and ATP and SOD levels at T4 were significantly decreased,and the MDA content and myocardial IS were increased in group AI/R(P<0.05).Compared with group YH,HR,LVDP,+dp/dtmax and +dp/dtmin at T0 and ATP and SOD levels at T4 were significantly decreased,and the MDA content and myocardial IS were increased in group AH(P<0.05).Compared with group AH,the levels of ATP and SOD were significantly decreased,and the MDA content and myocardial IS were increased in group AI/R(P<0.05).Conclusion Age factors affect the efficacy of hypothermia in reducing I/R injury in isolated rat hearts,and hypothermia provides better cardioprotection for young rats than for aged rats.
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Objective To investigate the effects of different duration hypotension thresholds on p-Tau-181 and Aβ-42 protein expression and cognition in rats. Methods Thirty-nine healthy male SD rats were randomly di-vided into 4 groups:the control group(group C,n=9),the hypotension group(groupA1、A2、A3 ,n=10). The blood pressure of groupA1、A2、A3 was measured in different time of 2 h、4 h、6 h ,for 5 days. The antihyperten-sive group of mean arterial pressure(MAP)were maintained in the 50~55 mmHg safe range. Morris water maze was used to detect the spatial learning and memory ability of rats. The levels of Aβ42 and p-Tau-181 were detected by ELISA. Results There was no significant difference in mortality of rats in each group (P > 0.05). Compared with the group C,the escape latency and swimming distance of A2 group and A3 group were increased(P<0.05). In 3~7 days after operation,the cerebrospinal fluid P-Tau-181 and Aβ42 protein expression increased in the A2 group and A3 group compared with the A1 group(P<0.05). The escape latency and swimming distance of the A2 group and the A3 group were significantly longer than those in the control group. Aβ42 and p-Tau-181 were signifi-cantly increased in A3 group(P < 0.05). Compared with the A2 group,the increase of Aβ42 and p-Tau-181 in the A3 group was not significant(P<0.05). Conclusion Long-term controlled hypotension may lead to postoper-ative cognitive dysfunction which may relate to the increase of Aβ42 and p-Tau-181 protein expression.
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Objective To evaluate the role of Wnt/β-catenin signaling pathway in lithium chloride preconditioning-induced improvement in postoperative cognitive function of aged rats.Methods A total of 100 pathogen-free healthy male Sprague-Dawley rats,aged 18 months,weighing 540-650 g,were divided into 5 groups (n =20 each) using a random number table:control group (group C),surgery group (group S),lithium chloride preconditioning group (group L),secreted frizzled-related protein 1 (sFRP-1) group (group F) and lithium chloride preconditioning plus sFRP-1 group (group L+F).Lithium chloride 2 mmol/kg was intraperitoneally injected once a day for 5 consecutive days before operation in L and L+ F groups.In F and L + F groups,sFRP-1 10 μl (concentration 10 μg/ml) was injected into the ventricle at 1 day before operation.Ten rats in each group were randomly sacrificed at 1 day after operation,and the hippocampi were removed for determination of the expression of phosphorylated glycogen synthase kinase-3 beta (p-GSK-3β),3-catenin and Wnt in hippocampal tissues (by Western blot).The rest rats underwent Morris water maze test at day 3-7 after operation,and the concentrations of amyloid beta 42 (A3-42) and phosphorylated tau-181 protein (p-tau-181) in cerebrospinal fluid (CSF) were detected by enzyme-linked immunosorbent assay at day 7 after operation.Results Compared with group C,the escape latency was significantly prolonged at 3-7 days after operation,and the concentration of Aβ-42 in CSF was increased in the other four groups,and the expression of Wnt,p-GSK-3β and β-catenin in hippocampal tissues was down-regulated in group S,the expression of Wnt,p-GSK-3β and β-catenin in hippocampal tissues was significantly up-regulated in L and L+F groups,and the concentration of p-tau-181 in CSF was significantly increased in S and L+F groups (P＜0.05).Compared with group S,the escape latency was significantly shortened at 3-7 days after operation in group L and at 5-7 days after operation in group L+F,and the expression of Wnt,p-GSK-3β and β-catenin was significantly up-regulated,and the concentrations of Aβ-42 and p-tau-181 in CSF were decreased in L and L+F groups (P＜0.05).Compared with group L,the escape latency was significantly prolonged at 3-6 days after operation,the expression of Wnt,p-GSK-3β and β-catenin was down-regulated,and the concentration of p-tau-181 in CSF was increased in group L+F (P＜ 0.05).Compared with group F,the escape latency was significantly shortened at 3-7 days after operation,the expression of Wnt,p-GSK-3β and β-catenin was up-regulated,and the concentrations of Aβ-42 and ptau-181 in CSF were decreased in group L+F (P＜0.05).Conclusion The mechanism by which lithium chloride preconditioning improves in postoperative cognitive function is partially related to activating Wnt/β-catenin signaling pathway in aged rats.
