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Role of C-Jun N-terminal kinase signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats

Na ZHANG ; Hongzhen LIU ; Xianjie WEN ; Yi LIU ; Xueqin ZHENG ; Chengxiang YANG

Chinese Journal of Anesthesiology.2013;33(9):1066-1069. doi:10.3760/cma.j.issn.0254-1416.2013.09.010

Objective To evaluate the role of C-Jun N-terminal kinase (JNK) signal transduction pathway in spinal neurotoxicity induced by lidocaine in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-260 g,were randomly divided into 6 groups (n =12 each):control group (group Ⅰ),sham operation group (group Ⅱ),JNK inhibitor group (group Ⅲ),dimethyl sulfoxide (DMSO) group (group Ⅳ),lidocaine group (group Ⅴ),and JNK inhibitor and lidocaine group (group Ⅵ).Group Ⅰ received no treatment.Intrathecal catheter was placed in the subarachnoid space in group Ⅱ.SP600125 25 μg and DMSO 20 μl were injected intrathecally in Ⅲ and Ⅳ groups,respectively.In group Ⅴ,10％ lidocaine 20 μl was intrathecally injected.SP600125 25 μg was injected intrathecally and 30 min later 10％ lidocaine 20 μl was injected intrathecally in group Ⅵ.Paw withdrawal threshold to yon Frey filament stimulation (PWT) and paw withdrawal latency to nociceptive thermal stimulation (PWL) were measured before intrathecal catheter was implanted (T0),before intrathecal administration (T1) and at 4,8 and 12 h and on 1,2,3,4,5 and 6 days after intrathecal administration (T2-10).At 24 h after intrathecal administration,4 rats were randomly chosen from each group and sacrificed.Their lumbar enlargements were removed for determination of phosphorylated JNK (p-JNK) expression (using Western blot) and neuronal apoptosis (by TUNEL).The apoptotic index was calculated.Results Compared with group Ⅰ,no significant difference was found in MWT and TWL in Ⅱ,Ⅲ groups and expression of p-JNK in Ⅱ and Ⅳ groups (P ＞ 0.05),MWT at T2-4,6-8 and TWL at T2-4,7 in group Ⅴ and MWT at T2-6 and TWL at T2-5 in group Ⅵ were significantly increased,the expression of p-JNK was down-regulated and the apoptotic index was decreased in group Ⅲ (P ＜ 0.05),and the expression of p-JNK was up-regulated and the apoptotic index was increased in Ⅴ and Ⅵ groups (P ＜ 0.05).Compared with group Ⅴ,MWT and TWL were significantly decreased,the expression of pJNK was down-regulated and the apoptotic index was decreased in group Ⅵ (P ＜ 0.05).Conclusion Activation of JNK signal transduction pathway is involved in spinal neurotoxicity induced by lidocaine in rats possibly through promoting neuronal apoptosis in the spinal cord.

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Effects of modified ultrafiltration on expression of aquaporin 1 in cardiopulmonary bypass-induced lung injury in dogs

Ming HAN ; Hong ZHANG ; Xuejiao DOU ; Huijun CAI ; Bangyong QIN ; Haiying WANG

Chinese Journal of Anesthesiology.2013;33(9):1070-1072. doi:10.3760/cma.j.issn.0254-1416.2013.09.011

Objective To evaluate the effects of modified ultrafiltration on the expression of aquaporin 1 (AQP1) in cardiopulmonary bypass (CPB)-induced lung injury in dogs.Methods Eighteen healthy adult dogs of either sex,weighing 15-20 kg,were randomly divided into 3 groups (n =6 each):control group (group C),group CPB and modified ultrafiltration group (group MUF).The dogs were anesthetized with intraperitoneal 2.5％pentobarbital 25 mg/kg.Thoracotomy was performed in all the three groups and in addition lung injury was produced by CPB in CPB and MUF groups.In group MUF,modified ultrafiltration was performed at 10-15 min after termination of CPB.Arterial blood samples were collected before mechanical ventilation (T1),at end of CPB (T2),and at 1 h after termination of CPB (T3) to calculate respiration index (RI) and oxygenation index (OI).The lungs were removed for microscopic examination of pathologic changes in lung tissues under light microscope and for detection of AQP1 mRNA expression by real-time PCR.Results RI and OI were significantly higher and AQP1 mRNA expression was lower at T2 and T3 than at T1 in CPB and MUF groups (P ＜ 0.05).Compared with group C,RI was significantly increased and AQP1 mRNA expression was down-regulated at T2,3 in CPB and MUF groups,and OI at T2.3 in CPB group and at T2 in MUF group was decreased (P ＜ 0.05).Compared with group CBP,RI was significantly decreased,OI was increased and AQP1 mRNA expression was up-regulated at T3 in group MUF (P ＜ 0.05).Conclusion Modified ultrafiltration can reduce CPB-induced lung injury in dogs and upregulation of AQP1 may be involved in the mechanism.

