Role of the autophagy in the treatment of neuropathic pain with pulsed radiofrequency / 中南大学学报(医学版)

Objective:To detect the effect of pulse radiofrequency (PRF) treatment on the neuropathic pain established by L5-spinal nerve ligation (SNL) on rats,and to investigate if PRF treatment would affect the expression of autophagy related protein LC3 and autophagy related receptor P62 at the dorsal horn.Methods:A total of 36 male Sprague-Dawley rats were randomly divided into 3 groups:a Sham group,a SNL group,and a SNL+PRF group.The 50％ paw withdrawal mechanical threshold (PWMT) was detected at 1 day before and 1,3,7,14 and 28 days post-operation by using Von-Frey filaments.The autophagy related protein LC3 and autophagy related receptor P62 were investigated by Western blot.Results:Compared with the Sham group,the PWMT significantly decreased in the SNL group at each time points (P＜0.05);in SNL+PRF group,PRF treatment could elevate the PWMT at the 1st day post-operation and lasted for 28 days (P＜0.05).What's more,SNL could elevate the LC3-Ⅱ and P62 levels at the 7th day post-operation (P＜0.05),which were decreased by the PRF treatment (P＜0.05).Conclusion:PRF treatment could improve SNL-induced the neuropathic pain,which might be partly due to the regulatory effects on the autophagy levels at the spinal dorsal horn.

Influence of ozonized saline on behaviors of rat models with incisional pain and levels of plasma TNF-α and IL-6 / 中国医师杂志

Objective To investigate the analgesia effect and the possible mechanism of intravenous administration with different concentrations of ozonized saline (OS) by observing changes in behavior,plasma tumor necrosis alpha (TNF-α),and interleukin-6 (IL-6) levels after rat plantar incision.Methods Fifty-four health adult male Sprague-Dawley rats were used in the investigation.Twenty four rats were randomly divided into four groups,6 in each group.The rats in group A1 were intravenously administered with 5ml/kg oxygen saline,10min before the incision.The rats in groups B1,C1,and D1 were intravenously administered with 20 μg/ml,30 μg/ml,and 40 μg/ml OS 5 ml/kg.An 1-cm incision was made in the right plantar surface from the heel to the toes according to the method described by Brennan under sevoflurane anesthesia.The 50％ paw withdrawal mechanical threshold (50％ PWMT) and cumulative pain score (CPS) at the times of 24 h before and 2,6,24,48 h after surgery were underwent.Thirty rats were randomly divided into five groups,6 in each group.Groups A2,B2,C2,and D2 processed as well as group 1.All groups except group E2 were made the model of incisional pain.2 ml blood was taken out of the right ventricular 2 h after the operation,then the levels of plasma TNF-α and IL-6 were detected by using enzyme linked immunosorbent assay (ELISA).Results There were no significant differences in the 50％PWMT between group A1 and group B1 at each time point after surgery (P ＞ 0.05).The 50％ PWMT in groups C1 and D1 were higher than those in group A1 at each time point after surgery (P ＜ 0.05).The CPS in groups B1,C1,and D1 were lower than those in group A1 after surgery (P ＜ 0.01).Compared to group E2,the levels of plasma TNF-α 2 h after the operation in group B2 and D2 were not statistically different (P ＞ 0.05).The levels of plasma TNF-α in groups C2 and A2 were higher than those in group E2 (P ＜ 0.05).The levels of plasma IL-6 2 h after the operation between group A2 and group E2 showed no difference (P ＞ 0.05).The levels of plasma IL-6 in groups B2,C2,and D2 were higher than those in group E2 (P ＜ 0.05).Concltsions Intravenous administration of ozonized saline can inhibit the incisional pain in rats.The analgesia effect of ozonized saline was dose-dependent.

