Penetration ability and biological activity of fusion protein PTD4-Cu, Zn-SOD in human astrocytes in vitro by immunofluorescence test / 西安交通大学学报(医学版)

【Objective】 To observe the penetration and biological activity of PTD4-Cu, Zn-SOD into human astrocytes and whether it can mitigate hypoxia damages. 【Methods】 ①Immunohistochemistry and fluorescence test: We labeled the Cu, Zn-SOD by a monoclonal antibody, combined it with the fluorescent secondary antibody labeled with fluorescein isothiocyanate (FITC) to observe the effect of transduced PTD4-Cu, Zn-SOD on the viability of human astrocytes. ② The experimental group: After hypoxic damage model, the cells were divided into three groups: blank control, group Cu, Zn-SOD, and group PTD4-Cu, Zn-SOD. Group blank was added with DMEM medium (excluding serum) as control; DMEM medium was added to the other two for one hour (excluding serum) with its fusion proteins (Cu, Zn-SOD and PTD4 -Cu, Zn-SOD) with the final concentration of 2 μmoL/L. After the intervention, we used SOD and MDA test kits to observe PTD4-Cu, Zn-SOD and Cu, Zn-SOD in astrocytes after fusion protein intervention. 【Results】 The PTD4-Cu, Zn-SOD fusion protein could have aggregation distribution in the nucleus by FITC fluorescently labeled. After the intervention, it could increase the SOD activity in astrocytes in group PTD4-Cu, Zn-SOD and group Cu, Zn-SOD compared with control group, but the SOD activity was more obvious in the fusion proteins PTD4-Cu, Zn-SOD group. And the dose of MDA was reduced in group PTD4-Cu, Zn-SOD compared with group Cu, Zn-SOD and control group. 【Conclusion】 PTD4-Cu, Zn-SOD fusion protein can transcellular membrane of human astrocytes. The fusion protein PTD4-Cu, Zn-SOD can increase the SOD activity and reduce the content of MDA by human astrocytes from hypoxia injury.

Efficacy of dexmedetomidine on painless gastroenteroscopy in patients recovered from COVID-19 / 中华麻醉学杂志

Objective:To evaluate the efficacy of dexmedetomidine on painless gastroenteroscopy in the patients recovered from COVID-19.Methods:Eighty patients of either sex, aged 18-64 yr, with body mass index <30 kg/m 2, undergoing elective painless gastroenteroscopy, of American Society of Anesthesiologists physical statusⅠor Ⅱ, within 2nd to 7th weeks after diagnosis of COVID-19, who had a negative nucleic acid test or antigen test at present and presented with no respiratory symptoms in our hospital from January to February 2023, were selected and divided into 2 groups ( n=40 each) using a random number table method: dexmedetomidine group (group D) and control group (group C). Sufentanil 0.1 μg/kg was intravenously injected in two groups. Dexmedetomidine 0.2 μg/kg was intravenously injected in group D, and the equal volume of normal saline was given instead in group C. Propofol 1.5-2.0 mg/kg was intravenously injected after 2-3 min observation, and propofol 5-7 mg·kg -1·h -1 was intravenously infused to maintain sedation during operation. The development of bucking and hypoxemia during operation, total consumption of propofol, emergence time, time of hospital discharge, development of bradycardia and hypotension during operation, and scores for patients′ and endoscopic physicians′ satisfaction with anesthesia were recorded. Results:Compared with group C, the incidence of bucking and hypoxemia was significantly decreased, scores for endoscopic physicians′ satisfaction with anesthesia were increased ( P<0.05), and no significant change was found in the other parameters in group D ( P>0.05). Conclusions:Low-dose dexmedetomidine can reduce the risk of bucking and hypoxemia during operation and raise the quality of anesthesia in the patients recovered from COVID-19 undergoing painless gastroenteroscopy.