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Effects of dexmedetomidine on lidocaine-induced apoptosis in rat cortical neurons

Dongjian GE ; Bin QI ; Jinyu LI

Chinese Journal of Anesthesiology.2013;33(9):1079-1081. doi:10.3760/cma.j.issn.0254-1416.2013.09.014

Objective To evaluate the effects of dexmedetomidine on lidocaine-induced apoptosis in rat cortical neurons.Methods The cortical neurons obtained from Sprague-Dawley fetal rats were seeded in 24 multiwell plates at a density of 1 × 105 cells/ml,and the cortical neurons of 80 wells were randomly divided into 4 groups (n =20 each):control group (group C),lidocaine group (group L),dexmedetomidine group (group D) and lidocaine + dexmedetomidine group (group L+ D).The cells were cultured routinely in group C.The cells were exposed to lidocaine with a final concentration of 1 mmol/L in group L.The cells were exposed to dexmedetomidine with a final concentration of 3 μmol/L in group D.The cells were exposed to lidocaine and dexmedetomidine with the final concentrations of 1 mmol/L and 3 μmol/L,respectively,in group L + D.After 4 h incubation,the neurons were subjected to DAPI staining for detection of apoptosis,and the apoptosis rate was calculated.Western blot analysis was used to measure the expression of phosphorylated Akt (p-Akt),Akt and caspase-3.Results Compared with group C,the apoptosis rate was significantly increased,the expression of p-Akt was down-regulated,and the expression of caspase-3 was up-regulated in L and L + D groups (P ＜ 0.05),while no significant changes were found in the indexes mentioned above in group D (P ＞ 0.05).Compared with group L,the apoptosis rate was significantly decreased,the expression of p-Akt was up-regulated,and the expression of caspase-3 was down-regulated in L + D group (P ＜ 0.05).There was no significant difference in the expression of Akt between the four groups (P ＞ 0.05).Conclusion Dexmedetomidine can reduce lidocaine-induced apoptosis in rat cortical neurons,and activation of Akt may be involved in the underlying mechanism.

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Protective effect of perfluorocarbon emulsions on rat donor lung

Xuemei PENG ; Lu XI ; Huadong WANG ; Qingde ZHANG ; Huihui LIU ; Bing SHUAI

Chinese Journal of Anesthesiology.2013;33(9):1082-1084. doi:10.3760/cma.j.issn.0254-1416.2013.09.015

Objective To evaluate the protective effect of perfluorocarbon emulsions (FCE) on donor lung of rats during storage.Methods Twenty-four healthy male Sprague-Dawley rats,weighing 350-400 g,were equally and randomly divided into 2 groups using a random number table:University of Wisconsin (UW) solution group (UW group) and FCE group (FCE group).After the model of lung perfusion was established according to the method described by Fischer et al,the lung and heart were removed and perfused with 4 ℃ UW or FCE preservation solutions.The lung was taken out when stored for 6 h for determination of SOD activity (by WST assay),malondialdehyde (MDA) content (by TBA assay),and activity of myeloperoxidase (MPO) and content of interleukin1 β (IL-1β),IL-6,tumor necrosis factor-apha (TNF-α) (using ELISA) in lung tissues and for microscopic examination of pathologic changes.Results MPO activity was significantly lower in UW group than in FCE group (P ＜0.05).There were no significant differences in the SOD activity and content of MDA,IL-1β,IL-6,TNF-α between the two groups (P ＞ 0.05).Conclusion FCE can reduce the neutrophil infiltration in lung tissues,indicating that FCE is more superior to UW solution in reduction of injury to the donor lung of rats.