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Objective To investigate the effects of Saikosaponin A (SSA)on cognitive function and cAMP/CREB signaling pathway and expression of BDNF in mice after traumatic brain injury. Methods Sixty SD male mice were randomized into three groups:shame operation group (group S, n =20),trauma group (group T,n =20)and SSA treatment group (group A,n =20).Mice received an administration of SSA 5 mg/kg (group A)or equal volume saline (group S,group T)immediately and once daily for 5 consecutive days after trauma.The cognitive function was detected by Morris wa-ter maze test on day 1,3,7 and 14 after trauma.The hippocampal tissues were harvested after be-havioral tests and homogenized for measuring the levels of brain derived neurophic factor (BDNF)and cyclic AMP (cAMP)by ELISA as well as the levels of cAMP-response element binding protein (CREB)and phosphorylation-cAMP-response element binding protein (pCREB)by western bolt. Results Compared with group S,the escape latency and swimming distance were significantly pro-longed in group T on day 1,3,7 and 14 and group A on day 1,3 after trauma (P <0.05 );while compared with group T,they were significantly shorter in group A on day 7,14 after trauma (P <0.05).Compared with group S,the levels of BDNF,cAMP,CREB and pCREB were significantly de-creased in group T(P < 0.05 ).Compared with group T,the levels of BDNF,cAMP,CREB and pCREB were significantly increased in group A (P <0.05).Conclusion SSA can significantly improve cognitive dysfunction in mice after traumatic brain injury,and the mechanism may be related to the activation of cAMP/CREB signaling pathway and up-regulation of BDNF.
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Objective To investigate the impact of enhanced recovery after surgery (ERAS)on immune function in patients undergoing laparoscopic hepatectomy.Methods Sixty patients undergoing laparoscopic hepatectomy (34 males,26 females,aged 38-57 years,ASA Ⅰ or Ⅱ),were randomly divided into two groups:enhanced recovery after surgery group (group E)and non-enhanced recovery after surgery group (group C).The patients in group E received enhanced recovery after surgery,while the patients in group C received routine perioperative management and anesthesia methods.The operation method and time,the volume of bleeding,the intraoperative fentanyl con-sumption,the volume of fluid input,the preoperative and postoperative CVP and temperature were recorded in the two groups.Blood samples were obtained before induction (T0 ),at the end of opera-tion (T1 ),on day 1 (T2 ),day 3 (T3 ),day 7 (T4 )after operation for determination of plasma con-centration of IgA,IgM,IgG and the percentages of T lymphocyte subsets (CD3+ ,CD4+ ,CD8+ ) and CD4+/CD8+ ratio were detected with flow cytometry.Furthermore,the visual analogue scale (VAS)score and Ramsay score were evaluated 4 hours,8 hours,24 hours and 48 hours after opera-tion in two groups.Results Compared with group C,the intraoperative fentanyl consumption,the volume of fluid input and the postoperative CVP in group E were significantly decreased,while the postoperative temperature was significantly increased (P < 0.05 ).Compared with T0 , the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ ratio and the plasma concentration of IgA,IgM,IgG in group E on T1-T3 were significantly decreased,the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ratio and the plasma concentration of IgA,IgM,IgG in group C on T1-T4 were significantly decreased (P <0.05).Compared with group C,the percentages of CD3+ ,CD4+ ,the CD4+/CD8+ ratio and the plasma concentration of IgA,IgM,IgG in group E on T1-T4 were significantly increased (P <0.05),the visual analogue scale (VAS)score 4 hours,8 hours,24 hours after operation were signifi-cantly decreased (P <0.05).The comparision of Ramsay scores at all the time points between two groups were similar.Conclusion ERAS applied to patients undergoing laparoscopic hepatectomy can reduce the intraoperative fentanyl consumption,prevent the occurrence of hypothermia and provide satisfactory postoperative analgesia,which can significantly improve the immune function in patients.