Effect of etomidate and dexamethasone on cortisol secretion in elderly patients undergoing general anesthesia / 中华麻醉学杂志

Objective:To evaluate the effects of dexamethasone and etomidate on cortisol secretion in elderly patients undergoing general anesthesia.Methods:One hundred and twenty-five elderly patients of either sex, aged 66-90 yr, of American Society of Anesthesiologists physical statusⅠ-Ⅲ, undergoing minor and medium elective surgeries under general anesthesia, were allocated into 4 groups using a random number table method: propofol and normal saline group (group PN, n=31), propofol and etomidate group (group PD, n=31), etomidate and normal saline group (group EN, n=33) and etomidate and dexamethasone group (group ED, n=30). In PN and EN groups, propofol (2 mg/kg) was used to induce and maintain anesthesia, and normal saline 2 ml and dexamethasone 0.1 mg/kg were intravenously injected, respectively, at 5 min before anesthesia induction.In PD and ED groups, etomidate (0.2 mg/kg) was used to induce and maintain anesthesia, and normal saline 2 ml and dexamethasone 0.1 mg/kg were intravenously injected, respectively, at 5 min before anesthesia induction.The serum cortisol concentrations were measured at 8: 00 after entering the operating room on the morning of operation (T 1), 1 h after the start of anesthesia (T 2), 2 h after the start of anesthesia (T 3), 8: 00 on the next day ofoperation (T 4) and 8: 00 on the 2nd day of operation (T 5). Blood glucose concentrations were measured at T 1-T 3, and the hypotension during the peri-anesthesia period, nausea and vomiting in post-anesthesia care unit, and nausea and vomiting scores were recorded at 24 h after operation. Results:A total of 122 patients completed the trial.Compared with PN group, the concentration of serum cortisol was significantly decreased at T 2-T 5, blood glucose concentrations were increased at T 2 and T 3 ( P<0.05), and no significant change was found in the incidence of hypotension, nausea and vomiting and nausea and vomiting scores in PD group ( P>0.05), and the concentration of serum cortisol was significantly decreased at T 2-T 4, the incidence of hypotension was decreased ( P<0.05), and no significant change was found in the blood glucose concentrations, incidence of nausea and vomiting or nausea and vomiting scores in EN group ( P>0.05). Compared with ED group, the serum cortisol concentration was significantly increased at T 2 and T 3, the incidence of hypotension was increased, the incidence of nausea and vomiting and nausea and vomiting scores were decreased ( P<0.05), and no significant change was found in the blood glucose concentrations in PD group ( P>0.05), and the serum cortisol concentration was significantly decreased at T 2 and T 3 and increased at T 4 and T 5, the serum cortisol concentration was decreased at T 2 and T 3, and no significant change was found in the incidence of hypotension, nausea and vomiting and nausea and vomiting scores in EN group ( P>0.05). Conclusions:Combination of etomidate and dexamethasone significantly enhances the duration and degree of inhibition of cortisol secretion in elderly patients than etomidate or dexamethasone alone.

Effects of different depths of anesthesia on CD4 + T cell function in patients undergoing radical resection for malignant tumor / 中华麻醉学杂志

Objective:To evaluate the effects of different depths of anesthesia on CD4 + T cell function in the patients undergoing radical resection for malignant tumor. Methods:Forty-two American Society of Anesthesiologists physical status Ⅱ patients, aged 36-64 yr, weighing 49-95 kg, undergoing elective radical surgery for malignant tumor, were divided into 2 groups ( n=21 each) using a random number table method: light anesthesia group (L group) and deep anesthesia group (D group). The AAI values were maintained at 30-40 and 20-29 during operation in L group and D group, respectively.The time to eye opening, extubation time and consumption of propofol were recorded.The peripheral venous blood samples were collected immediately before induction of anesthesia (T 0), at 2 h of operation (T 1) and at 4, 24, and 72 h after operation (T 2-4) for determination of serum interferon (IFN)-γ and interleukin-4 (IL-4) concentrations (by enzyme-linked immunosorbent assay), and IFN-γ/IL-4 ratio was calculated. Results:Compared with the baseline value at T 0, the serum IFN-γ concentrations were significantly increased at T 3, 4, the serum IL-4 concentrations were decreased at T 1-4, and IFN-γ /IL-4 ratio was increased in group D ( P<0.05). Compared with group L, the serum IFN-γ concentrations were significantly increased at T 3, 4, the serum IL-4 concentrations were decreased at T 1-4, IFN-γ/IL-4 ratio was increased, the consumption of propofol was increased, and the extubation time was prolonged in group D ( P<0.05), and no significant change was found in the time to eye opening between the two groups ( P>0.05). Conclusion:Deep anesthesia can improve CD4 + T cell function in the patients undergoing radical resection for malignant tumor.