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Efficacy of different target concentrations of sufentanil TCI used to supplement topical anesthesia for fiber-optic bronchoscopy-assisted awake nasotracheal intubation in patients with obstructive sleep apnea syndrome

Xue XU ; Qiuyue DONG ; Xiang QI ; Wei LI ; Zhi LIANG ; Luchao GAO

Chinese Journal of Anesthesiology.2013;33(9):1089-1092. doi:10.3760/cma.j.issn.0254-1416.2013.09.017

Objective To compare the efficacy of different target concentrations of sufentanil target-controlled infusion used to supplement topical anesthesia for fiber-optic bronchoscopy (FOB)-assisted awake nasotracheal intubation in patients with obstructive sleep apnea syndrome (OSAS).Methods Forty-five ASA physical status Ⅱ or Ⅲ patients with OSAS,aged 28-60 yr,with body mass index of 30-40 kg/m2,scheduled for elective surgery,were randomly assigned into 3 groups (n =15 each):control group (group C) and sufentanil with the target plasma concentration of 0.4 ng/ml (group S1) and 0.6 ng/ml groups (group S2).Naso-pharyngeal and laryngeal mucous membrane was sprayed with 2％ lidocaine mixed with 1％ ephedrine for topical anesthesia in both groups.In addition 1％ tetracaine 3 ml was injected into trachea through cricothyroid membrane.FOB-assisted awake nasotracheal intubation was performed after the target concentration was achieved.The degree of airway obstruction was scored during intubation.The highest values of MAP and HR,rate-pressure product ＞ 12 000,decreased respiratory rate and hyoxemia were recorded during the period between induction of anesthesia and 3 min after intubation was completed.The changes in MAP and HR as percent of baseline values were calculated.Before topical anesthesia (T0),when target concentrations were reached (T1),and at 1 and 3 min after intubation (T2,3),blood samples were taken to determine the plasma concentrations of epinephrine (E),norepinephrine (NE) and cortisol.Results Compared with group C,the airway obstruction score was significantly decreased in group S1,the incidence of changes in MAP and HR ＞ 30％ of baseline values and rate-pressure product ＞ 12 000 was decreased,the plasma concentrations of E,NE and cortisol were decreased in S1 and S2 groups,and the incidence of the respiratory rate was decreased and hypoxemia was increased in group S2 (P ＜ 0.05).Compared with group S1,the airway obstruction score were significantly decreased,and the incidence of respiratory rate was decreased and hypoxemia was increased in group S2 (P ＜ 0.05).Compared with the baseline value at T0,the plasma concentrations of E,NE and cortisol were significantly increased at T2,3 in group C,while decreased at T1 in S1 and S2 groups (P ＜ 0.05).Conclusion Compared with pure topical anesthesia,sufentanil with the target plasma concentration of 0.4 ng/ml does not induce respiratory depression,maintains hemodynamics stable,attenuates the stress responses and provides better intubation conditions when used to supplement topical anesthesia for FOB-assisted awake nasotracheal intubation in patients with OSAS.

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Role of nitric oxide in spinal cord in maintenance of diabetic neuropathic pain in rats

Weicheng ZHAO ; Meijuan LIAO ; Chengxiang YANG ; Hanbing WANG ; Wanyou HE ; Hongzhen LIU ; Bin ZHANG

Chinese Journal of Anesthesiology.2013;33(9):1110-1113. doi:10.3760/cma.j.issn.0254-1416.2013.09.023

Objective To evaluate the role of nitric oxide (NO) in the spinal cord in the maintenance of diabetic neuropathic pain in rats.Methods Male Sprague-Dawley rats,aged 2 months,weighing 180-200 g,were used in the study.Diabetes mellitus was induced by intraperitoneal streptozotocin (STZ) 60 mg/kg and confirmed by blood glucose ＞ 16.7 mmol/L on day 2 after STZ injection.Twenty diabetic rats were randomly allocated into diabetes mellitus group (DM group,n =10) and L-NAME (non-selective NOS inhibitor) group (LN group,n =10).Another 10 age-matched normal rats served as control group (C group).On 21 days after STZ injection,L-NAME 10 mg/kg was injected intraperitoneally once a day for 7 consecutive days in LN group,whereas the equal volume of normal saline 5 ml/kg was given instead of L-NAME in DM group.Paw withdrawal threshold to yon Frey filament stimulation (PWT) was measured before STZ infection and on 7,14,21 and28 days after STZ injection.The rats were sacrificed after the last measurement of PWT and the lumbar segments of spinal cord were removed for determination of NO content and neuronal nitric oxide synthase (nNOS) expression (by Western blot analysis) in spinal cord tissues.Results Compared with C group,PWT was significantly decreased on 14,21 and 28 days after STZ injection,and the NO content and nNOS expression in spinal cord tissues were increased in DM and LN groups (P ＜ 0.05).Compared with DM group,PWT was significantly increased on 28 days after STZ injection,and the NO content and nNOS expression in spinal cord tissues were decreased in LN group (P ＜ 0.05).Conclusion NO in the spinal cord is involved in the maintenance of diabetic neuropathic pain in rats and the mechanism is related to the enhanced function of nNOS.