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Objective To evaluate the effect of stellate ganglion block (SGB) on cellular immune function in diabetic rats.Methods Healthy male Sprague-Dawley rats,aged 3 months,weighing 240-280 g,were used in this study.Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.Forty-eight rats with diabetes mellitus were randomly divided into 2 groups (n=24 each) using a random number table:diabetes mellitus group (group DM) and group SGB.Another 24 healthy rats,aged 3 months,were selected and served as control group (group C).At 1 week after successful establishment of the model,unilateral transection of cervical sympathetic trunk (TCST) was performed in group SGB,while the right cervical sympathetic trunk was only exposed in C and DM groups.Before TCST (T0) and on 1,3,7 days after TCST (T1-3),6 rats were randomly selected from each group,and blood samples were collected from the inferior vena cava for determination of the blood glucose,plasma norepinephrine (NE) concentrations (by enzyme-linked immunosorbent assay),and levels of T lymphocyte subsets CD3+,CD4+ and CD8+ in whole blood (using FACSCalibur flow cytometer).C D4+/CD8+ratio was calculated.The rats were weighed before sacrifice,and the rats were sacrificed to obtain the thymus which was weighed.The thymus index (thymus weight/body weight) was calculated.Results Compared with group C,the blood glucose was significantly increased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly decreased at T0-3 (P＜0.05),and no significant change was found in CD8+ levels in DM and SGB groups (P＞0.05),the plasma NE concentrations were significantly decreased at T1-3 in group SGB (P＜0.05),and no significant change was found in plasma NE concentrations in group DM (P＞0.05).Compared with group DM,the blood glucose and plasma NE concentrations were significantly decreased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly increased at T1-3 (P＜0.05),and no significant change was found in CD8+ levels in group SGB (P＞0.05).Conclusion SGB can improve the cellular immune function in diabetic rats.
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Objective To evaluate the effect of perioperative administration of dexmedetomidine on postoperative ileus after laparocolectomy.Methods Sixty patients scheduled for abdominal surgery were randomly divided into two groups,30 in each group.Group D received dexme-detomidine administeration at a loading dose of 0.6 μg/kg for 10 minutes before induction,followed by an infusion rate of 0.5 μg·kg-1 ·h-1 to 30 min before the end of surgery.The control group re-ceived saline instead of Dex.After the surgery,Group C received intravenous sufentanyl 2 μg/kg, while group D sufentanyl 2 μg/kg combined with Dex 2 μg/kg.Heart rate variability (HRV)were detected before Dex infusion (T0 ),10 minutes after intubation (T1 ), 10 minutes after CO 2 insufflation (T2 ),1 hour after CO 2 insufflation (T3 ),10 minutes after CO 2 desufflation (T4 ),and 10 minutes after extubation(T5 ).The plasma concentrations of epinephrine(E)and norepinephrine (NE)were determined at T0 ,T3 ,T5 ,T7 and T1 0.The recovery of bowel function was evaluated in terms of the first time to fart and intake food.Results Compared with T0 ,HRV of power (TP), high-frequency (HF)power,low-frequency (LF)power and the ratio of LF/HF power were signifi-cantly decreased at T1-T4 in group C and at T1-T5 in group D.The plasma concentrations of E and NE were higher at T3 ,T5 ,T7 and T1 0 in both group C and group D (P <0.05).Compared with T1 ,TP, LF and the ratio of LF/HF were increased at T2-T4 (P <0.05).Compared with group C,TP,LF and the ratio of LF/HF were decreased at T2-T5 ,The plasma concentrations of E and NE were decreased at T3 ,T5 ,T7 and T10 and the time of first flatus was earlier(P <0.05).Conclusion The perioperative ad-ministration of dexmedetomidine during laparocolectomy facilitated the early recovery of bowel func-tion after surgery and decreasede the time of postoperative ileus.
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Objective: To investigate the effect of sevolfurane (SEVO) post-conditioning on protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway for protecting ischemia/reperfusion (I/R) injury in isolated rat’s heart. Methods: A total of 84 isolated rat’s heart prepared by Langendorff method were randomly divided into 7 groups andn=12 in each group.①Sham group,②I/R group,③SEVO post- conditioning (SPC) group,④NVP-BEZ235 solvent dimenthyl sulfoxide (DMSO) group,⑤Phosphatidyl inositol 3-kinase (PI3K)/mTOR dual inhibitor NVP-BEZ235 (BEZ) group,⑥BEZ+SPC group and⑦SEVO alone group. The hearts received 30 min ischemia followed by 120 min reperfusion with relevant treatment except for Sham group and SEVO group in which the hearts were without ischemia process. Myocardial infarct (MI) size and tissue pathological changes were observed, protein expressions of phosphor-AKT (P-AKT)/total-AKT, P-mTOR/total-mTOR, Beclin1, Bax/Bcl-2 and cleaved Caspase-3 were examined at the end of reperfusion respectively. Results: Compared with I/R group, SPC group presented decreased MI size (26.28±4.00) % vs (49.22±3.66) % and reduced tissue pathological changes; up-regulated protein expressions of P-AKT/total-AKT and P-mTOR/total-mTOR by 79.85% and 67.02%, while down-regulated protein expressions of Beclin1, Bax/Bcl-2 and cleaved Caspase-3 by 33.77%, 69.26% and 48.84%respectively, allP<0.05. Compared with SPC group, BEZ+SPC group sowed increased MI size (53.85±4.06) % vs (26.28±4.00) %and elevated tissue pathological changes; down-regulated proteins expressions of P-AKT/total-AKT and P-mTOR/total-mTOR by 46.06% and 42.95%, while up-regulated protein expressions of Beclin1, Bax/Bcl-2 and cleaved Caspase-3 by 29.90%, 206.85% and 114.65% respectively, allP<0.05. Conclusion: SPC may activate AKT/mTOR pathway and inhibit cardiomyocyte autophagy and apoptosis, thereby attenuate I/R injury in isolated rats’ heart.