Efficacy of oxycodone for improvement of general anesthesia for laparoscopic cholecystectomy in elderly patients / 中华麻醉学杂志

Objective To evaluate the efficacy of oxycodone for improvement of general anesthesia for laparoscopic cholecystectomy in elderly patients.Methods A total of 160 patients of both sexes,aged 65-75 yr,with body mass index ＜30 kg/m2,of American Society of Anesthesiologists physical status Ⅰ-Ⅲ,scheduled for elective laparoscopic cholecystectomy,were divided into 2 groups (n =80 each) using a random number table method:general anesthesia group (group GA) and oxycodone + general anesthesia group (group OX+GA).Anesthesia induction:propofol was given by closed-loop infusion at the initial target plasma concentration of 2 μg/ml,the target bispectral index (BIS) value was set at 50,and 2 min later remifentanil was given by target-controlled infusion at the target plasma concentration of 4 ng/ml,and cisatracurium 0.2 mg/kg was intravenously injected when BIS value was decreased to 70.Laryngeal mask airways were inserted and the patients were mechanically ventilated when BIS value was decreased to 50 and TOF ratio was decreased to 25％,and end-tidal pressure of carbon dioxide was maintained at 35-45 mmHg.Anesthesia maintenance:propofol was given by closed-loop infusion,the target BIS value was set at 50,cisatracurium 0.1 mg/kg was intravenously injected when TOF ratio was increased to 10％;remifentanil was given by target-controlled infusion at the target plasma concentration of 4-6 ng/ml.Oxycodone 0.07 mg/kg was intravenously injected at 5 min before stretching internal organs.Before anesthesia,at 5 min after laryngeal mask airway placement,at skin incision and while stretching internal organs,analgesia nociception index value and perfusion index value were recorded,the development of intraoperative cardiovascular events,emergence time,time for removal of laryngeal mask airway,time of post-anesthesia care unit stay and development of nausea and vomiting and back and shoulder pain within 48 h after surgery were also recorded.Results Compared with group GA,the analgesia nociception index value and perfusion index value were significantly increased while stretching internal organs,and the incidence of intraoperative hypertension,tachycardia,and nausea and vomiting and back and shoulder pain within 48 h after surgery were decreased in group OX-GA (P＜0.05).Conclusion Oxycodone can inhibit nociceptive stimuli,is helpful in maintaining stable hemodynamics and reduces postoperative complications in elderly patients undergoing laparoscopic cholecystectomy under combined general anesthesia.

Preparation of PTD4-Cu,Zn-SOD fusion protein / 中华麻醉学杂志

Objective To prepare PTD4-Cu,Zn-SOD fusion protein.Methods The recombinant plasmid of pET 1 6b-Cu,Zn-SOD and pET16b-PTD4-Cu,Zn-SOD was transformed into Escherichia coli BL21 (DE3).Isopropyl β-D-1-thiogalactopyranoside was then added at a final concentration of 0.84 mmol/L,and the cells were incubated for 4 h to induce the expression of Cu,Zn-SOD and PTD4-Cu,Zn-SOD fusion protein.Lysozyme and ultrasound were used to lyse the bacteria,the supernatant was collected for 15％ SDS-PAGE to analyze the expression of the target protein.Ni-NTA His bind resin was used to purify Cu,Zn-SOD protein and PTD4-Cu,Zn-SOD fusion protein under natural conditions.Western blot was used to identify the target protein.Results The results of Western blot showed that the purity of the target protein was about 90％,and the Cu,Zn-SOD protein with a molecular weight about 19 kDa and the PTD4-Cu,Zn-SOD fusion protein with a molecular weight about 20 kDa were found.Conclusion PTD4-Cu,Zn-SOD fusion protein is prepared successfully.