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Role of opioid receptors in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803

Li LI ; Dan HE ; Enjian GUAN ; Yuling ZHONG ; Yi QIN ; Yubo XIE

Chinese Journal of Anesthesiology.2013;33(9):1050-1052. doi:10.3760/cma.j.issn.0254-1416.2013.09.005

Objective To evaluate the role of opioid receptors in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was cultured in DMEM liquid culture medium.The cells were seeded in 6-well or 96-well plates and then randomly divided into 4 groups (n =54 each):control group (group C),fentanyl group (group F),naloxon group (group N) and naloxon + fentanyl group (group NF).The cells were exposed to 0.1 μmol/L fentanyl and 10 μmol/L naloxon in F and N groups,respectively.The cells were incubated with 10 μmnol/L naloxon for 30 min and then O.1 μmol/L fentanyl was added to the culture medium in group NF.The viability of the cells was detected by MTT assay after being incubated with fentanyl for 12,24,36,48,60 and 72 h.The cell apoptosis was assessed by flow cytometry after being incubated with fentanyl for 24 h.The migration of the cells was detected by wound healing assay after being incubated with fentanyl for 48 h.The proliferation of the cells was determined by colony formation assay at 7 day of incubation with fentanyl.Results Compared with group C,no significant changes in the viability of the cells,rate of colony formation,apoptotic rate and rate of cell wound healing were found in group N (P ＞ 0.05),and the viability of the cells,rate of colony formation and rate of cell wound healing were significantly decreased,and the apoptotic rate was increased in F and NF groups (P ＜ 0.05).There was no significant difference in the viability of the cells,rate of colony formation,rate of cell wound healing and apoptotic rate between group NF and group F (P ＞ 0.05).Conclusion Opioid receptors are not involved in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803 in vitro.

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Effect of long-term glucocorticoid administration on nondepolarizing muscle relaxants-induced neuromuscular blockade in rat diaphragma

Dan CHEN ; Lina HUANG ; Yuwei QIU ; Shitong LI

Chinese Journal of Anesthesiology.2013;33(9):1053-1055. doi:10.3760/cma.j.issn.0254-1416.2013.09.006

Objective To evaluate the effect of long-term glucocorticoid administration on nondepolarizing muscle relaxants-induced neuromuscular blockade in rat diaphragma in vitro.Methods Forty adult male SpragueDawley rats,aged 8 weeks,weighing 180-220 g,were randomized into 2 groups (n =20 each):control group (group C) and dexamethasone group (group D).In group D,dexamethasone 0.6 mg/kg was injected intraperitoneally once a day for 14 consecutive days,while the equal volume of normal saline was injected instead of dexamethasone in group C.The left phrenic nerve-hemidiaphragm was removed at 24 h after the last injection to evaluate the effect of d-tubocurarine.Different concentrations (0-10 μmol/L) of d-tubocurarine were added.The d-tubocurarine concentration-effect curve was drawn.The half inhibitory concentration and 95 ％ confidence interval of d-tubocurarine were calculated.Right diaphragma was removed for determination of the expression of embryonic nicotinic acetylcholine receptor (nAChRγ) mRNA and adult nicotinic acetylcholine receptor (nAChRε) mRNA by RTPCR.Results Compared with group C,the concentration-effect curve was shifted to the right,the half inhibitory concentration was significantly increased,and the expression of nAChRγ mRNA and nAChRε mRNA was up-regulated in group D (P ＜ 0.05).Conclusion Long-term glucocorticoid administration can weaken nondepolarizing muscle relaxants-induced neuromuscular blockade in rat diaphragma in vitro,and up-regulation of nAChR mRNA expression may be involved in the mechanism.

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Effects of pretreatment with nerve growth factor-beta on ischemia-reperfusion injury in isolated rat hearts:a comparison with preconditioning

Li AO ; Ke WEI ; Li LIU ; Su MIN

Chinese Journal of Anesthesiology.2013;33(9):1062-1065. doi:10.3760/cma.j.issn.0254-1416.2013.09.009