Limb ischemic preconditioning reduces rabbit hepatic ischemia-reperfusion injury through inhibition the phosphorylation of proteins in the MAPK signal pathway in the late phase / 中南大学学报(医学版)

OBJECTIVE@#To investigate the liver protection mechanisms of MAPK signaling pathway of limb ischemia preconditioning in the late phase.@*METHODS@#Thirty-six adult male New Zealand white rabbits, weighing 1.8-2.0 kg, were randomly divided equally into 3 groups: group C (sham operation), group L (liver ischemia-reperfusion 24 h after limb ischemia preconditioning), group IR (liver ischemia-reperfusion without limb ischemia preconditioning). Serum alanine transaminase (ALT) was measured during ischemia reperfusion. The tissue and cell injury of liver were examined by optical and electron microscopy. Activation of P38MAPK, P44/P42MAPK, and JNK in hepatic tissue was assessed by western blot after 30 min of reperfusion.@*RESULTS@#Serum ALT and cell injury in the liver as examined by optical and electron microscopy was decreased in group L as compared with the group IR. Phosphorylation of P38MAPK, P44/ P42MAPK, and JNK were all increased significantly after 30 min of reperfusion. Phosphorylation of P38MAPK and JNK was reduced by limb ischemia pre-treatment.@*CONCLUSION@#Limb ischemia pre-treatment can induce the late phase of preconditioning in rabbit liver through the inhibition of the phosphorylation of P38MAPK and JNK.

Proteomics analysis of adenosine A1 receptor agonist-induced delayed myocardial protection in rabbits / 中南大学学报(医学版)

OBJECTIVE@#To investigate the changes of myocardial protein expression profiles in 2-chloro-N6-cyclopentyladenosine (CCPA), an adenosine A1 receptor agonist-induced delayed myocardial protection in New Zealand rabbits .@*METHODS@#A total of 8 rabbits were randomly divided into a CCPA group (CCPA group) and a normal saline group (NS group). CCPA and NS were infused into rabbits in the CCPA group and the NS group respectively. Twenty-four hours later, the rabbits were subjected to 30 min left anterior descending coronary artery occlusion and were reperfused for 2 hours, then the ischemic zone tissues of left ventricle were sampled for proteomic analysis.A total of 12 other New Zeland rabbits were divided into a sham group (Sham group), a normal saline group (NS group) and a CCPA group (CCPA group). The expression of αB-crystalline, one of the differential proteins, was confirmed by Western blot.@*RESULTS@#Analysis of two dimensional gel electrophoresis showed that the expression of 55 protein spots were different between the two groups, 17 protein spots were preliminarily identified with the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and Mascot and Expasy bioinformatics software. These proteins included stress proteins, metabolism-associated proteins, signal transduction pathway-related proteins, ionophorous proteins, immunity-associated proteins, and so on. Western blot showed that the expression of αB-crystalline was significantly up-regulated in the CCPA group.@*CONCLUSION@#The myocardial protein expression profiles are changed markedly in the preconditioning late phase of CCPA .The differential proteins might be involved in the delayed cardioprotection induced by CCPA.

Expression of bone morphogenetic proteins and their receptors in the normal adult rat spinal cord / 中南大学学报(医学版)

Objective To observe the expression distribution of bone morphogenetic proteins (BMP) in the spinal cord of normal adult rats. Methods Expression of BMP2, BMP4, and BMP7, and their receptors BMPR Ⅰa, BMPR Ⅰb, and BMP Ⅱ were detected by immunochemistry analysis in the spinal cord of normal adult rats. Results Expression of BMPR Ia or BMPR Ib was observed in the motor neurons of the anterior horn, sensory neurons of the dorsal horn, oligodentrocytes, some microglia, and some astrocytes. Expression of receptor BMPR Ⅱ was found in the oligodentrocytes and motor neurons in the gray matter of anterior horn. It was also expressed in some glial fibrillary acidic protein (GFAP)-positive astrocytes in the white matter but not in the gray matter. BMP2 and BMP4 were not expressed in the spinal cord of normal adult rats by immunohistochemistry. BMP7 was expressed in all the APC-positive oligodentrocytes, all the NeuN-positive motor neurons in the anterior horn, and some astrocytes in the normal spinal cord. Phosphated pSmad 1/5/8 protein was expressed in all the oligodentrocytes, all the neurons, and some astrocytes, especially in the GFAP-positive astrocytes which were RC2-positive radial glia in the subventricular zone.Conclusion BMP7, BMP receptors, and phosphated pSmad 1/5/8 are expressed in many types of cells whereas BMP2 and BMP4 are not expressed in the spinal cord of normal adult rats, which suggests an important function of BMP signal pathway in the neuron and glia of spinal cord.