Effect of theanine pretreatment on DNA repair function in neurons during brain ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To investigate the effect of theanine pretreatment on DNA repair function in neurons during brain ischemia-reperfusion (I/R) injury in rats.Methods One hundred and eight male Sprague-Dawley rats,weighing 290-310 g,aged 15 weeks,were randomly divided into 3 groups (n =36 each) using a random number table:sham operation group (S group),I/R group and theanine pretreatment group (T group).Global cerebral I/R was produced by 4-vessel occlusion method.Bilateral vertebral arteries were electrically cauterized,and bilateral common carotid arteries were clamped for 6 min.Theanine 1 g/kg was injected intravenously at 4 h before clamping bilateral common carotid arteries in T group,and the equal volume of normal saline was given in the other two groups.At 2,6,12,24,48 and 72 h of reperfusion,6 rats were selected in each group and sacrificed,the brains were removed,and the hippocampus was isolated for determination of the number of viable neurons in the hippocampal CA1 region (with a light microscope),apoptosis in neurons in the hippocampal CA1 region (by TUNEL),and expression of DNA repair protein X-ray repair cross-complementing group 1 (XRCC1) and Ku70 (by immunohistochemistry).The apoptotic index was calculated.Results Compared with S group,the number of viable neurons was significantly decreased,and the apoptotic index was significantly increased at 6,12,24,48 and 72 h of reperfusion,and the expression of XRCC1 and Ku70 was significantly down-regulated at 2,6,12,24,48 and 72 h of reperfusion in I/R group (P＜0.01).Compared with I/R group,the number of viable neurons was significantly increased at 12,24,48 and 72 h of reperfusion,the apoptotic index was significantly decreased at 6,12,24,48 and 72 h of reperfusion,and the expression of XRCC1 and Ku70 was significantly up-regulated at 2,6,12,24,48 and 72 h of reperfusion in T group (P ＜ 0.01).Conclusion The mechanism by which theanine pretreatment attenuates brain I/R injury is related to enhancement of DNA repair function and reduction of neuronal apoptosis in rats.

Construction of prokaryotic recombinant expression vector of PTD4-Cu, Zn-SOD / 中华麻醉学杂志

Objective To construct the prokaryotic recombinant expression vector of PTD4-Cu,Zn-SOD.Methods By using the techniques of gene recombination,the primers of Cu,Zn-SOD and the oligonucleotide sequences of PTD4 were designed,PCR amplification was performed for Cu,Zn-SOD genes,the PCR products were identified,reclaimed and purified,and pET16b served as carrier.The prokaryotic recombinant expression vector of pET16b-Cu,Zn-SOD was constructed using double digestion with Xho Ⅰ and BamH Ⅰ,ligated reaction and plasmid transformation.Then PTD4 gene and pET16b-Cu,Zn-SOD carrier were double digested with Nde Ⅰ and Xho Ⅰ and ligated,and the plasmid was transformed,and the prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD was constructed.The reconstructed vector was analyzed by restriction mapping and was verified by gene sequencing.Results The prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD with a length of 6 207 bp was constructed successfully.The carrier fragment about 5.7 kp and PTD4-Cu,Zn-SOD gene fragment about 510 bp were obtained by double digestion with Nde Ⅰ and BamH Ⅰ,which was consistent with the expected results.The results of gene sequencing showed that the base sequences of pET16b-PTD4-Cu,Zn-SOD were correct when compared with the expected gene sequences.Conclusion The prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD is constructed successfully.

Influence of Propofol and Etomidate on Cognitive Dysfunction of Elderly Patients after Laparoscopic Chole-cystectomy / 中国药房

OBJECTIVE:To discuss the effect of etomidate and propofol on early postoperative cognitive dysfunction (POCD)of elderly patients after laparoscopic cholecystectomy(LC)and significance of serum protein S100β to the occurrence of early POCD in total intravenous anesthesia. METHODS:60 patients aged 65 years old above undergoing LC in total LMA intrave-nous anesthesia were selected and randomly divided into etomidate group(group E)and propofol group(group P),with 30 cases in each group. Anesthesia was induced by etomidate 0.3 mg/kg (group E) or propofol 1.5 mg/kg (group P),and additionally in-duced by sufentanil 0.4 μg/kg and vecuronium 0.12 mg/kg. Anesthesia was maintained with intravenous pump of remifentanil 0.15 μg/(kg·min),continuous target controlled infusion of etomidate(target concentration 1.0-1.5 μg/ml)(group E)or propofol(target concentration 3.0-4.0 μg/ml)(group P);the dual brain index(BIS)values were maintained between 40 and 50 throμgh adjusting target concentration of etomidate or propofol. The blood samples were collected 1 h before operation(T0),2 h(T1),24 h(T2), 48 h(T3)after operation,and the content of S100βprotein was detected and mini-mental state examination(MMSE)score were re-corded. Meanwhile,recovery time,laryngeal mask removal time,intraoperative dosage and the occurrence of intraoperative aware-ness were observed and recorded in 2 groups. RESULTS:There was no statistically significant difference in MMSE score between 2 groups at different time points(P>0.05);MMSE score of 2 groups at T1 and T2 was significantly lower than at T0,with statisti-cal significance(P0.05);The contents of S100β protein in 2 groups at T1 and T2 were significantly higher than at T0,with sta-tistical significance(P0.05). The amount of ephedrine in group P was significantly higher than in group E,with statisti-cal significance (P<0.05). No intraoperative awareness oc-curred in 2 groups throμgh postoperative follow-up. CONCLUSIONS:Etomidate and propofol total intravenous anesthesia can be safely used in elderly patients with LC,and they can cause short-term POCD at different degrees. The amount of S100β protein has some relevance with the occurrence of early POCD .

