1.Effects of dexmedetomidine on lung ischemia-reperfusion injury in rats
Zhanhai WAN ; Hong ZHANG ; Yufang LENG ; Yongqiang LIU ; Xinghua LYU
Chinese Journal of Anesthesiology 2014;34(9):1066-1068
Objective To evaluate the effects of dexmedetomidine on lung ischemia-reperfusion (I/R) injury in rats.Methods Ninety-six healthy SPF male Wistar rats,weighing 250-350 g,aged 8-12 weeks,were randomly divided into 4 groups (n =24 each):sham operation group (S group),I/R group,low dose dexmedetomidine group (DL group) and high dose dexmedetomidine group (DH group).In DL and DH groups,dexmedetomidine 100 and 500 μg· kg-1 · d-1 were injected intraperitoneally once a day for 2 consecutive days,while the equal volume of normal saline was given in S and I/R groups.Lung·I/R was induced by clamping the left hilum of lung for 45 min followed by reperfusion at 30 min after administration on 2nd day.At 45 min of ischemia,and 60 and 120 min of reperfusion,6 rats were sacrificed,and lungswere removed for determination of TNF-α (tumor necrosis factor-a) content and myeloperoxidase (MPO) activity in lung tissues.The percentage of damaged alveolar in lung tissues was detected at 120 min of reperfusion.Another 6 rats were lavaged and the broncho-alveolar lavage fluid (BALF) was colleted for determination of the total protein concentrations.Results Compared with S group,the TNF-α content,MPO activity,percentage of damaged alveolar,and total protein concentrations in BALF were significantly increased in I/R,DL and DH groups.Compared with I/R group,the TNF-α content,MPO activity,percentage of damaged alveolar,and total protein concentrations in BALF were significantly decreased in DL and DH groups.Conclusion Dexmedetomidine can alleviate the lung I/R injury in rats,and the mechanism is related to inhibiton of the inflammatory responses.
2.Effect of dexmedetomidine on lipid peroxidation during lung ischemia-reperfusion in rats
Xinghua LYU ; Yufang LENG ; Yanni YANG ; Yuqing MA ; Xuanjie LI
Chinese Journal of Anesthesiology 2016;36(4):508-510
Objective To evaluate the effect of dexmedetomidine on lipid peroxidation during lung ischemia-reperfusion (I/R) in rats.Methods Fifty-four pathogen-free healthy male Wistar rats,weighing 250-350 g,were randomly divided into 3 groups (n=18 each) using a random number table:sham operation group (S group),I/R group,and dexmedetomidine group (Dex group).In group Dex,dexmedetomidine 5 μg/kg was injected via the caudal vein once a day for 2 consecutive days.The equal volume of normal saline was given in S and I/R groups.At 30 min after administration on 2nd day,lung I/R was produced by clamping the left hilum of lung for 45 min followed by 120 min of reperfusion in I/R and Dex groups.At 45 min of ischemia,and 60 and 120 min of reperfusion,6 rats selected from each group were sacrificed,and the left lungs were removed for examination of the pathological changes and for determination of wet/dry lung weight ratio (W/D ratio),malondialdehyde (MDA) content and superoxide dismutase (SOD) activity.Results Compared with S group,the SOD activity was significantly decreased,and the MDA content and W/D ratio were significantly increased at 60 and 120 min of reperfusion in I/R and Dex groups (P<0.05).Compared with I/R group,the SOD activity was significantly increased,and the MDA content and W/D ratio were significantly decreased at 60 and 120 min of reperfusion in Dex group (P<0.05).The pathological changes of lungs were significantly attenuated in Dex group as compared with I/R group.Conclusion Dexmedetomidine mitigates lung I/R injury through inhibiting lipid peroxidation in rats.
3.Screening and Structure Characterization of Acetylcholinesterase Inhibitors from Total Alkaloids of Fibraurea recisa Pierre.by Target Molecule Affinity-Liquid Chromatography-Tandem Mass Spectrometry
Zhongmei HE ; Na LYU ; Minlun NAN ; Yuwei ZHAO ; Yufang HE ; Lingwen MENG ; Jiaming SUN ; Lianxue ZHANG
Chinese Journal of Analytical Chemistry 2017;45(2):211-216
A target molecule affinity-ultrafiltration liquid chromatography-electrospray ionization tandem mass spectrometric (LC-ESI-MSn) method was established for rapid screening acetylcholinesterase (AChE) inhibitors from total alkaloids of fibraurea recisa Pierre.A total of 12 potential inhibitors were screened from Fibraurea recisa Pierre.and 6 compounds were identified including palmatine,berberine,jatrorrhizine,palmatrubine,7,8-dihydro-8-hydroxyberberine and groenlandicine.The AChE inhibitory activity of these 6 compounds was validated in vitro.Palmatine showed the strongest inhibitory activity for AChE,which was stronger than that of donepezil hydrochloride,demonstrating the potential of palmatine as anti-Alzheimer's drug.This method is simple,rapid,and accurate for directly screening active ingredients which can inhibit AChE from complex extract of traditional Chinese medicines.