Effect of high concentration carbon dioxide preconditioning on lipid peroxidation daring myocardial ischemia-reperfusion in rabbits / 中华麻醉学杂志

Objective To investigate the effect of high concentration carbon dioxide preconditioning on lipid peroxidation during myocardial ischemia-reperfusion (I/R) in rabbits. Methods Twenty-four New Zealand white rabbits weighing 2.0-3.9 kg were randomly divided into 3 groups ( n = 8 each): sham operation group (group S) , I/R group, high concentration carbon dioxide preconditioning group (group H) . The amimals were tracheal intubated and mechanically ventilated. In groups S and I/R, fresh gas flow was set at 0.3 L/min (100% O2 ), respiratory rate 30-40 bpm and tidal volume IS ml/kg, and PETCO2 was maintained at 40-50 mm Hg for 30 min. In group H, fresh gas flow was set at 0.3 L/min (100% O2), respiratory rate 20-30 bpm and tidal volume 10 ml/kg, PETO2 was maintained at 75-85 mm Hg for 5 min, and then all the ventilatory parameters were adjusted to the same as those in groups S and I/R. Myocardial I/R was produced by occlusion of left anterior descending branch of coronary artery for 30 min followed by 3 h reperfusion after preconditioning in groups I/R and H. The animals were sacrificed at the end of reperfusion and myocardial tissues obtained for determination of the superoxide dismutase (SOD) activity and malondialdehyde (MDA) content and examination of the ultrastnicture of myocardium with the transmission electron microscope. Results The SOD activity was significantly lower, while MDA content higher in group I/R than in group S ( P ＜ 0.01) . The SOD activity was significantly higher, while MDA content lower in group H than in group I/R ( P ＜ 0.01) . The myocardial injury was attenuated in group H compared with group I/R. ConclusionHigh concentration carbon dioxide preconditioning can reduce myocardial I/R injury in rabbits through inhibiting lipid peroxidation.

Proteomic analysis of morphine rabbit myocardium with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry / 中南大学学报(医学版)

OBJECTIVE@#To analyze the morphine rabbit myocardium with matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS).@*METHODS@#Six New Zealand white rabbits were randomly assigned to a control group (Group C) and a morphine group (Group M). Group C were pretreated with bolus injection of saline 1 mL/kg. Group M were pretreated with bolus injection of morphine 3 mg/kg. The myocardium tissue proteins of the rabbits 24 hours after the injection of morphine or saline preconditioned were extracted and separated by two dimensional gel electrophoresis(2-DE), and the images were analyzed and different proteins were found. Some of the different proteins were determined with MALDI-TOF-MS.@*RESULTS@#There were 51 protein spots that displayed quantitative changes in expression (P < 0.05), 15 protein spots were chosen for MS analysis, and 8 proteins were preliminarily identified.They were aldose reductase, zinc finger protein 312, src related tyrosine kinase, carbonic anhydrase 12 precursor, electron transfer flavoprotein beta-subunit, glyceraldehyde-3-phosphate dehydrogenase, tumor necrosis factor ligand superfamily member 11 and transmembrane emp24 domain-containing protein.@*CONCLUSION@#These proteins may be involved in the cardioprotection of morphine preconditioning.

KATP Channel Mediate the Protective Effect of Hypercarbonic Preconditioning on Myocardial Cells / 天津医药

Objective: To investigate the function of the ATP-sensitive K+(KATP) channel activation on the protective effect of hypercarbonic acidosis preconditioning on rabbit myocardial cells. Methods: Thirty-two rabbits were randomly divided into 4 groups (n = 8 for each group): pseudo-operation group (group P), ischemia and reperfusion group(group IR), hypercarbonic acidosis group(group H) and hypercarbonic acidosis+ glybenzcyclamide group (group H+G). Animals were ventilated normally in group IR and group P, tidal volume 15 mL/kg, breathing rate 35 bpm .The PETCO_2 was maintained at the level of 40-50 mm Hg for 30 minutes. Animals received low-frequency, low volume ventilation in group H group H+G, tidal volume 10 ml/kg, breathing rate 25 bpm to achieve hypercarbonic acidosis. The target value of PETCO_2 was 75-85 mm Hg. This value was maintained for 5 minutes. The animals then were ventilated normally to make the PETCO_2 return to 40-50 mm Hg. Animals were injected with 0.3 mg/kg glybenzcyclamide 10min before achieving hypercarbonic acidosis with hypoventilation in group H+G. Animals received ligation of left anterior branch artery for 30 minutes and reperfusion for 180 minutes in each group except P group. The myocardial ischemia area, the myocardial infarction area and their ratios were calculated by the ismaeil methods. Results: The ratio of the myocardial infarction area to the myocardial ischemia was significantly less in group H than those of group IR and group H+G (P 0.05). Conclusion: Hypercarbonic acidosis preconditioning can protect the cardiomyocytes by activating the KATP channel.