A placebo-controlled clinical trial to evaluate the efficacy and safety of domestic palonosetron hydrochloride injection on the prevention of postoperative nausea and vomiting / 临床麻醉学杂志

Objective To evaluate the efficacy and safety of domestic palonosetron hydrochlo-ride injection on its prevention of postoperative nausea and vomiting.Methods A multi-centered,ran-domized,double-blinded and placebo-controlled clinical trial was carried out.A total of 281 patients were enrolled,with 141 of patients in study group and 140 of patients in control group respectively. 0.075 mg of intravenous palonosetron hydrochloride injection was delivered in the study group before anesthesia induction.The drug was substituted by 1.5 ml of NS in the control group.All anesthesia inductions were conducted by the intravenous injection of propofol,fentanyl and rocuronium,and were maintained with sevoflurane and fentanyl.Complete remission rate and treatment failure cut-off time of vomiting were evaluated at 0-6 h,6-72 h,0-72 h postoperatively.Results In the study group CR% 0-6 h,6-72 h and 0-72 h were 107 (75.89%),104 (73.76%)and 92 (65.25%),the control group was 81 (57.86%),70 (50%)and 62 (42.86%),CR% of the study group was significantly higher than that of the control group (P <0.01).Insignificant statistical difference but significant clin-ical difference exists in their treatment failure cut-off time,386.5 min and 300.0 min,respectively be-tween the groups.Conclusion Domestic palonosetron hydrochloride injection is safe and effective in the prevention of postoperative nausea and vomiting.

Effect of dexmedetomidine on stress responses during wake-up test in patients undergoing cerebral functional area operation performed under propofol combined with remifentanil anesthesia / 中华麻醉学杂志

Objective To investigate the effect of dexmedetomidine on the stress responses during wakeup test in patients undergoing cerebral functional area operation performed under propofol combined with remifentanil anesthesia.Methods Thirty-six ASA physical status Ⅰ or Ⅱ patients,undergoing cerebral functional area operation requiring wake-up test,aged 18-60 yr,weighing 50-70kg,were randomly divided into control group (group C) or dexmedetomidine group (group D) with 18 cases in each group.Dexmedetomidine 0.8 μg/kg was infused over 10 min before induction of anesthesia followed by infusion at 0.4 μg·kg-1 · h-1 in group D,while the equal volume of normal saline was infused in group C.Anesthesia was induced with target-controlled infusion of propofol and remifentanil and iv injection of cisatracurium.At 30 min prior to wake-up test,target-controlled infusion of propofol and application of mulscle relaxants were stopped,the target plasma concentration of remifentanil was decreased to 1 ng/ml,and in group D the infusion rate of dexmedetomidine was decreased to 0.1 μg·kg 1· h-1.Anesthesia time and consumption of anesthetics before wake-up test,wake-up time,and development of complications and intraoperative awareness during wake-up test were recorded.At 30 min prior to wake-up test (T1),immediately after wake-up (T2),at 5 min after wake-up (T3),and at 10 min after the anesthetic depth was deepened (T4),HR,mean arterial pressure and BIS value were recorded and blood samples were taken for determination of plasma concentrations of epinephrine (E) and norepinephrine (NE).Results Compared with group C,the consumption of propofol and remifentanil was significantly reduced before wake-up,the incidence of hypertension was decreased during wake-up test,and HR and plasma E and NE concentrations were decreased at each time point (P ＜ 0.05),and no significant difference in mean arterial pressure and BIS value was found in group D (P ＞ 0.05).Tachycardia,restlessness,bucking and awareness were not observed during wake-up test in group D.Conclusion Dexmedetomidine can inhibit the stress responses during wake-up test and raise the quality of wake-up test in patients undergoing cerebral functional area operation performed under propofol combined with remifentanil anesthesia.