Objective To evaluate the effect of pretreatment with nerve growth factor-beta (NGF-β) on ischemia-reperfusion (I/R) injury in isolated rat hearts by comparing it with NGF-β preconditioning.Methods Pathogen-free male Sprague-Dawley rats,weighing 200-300 g,aged 8-10 weeks,were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.Their hearts were excised and perfused in a Langendorff apparatus with KH solution aerated with 95％ O2 and 5％ CO2 at 37℃.Twenty-four isolated rat hearts were randomly divided into 3 groups (n =8 each) using a random number table:I/R group,NGF-β pretreatment group (group NGFPt) and NGF-β preconditioning group (group NGFPc).The hearts were perfused with K-H solution for 10 min (equilibration) in each group.In group I/R,the hearts were continuously perfused with K-H solution for 30 min.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min before ischemia in group NGF-Pt.The hearts were continuously perfused with K-H solution containing NGF 0.1 μg/ml for 20 min followed by 10 min washout before ischemia in group NGFPc.The perfusion was suspended for 30 min followed by 120 min of reperfusion with K-H solution in each group.HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and + dp/dtmax were measured at the end of 10 min equilibration (baseline,T1),immediately before suspension of perfusion (T2),and at 5,30,60 and 120 min of reperfusion (T3-6).The activities of creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) in coronary effluent were measured at T1 and T3-6.Myocardial specimens were obtained at T6 for detection of myocardial apoptosis (by TUNEL) and for microscopic examination.Apoptotic index (AI) was calculated.Results Compared with the baseline value at T1,+ dp/dtmax was significantly decreased,and LVEDP and activities of CK-MB and LDH were increased at T3-6 in each group,LVDP and HR were decreased at T3-6 in group I/R,LVDP was decreased at T3,4 in group NGFPt and at T3-6 in group NGFPc,and HR was increased at T2-6 in NGFPt and NGF Pc groups (P ＜ 0.05).Compared with group I/R,LVDP,+ dp/dtmax and HR were significantly increased and LVEDP and activities of CK-MB and LDH and AI were decreased in NGFPt and NGFPc groups (P ＜ 0.05).Compared with group NGFPc,LVDP,+ dp/dtmax and HR were significantly increased,while the LDH activity and AI were decreased (P ＜ 0.05) and no significant changes were found in LVEDP and CK-MB activity in group NGFPt (P ＞ 0.05).The pathologic changes of myocardium were significantly reduced in NGFPt and NGFPc groups as compared with I/R group.Conclusion Pretreatment with 0.1 μg/ml NGF-β attenuates I/R injury in isloated rat hearts,and the efficacy is superior to that of NGF-β preconditioning.

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Effect of vagus nerve stimulation on blood coagulation and fibrinolysis in endotoxemic rats

Haisong WANG ; Zhaohui DU ; Li NIU ; Weiming CHEN ; Jianguo LI ; Yanlin WANG

Chinese Journal of Anesthesiology.2013;33(9):1121-1124. doi:10.3760/cma.j.issn.0254-1416.2013.09.026

Objective To evaluate the effect of vagus nerve stimulation on the blood coagulation and fibrinolysis in endotoxemic rats.Methods Ninety-six male Sprague-Dawley rats,weighing 250-280 g,were equally and randomly divided into 4 groups using a random number table:normal saline group (S group) ; lipopolysaccharide (LPS) group; vagus nerve cutting group (VNC group) and vagus nerve stimulation group (VNS group).Endotoxemia was induced by LPS 10 mg/kg injected via the left femoral vein.In group S,normal saline 5 ml/kg was injected via the left femoral vein.In S and LPS groups,the bilateral vagus nerves were only isolated.The bilateral vagus nerves were isolated,ligated and cut immediately after LPS injection in VNC group.The distal end of the vagus nerve was stimulated with direct current (5 V,2 ms,1 Hz) continuously for 20 min starting from the end of LPS injection in group VNS.Six rats were sacrificed before LPS injection (T0) and at 2,4 and 6 h after LPS injection (T1-3) and arterial blood samples were taken for determination of the levels of plasma tumor necrosis factorα (TNF-α),antithrombin (AT),tissue-type plasminogen activator (tPA),plasminogen activator inhibitor type 1 (PAI-1) and D-Dimer.Results Compared with group S,the plasma TNF-α,tPA,PAI-1 and D-Dimer levels were significantly increased,and AT level was decreased after LPS injection in LPS and VNC groups (P ＜ 0.05).Compared with group LPS group,the plasma AT level was significantly decreased,and the plasma PAI-1 level was increased in group VNC,and the plasma TNFα,tPA,PAI-1 and D-Dimer levels were decreased,and the plasma AT level was increased in VNS group (P ＜ 0.05).Conclusion Electrical stimulation of the vagus nerve can improve the blood coagulation and fibinonlysis in endotoxemic rats,and activation of cholinergic anti-inflammatory pathway,inhibition of inflammatory responses and reduction of damage to vascular endothelial cells may be involved in the mechanism.

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