Delayed protective effect of morphine preconditioning on rabbit myocardium / 中南大学学报(医学版)

OBJECTIVE@#To investigate whether morphine preconditioning has the delayed protective effect on rabbit myocardium.@*METHODS@#Thirty New Zealand white rabbits were randomly divided into a NS group, a Mor-12 group and a Mor-24 group (n=10). In the Mor-12 group and Mor-24 group, morphine (3 mg/kg) was infused into rabbits, while the same volume of normal saline (NS) was given to rabbits in the NS group. Twelve hours after morphine infusion in the Mor-12 group, 24 h after NS or morphine infusion in the NS group and Mor-24 group, rabbits were subjected to 30 min left anterior descending coronary artery occlusions and were reperfused for 120 min. In 8 of the 10 rabbits in each group, arterial blood samples were taken before the ischemia (T1), 30 min after the ischemia (T2) and 120 min after the reperfusion (T3) to determine the concentration of cardiac troponin I (cTnI), and the myocardial infarct area was determined at the end of reperfusion. In the other 2 of the 10 rabbits in each group,the cell ultramicro-structure injury of myocardium was examined by electron microscope at the end of reperfusion.@*RESULTS@#The concentration of cTnI at T2 and T3 in the Mor-24 group was lower than that in the NS group and Mor-12 group.The myocardial infarct size, and cell ultramicrostructure injury of myocardium in the Mor-24 group were decreased compared with the NS group and Mor-12 group.@*CONCLUSION@#Morphine preconditioning has delayed protective effect on rabbit myocardium.

Effects of isoflurane delayed preconditioning on myocardial proteome in rabbits with myocardial ischemia-reperfusion injury / 中华麻醉学杂志

Objective To investigate the effects of isoflurane delayed preconditioning(IDP) on myocardial proteomin rabbits with myocardial ischemia-reperfuaion(I/R).mjury.Methods Eight New Zealand white rabbits of both sexes weighing 2.0-2.5 kg were randomly divided into 2 groups(n=4 each):I/R group and IDP group.Myocardial I/R Wgg induced by occlusion of left anterior descending artery for 40 min followed by 120 min repednsion.In group IDP.the animals inhaled 2%isoflurane for 2 h,undergoing I/R 24 h later.At the end of 120 min reperfusion,the myocardium of left ventricle anterior wall was removed for two-dimensional gel electrophorvsis.The different protein spots were analyzed by means of mass chromatography.Results There were 13 different protein spots between group I/R and IDP.Of the 13 proteins,the expression of 10 spots Was up-regulated and 3 spots down-regulated in quantity.Eleven protein spots of all spots were identified by means of MALDI-TOF-MS.Conclusion IDP can aNenuate myocardial I/R injury in rabbits and it may be related to the alteration in proteome of myocardium.

Effects of isoflurane delayed preconditioning on nuclear factor-κB during myocardial ischemia reperfusion injury in rabbits / 中华急诊医学杂志

Objective To explore the mechanism of the catdioprotection of isoflurane delayed preconditioning on myocardial ischemia reperfusion injury in rabbits.Method Thirty male New Zealand white rabbits,weighing 2.0 to 2.5 kg,were randomly divided into three groups(ten for each group):Control group(group C),I/R group(I/R group) ,2.0% isoflurane group(group S) .Group S was exposed to 2.0% isoflurane-100% oxygen for2 h.Group C and I/R group were exposed 2 h to 100% oxygen served as untreated controls.Twenty-four hours later I/R group and group S underwent 40 rain of coronary occlusion followed by 2 h of reperfusion.Blood samples were taken from arterial line at 20 min before occlusion(T1) ,20 rain after occlusion(T2) ,40 rain after occlusion(T3) ,1 h after reperfusion(T4) and 2 h after reperfusion(TS) for determination of plasma IL-10 levels and TNF-alevels by ELISA.At the end of the reperfusion,infarct size and area at risk were defined by Evans and TTC staining.The heart was harvested and levels of the nuclear factor kappa β(NF-κB)activity were determined by Western Blot,and ultrastructures were observed by electron microscopy.The data was expressed as,and were analyzed by using oneway ANOVA test with SPSS 13.0.P value less than 0.05 indicated statistical significance.Results The NF-κB activity of group S was significantly lower than that of group I/R(P<0.05).Group S significantly(P<0.05)reduced infarct size(19.7%±2.8% in group S) of the left ventricular area at risk as compared with control (37.8 %±1.7 % in I/R group).The injury of I/R group was worse than that of group S from the changes of the cellular structure under light microscope.Group S had a lower levels of TNF-α and also had a higher level of IL-10.Conclusions Isoflurane can inhibit NF-κB activity during myocardial ischemia reperfusion and modulate the cytokine expression,which may be one of molecular mechanisms of Isoflurane delayed preconditioning on cardioprotection.