Effect of acute normovolemic hemodilution on apoptosis in hippocampal cells induced by global cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

ObjectiveTo investigate the effect of acute normovolemic hemodilution (ANH) on the apoptosis in hippocampal cells induced by global cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy 50-60 day old male SD rats weighing 280-320 g were randomly divided into 3 groups ( n =12 each):group sham operation (group S); group global cerebral I/R (group I/R) and group ANH.Global cerebral I/R was produced by 4-vessel technique described by Pulsinelli et al.in groups I/R and ANH.ANH was carried out at 24 h after cauterization of bilateral vertebral arteries,before occlusion of bilateral carotid arteries.Blood was withdrawn from femoral artery until Hct was reduced to 30％ and equal volume of hydroxyethyl starch 130/0.4 sodium chloride was infused into femoral vein simultaneously.Bilateral carotid arteries were blocked for 5 min at 10 min after ANH.The rats were sacrificed at 24 h of reperfusion and their hippocampi were isolated.Apoptosis was detected by flow cytometry.The expression of Apaf-1 mRNA and caspase-3 mRNA was determined by RT-PCR.Results Global cerebral I/R significantly increased apoptosis index and up-regulated Apaf-1 mRNA and caspase-3 mRNA expression in group I/R as compared with group S.ANH significantly attenuated apoptosis and down-regulated Apaf-1 mRNA and caspase-3 mRNA expression in group ANH compared with group I/R.ConclusionANH can reduce hippocampal cell apoptosis induced by cerebral I/R through down-regulation of Apaf-1 and caspase-3 expression in hippocampus.

Effect of tea polyphenols on global cerebral ischemia reperfusion injury in rats / 中华麻醉学杂志

Objective To investigate the effect of tea polyphenols on global cerebral ischemia reperfusion injury in rats.Method Forty-five pathogen-free male SD rats weighing 180-220 g were randomly divided into 3 groups( n =15 each):sham operation group (group S),cerebral ischemia reperfusion group (group IR) and tea polyphenols group (group TP).Global cerebral ischemia reperfusion injury was establish by four-vessel occlusion method.At 24 h of reperfusion,five rats were chosen and Evan's blue(EB) was injected iv,and then sacrificed and brain was removed for determination of EB content; another five rats were sacrificed and brain was removed for determination of water content; five rats were chosen for Morris water maze test.Result Compared with group S,EB content and water content in brain tissue were increased in groups IR and 'rP,and escape latency was prolonged,frequency of crossing the original platform was reduced in group IR ( P ＜ 0.05 ).Compared with group IR,EB content and water content in brain tissue were decreased,and escape latency was shortened,frequency of crossing the original platform was increased in group Tp ( P ＜ 0.05).Conclusion Tea polyphenols can attenuate global cerebral ischemia reperfusion injury in rats.

Role of extracellular signal-regulated kinase in neuronal apoptosis of hippocampns induced by global cerebral ischemia-reperfusion in rats / 中华麻醉学杂志

Objective To investigate the role of extracellular signal-regulated kinase in neuronal apoptosis of hippoeampus induced by global cerebral ischemia-reperfusion in rats.Methods Ninety healthy male SD rats weighing 280-320 g were randomly divided into 3 groups(n=30 each):groupI sham operation(S);groupⅡI/ R and group Ⅲ PD98059+I/R(PD).The animals were anesthetized with intraperitoneal 1% pentobarbital 40 mgkg.Global cerebral I/R was produced by 4-vessel occlusion method.Bilateral vertebral arteries were electrically cauterized and bilateral common carotid arteries were clamped for 5 min.Clamping was then released for reperfusion in group Ⅱ and Ⅲ.In group Ⅲ PD98059(a specific ERK inhibitor)O.3 mg/kg was injected iv before carotid artery clamping.Five animals in each group were sacrificed at 2,6,12,24,48 and 72 h of reperfusion and their brains were removed and cut into sections which were stained with HE and examined under microscope(400 times magnified).Neuronal apoptosis in hippieampal Cal and CA3 regions were detected by TUNEL assay.Apoptotic index(AI) was calculated.Phosphorylated ERK(p-ERK)and phosphorylated Bad (p-Bed)expression was assessed by immuno-histochemistry.Results (1)Hippoeampal pyramidal cells were orderly distributed and morphologically intact in group S but unevenly distributed with widened intercellular space,shrinked cell body and condensed nuclei in I/R group.The I/R induced changes were worse in PD group.(2)AI in Cal and CA3 region was significantly higher in PD group than in I/R group.(3)The p-ERK expression in Cal region was signiticanfly decreased at 2,6,12 h of reperfusion in I/R(Ⅲ)and PD groups(Ⅲ)as compared with group S(I),while p-ERK expression in CA3 region was significantly lower at 2,6,12,24 h of reperfusion in PD group(Ⅲ)than in S(I)and I/R group(Ⅱ).(4)The p-Bed expression in Cal and CA3 regio was signi ficantly lower during reperfusion in PD group than in I/R group and was lowest in group S(I).Conclusion Cerebral isehemia reperfusion can decrease the p-ERK expression and then leads to the dephosphorylation of Bed(a member of Bel-2 protein family),which induced the apeptosis in hippoeampal neurons.