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<p><b>OBJECTIVE</b>To observe the effect of electroacupuncture (EA) combined with general anesthesia for the immune function of patients treated with laparoscopic radical rectectomy for rectal cancer.</p><p><b>METHODS</b>Fifty patients who would receive selective laparoscopic radical rectectomy for rectal cancer with general anesthesia were randomly divided into an observation group and a control group,25 cases in each one. Fifteen minutes before anesthesia induction,patients in the observation group were treated with EA at Zusanli (ST 36) and Sanyinjiao (SP 6) until the end of operation. Sham acupuncture without piercing the skin was applied at the same acupoints in the control group, and electrodes were connected without stimulation. Interferon-γ (IFN-γ), interleukin-4 (IL-4), interleukin-6 (IL-6) were quantitatively tested before anesthesia (T), at the time of abdomen closing (T) and one hour after anesthesia anabiosis (T). And serum procalcitonin (PCT) level, leucocyte count and the number of cases with increasing leucocyte (the standard number>10×10/L) were measured on the first day after operation.</p><p><b>RESULTS</b>The levels of IL-4 and IL-6 were increased apparently and the ratio of IFN-γ/IL-4 was decreased at Tcompared with those before treatment in the control group (all<0.05), but obvious change did not appear in the observation group (all>0.05). The ratio of IFN-γ/IL-4 was enhanced (<0.05),and the levels of IL-4 and IL-6 were reduced (both<0.05) at Tin the observation group compared with those in the control group. The level of PCT of the observation group was markedly lower than that of the control group on the first day after operation (<0.05). There was no statistical significance about leucocyte count and the number of cases with increasing leucocyte between the two groups (both>0.05).</p><p><b>CONCLUSIONS</b>EA at Zusanli (ST 36) and Sanyinjiao (SP 6) could alleviate the depressing immune function and inflammatory reaction of patients after laparoscopic radical rectectomy for rectal cancer.</p>
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Objective To investigate the role of AKT/GSK3β/mTOR signaling molecule in myocardial protection of sevoflurane postconditioning.Methods Thirty-nine male Sprague-Dawley rats,weighing 200-250 g,installed in vivo myocardial ischemia-reperfusion model by left anterior de-scending coronary occlusion for 30 min.Rat hearts were randomly divided into 3 groups (n = 13 ):sham control group (group Sham),purely ischemia-reperfusion group (group IR),sevoflurane post-conditioning group (group SPC).With the exception of group Sham,each group was subjected to oc-clusion for 30 min followed by 2 h reperfusion.Group SPC was subjected to sevoflurane postcondi-tioning:2.4% sevoflurane was inhaled for 1 5 min starting from the end of ischemia until 1 5 min after beginning of reperfusion,while 33% oxygen was inhaled in the other groups.At the end of 2 h reper-fusion,cardiac function was evaluated by two-dimensional echocardiography,myocardial infarction size was measured by using 1% 2,3,5-triphenyltetrazolium chloride triazole (TTC),myocardial ultra-structural alterations was detected by transmission electron microscopy (TEM),cardiomyocytes ap-optosis was examined by terminal deoxynucleotidyl nickend labeling (TUNEL),the expressions of p-AKT/t-AKT, p-GSK3β/t-GSK3β, p-mTOR/t-mTOR,Bcl-2 and Bax proteins was measured by Western blot.Results Compared with group Sham,cardiac function was deteriorated,myocardial in-farct size was increased,cardiomyocyte mitochondrial damage was increased,positive apoptotic car-diomyocyte was increased,the expression of Bcl-2 was down-regluated,and the expressions of p-AKT/t-AKT,p-GSK3β/t-GSK3β,p-mTOR/t-mTOR and Bax were up-regluated in group IR (P <0.05).Compared with group IR,cardiac function was improved,myocardial infarct size was de-creased,cardiomyocyte mitochondrial damage was decreased,positive apoptotic cardiomyocyte was decreased,the expression of Bax was down-regluated,and the expressions of p-AKT/t-AKT,p-GSK3β/t-GSK3β,p-mTOR/t-mTOR and Bcl-2 were up-regluated in group SPC (P < 0.05 ). Conclusion Sevoflurane postconditioning can mitigate ischemia-reperfusion injury to in vivo rat hearts,decreased cardiomyocyte mitochondrial damage,inhibited cardiomyocyte apoptosis,and its mechanism was related to the activation of AKT/GSK3β/mTOR signaling molecule.