Effects of propofol and fentanyl on myocardial ischemia-reperfusion injury in isolated rat heart / 中华麻醉学杂志

Objective To investigate the protective effect of propofol and/or fentanyl in myocardial ischemia-reperfusion injury. Methods Thirty-two SD rats weighing 250-300g were anesthetized with intraperitoneal ketamine 80 mg. Their hearts were excised and perfused with oxygenated (95%O2 and 5% CO2) K-H solution in a Langendorff apparatus at a perfusion pressure of 7.8 kPa. The temperature of perfusate and heart were kept at 37 ℃ . The experiment was divided randomly into four groups: A control group was perfused with K-H solution containing intralipid 90 ?g?kg-1 ( n = 8); B propofol group was perfused with K-H solution containing propofol 5?g?kg-1 ; C fentanyl group was perfused with K-H solution containing fentanyl 10 ng?kg-1; D propofol-fentanyl group was perfused with K-H solution containing propofol 5?g ?kg-1 and fentanyl 10 ng?kg-1. The isolated heart was perfused with above mentioned perfusate for 10 min followed by 50 min global normothermic (37℃) ischemia and 30 min reperfusion. The left ventricular developed pressure (LVDP) and ?dp/dtmax and coronary flow were measured 1 min before propofol and/or fentanyl perfusion, 1 min before global ischemia and at the end of 30 min reperfusion. The lactate dehydrogenase(LDH) activity in the total coronary effluent collected during the 30 min reperfusion was measured. Results The recovery of LVDP and ? dp/dtmax and coronary flow at the end of 30 min reperfusion were significantly better in group B, C and D than those in control group and the recovery was best in group D. The release of LDH in coronary effluent decreased significantly at the end of 30 min reperfusion in group B, C and D as compared with that in control group. LDH release was least in group D. Conclusions Both propofol and fentanyl can protect myocardium against ischemia- reperfusion injury in isolated rat hearts. The protective effect of propofol and fentanyl can be added and better protection can be provided.

Effects of diammonii glycyrrhizinatis on acute lung injury induced by cardiopulmonary bypass / 中华麻醉学杂志

Objective Diammonii glycyrrhizinatis was shown to exert a protective effect on myocardium against ischemia-reperfusion injury. The purpose of this study was to evaluate the effects of diammonii glycyrrhizinatis on acute lung injury induced by cardiopulmonary bypass(CPB), Methods Twenty-four ASA Ⅱ - Ⅲ patients (9 male, 15 female) aged 20-60 yr, weighing 45-75 kg scheduled for elective value replacement were divided into two groups at random: control group (group C, n = 12) and diammonii glycyrrhizinatis group (group G, n = 12) . Patients with infections diseases or immunodiflciency and those who were taking corticosteroid and drugs which may affect immune function were excluded. Anesthesia was induced and maintained with midazolam, fentanyl and vecuronium. In group G diammonii 2.5 mg?kg-1 10% glucose 50 ml was infused during the interval between intubation and skin incision. Blood samples were taken from arterial line at 10 min after induction (T0), 30 min after start of CPB (T1 ), 30 min after aortic unclamping (T2), at the end of surgery (T3), 4 h and 24 h after operation (T5, 6) for determination of plasma TNT-? IL-?, IL-10, MDA levels and Mn-SOD activity. Respiratory index (RI) PA-aDO2/PaO2 ] was calculated at 10 min after induction of anesthesia (T0), 10 min and 30 min after CPB (T1, 2) and at the end of surgery. Results There was no significant difference in age body weight, CPB time and aortic cross clamping time between the two groups. The TNF-?, IL-?, IL-10, MDA levels and Mn-SOD activity were not significanttly different at T0 between the two groups, and increased significanttly after start of CPB in both groups ( P