Protective effects of fructose 1,6-diphosphote on the brain against ischemia-reperfusion injury in rats / 中华麻醉学杂志

Objective To investigate the protective effect of fructose 1,6-diphosphate (FDP) on the brain against ischemia-reperfusion (I/R) injury and the possible mechanism. Methods One hundred and eighty SD rats weighing 275-325 g were randomly divided into 3 groups (n = 60 each): Ⅰ gham operation group; Ⅱ I/R group and Ⅲ FDP group. Global cerebral I/R was produced by 4-vessel technique. Bilateral vertebral arteries were coagulated and bilateral common carotid arteries were occluded for 5 min and then released for reperfusion. In sham operation the four vessels were exposed but not occluded. In FDP group FDP 1.5 mg?kg-1 was given Ⅳ when reperfusion was started, while in sham-operation group and I/R group normal saline (NS) 1.5 ml?kg-1 was given Ⅳ instead of FDP. The animals were killed at 2, 6, 12, 24, 48 and 72 h of reperfusion ( n = 5 each) for determination of cerebral SOD activity and MDA contents, the number of apoptotic neurons (TUNEL) and expression of P38 and Ref-1 in the brain (immuno-histochemical method) .Results The MDA content was significantly higher whereas the SOD activity and P38 and Ref-1 expression were significantly lower at all time points in I/R group than in sham operation group ( P

Protective effects of propofol against ischemia-reperfusion injury in rat brain / 中华麻醉学杂志

Objective To study the protective effects of propofol against ischemia-reperfusion injury in rat brains.Methods Modified Longa modle of focal cerebral ischemia-reperfusion injury was used. 200 healthy male SD rats, weighing 200-300g were anesthetized with intraperitoneal(I.P.) ketamine and propofol. When righting reflx was abolished, external carotid artery was exposed. A nylon thread with rounded end was inserted cranially until anterior cerebral artery was reached. After 3h ischemia nylon thread was withdrawn for reperfusion which lasted 3h. Bloos samples were obtained from orbit. Skull was opened and brain removed. In control group carotid artery was exposed but nylon thread was not inserted cranially. The animals were divided into four groups: (1)ischemia-reperfusion model group: normal saline 10 ml was administered I.P.,(2)operation control group: normal saline was given I.P.at the end of operation,(3)nimodipine group: nimodipine 1 mg?kg -1 was administered I.P. 10 min before ischemia,(4) propofol group: propofol 110 mg?kg -1 was given I.P. 10 min before ischemia. Brain infarction area, cerebral water content, serum lactate dehydrogenase(LDH) and creatine kinase(CK) levels,brain SOD activity and MDA and Ca 2+ levels were measured. Ultrastracture of brain tissue was examined by electron microscopy.Results Propofol 110 mg?kg -1 reduced mortality after brain ischemia/reperfusion injury. Infarction area of brain was significantly smaller in propofol and nimodipine groups than that in group 1. Propofol significantly inhibited the increases in serum LDH and CK levels induced by ischemia/reperfusion, increased SOD activity and decreased MDA content and Ca 2+ level in brain tissue. There was less brain tissue damage in propofol group.Conclusions Propofol 110 mg?kg -1 has protective effect against cerebral ischemia-reperfusion injury in rats.