4.Effects of sinomenine on expression of HIF-1α and VEGF during renal ischemia-reperfusion in rats
Peng WANG ; Yufang LENG ; Yujie SU ; Yanni YANG ; Guanzheng ZHENG ; Xinghua LYU
Chinese Journal of Anesthesiology 2015;35(8):1017-1019
Objective To evaluate the effects of sinomenine on the expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) during renal ischemia-reperfusion (I/ R) in rats.Methods Fifty-four male Wistar rats, weighing 180-220 g, were randomly divided into 3 groups (n=18 each) using a random number table: sham operation group (group S) , group I/R and sinomenine group (group SIN).Renal I/R was induced by clamping the left renal pedicle for 45 min followed by reperfusion in I/R and SIN groups.In group S, the bilateral renal pedicels were only exposed.Sinomenine 60 mg/kg was intraperitoneally injected at 30 min before reperfusion in group SIN, while the equal volume of normal saline was given in group S and group I/R.At 6, 12 and 24 h (T1-3) of reperfusion, 6 rats from each group were chosen, and blood samples were drawn from the hearts for determination of the serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.The animals were then sacrificed, and the left kidney was removed and embedded into a paraffin block for determination of the expression of HIF-1α and VEGF in the renal tissues (by immuno-histochemistry).Results Compared with group S, the serum Cr and BUN concentrations were significantly increased at T1-3 , and the expression of HIF-1α was significantly up-regulated at T2,3 in group I/R and group SIN, and the expression of VEGF was significantly upregulated at T2,3in group I/R, and at T1-3 in group SIN.Compared with group I/R, the serum Cr concentration at T1-3 and serum BUN concentration at T2,3 were significantly decreased and the expression of HIF-1α at T2,3and VEGF at T1-3was significantly up-regulated in group SIN.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to up-regulation of the expression of HIF-1α and VEGF in the renal tissues of rats.
5.Effect of sinomenine on levels of nuclear factor kappa B and inducible nitric oxide synthase during renal ischemia-reperfusion in rats
Yanni YANG ; Yufang LENG ; Guanzheng ZHENG ; Xinghua LYU ; Yujie SU ; Peng WANG
Chinese Journal of Anesthesiology 2015;35(8):951-954
Objective To investigate the effect of sinomenine on the levels of nuclear factor kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) during renal ischemia-reperfusion (I/R) in rats.Methods Seventy-two healthy male Wistar rats, aged 6-9 weeks, weighing 180-220 g, were randomly assigned into 4 groups (n =18 each) using a random number table: control group (group C), sinomenine group (group SIN), group I/R, and sinomenine+I/R group (group SIN+I/R).The animals were anesthetized with 10% chloral hydrate 350 mg/kg.The left renal pedicles were clamped with atraumatic microclips for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion to establish the model of renal I/R injury.Sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion in group SIN +I/R, and at the corresponding time point in group SIN.At 6, 8 and 12 h of reperfusion, the blood samples were drawn by cardiac puncture for measurement of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.The left renal specimens were obtained for examination of pathological changes (with light microscopes) and for determination of the rate of NF-κB-positive cells and iNOS expression in renal tissues (by immunohistochemistry).Results Compared with group C, the concentrations of serum Cr and BUN, and rate of NF-κB-positive cells were significantly increased, and the expression of iNOS was up-regulated in I/R and SIN+I/R groups, and no significant change was found in the parameters mentioned above in group SIN.Compared with group I/R, the concentrations of serum Cr and BUN, and rate of NF-κB-positive cells were significantly decreased, and the expression of iNOS was down-regulated in group SIN+I/R.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN+I/R compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activity of NF-κB and down-regulated expression of iNOS in rats.
6.Effect of sinomenine on apoptosis in renal tubular epithelial cells of rats subjected to renal ischemia-reperfusion: the relationship with JNK signaling pathway
Guanzheng ZHENG ; Yufang LENG ; Xinghua LYU ; Yujie SU ; Peng WANG ; Yanni YANG
Chinese Journal of Anesthesiology 2015;35(8):959-962
Objective To evaluate the effect of sinomenine on apoptosis in renal tubular epithelial cells of rats subjected to renal ischemia-reperfusion (I/R), and the relationship with C-Jun N-terminal kinase (JNK) signaling pathway.Methods Fifty-four male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were randomly divided into 3 groups (n =18 each) using a random number table: sham operation group (group S), I/R group and sinomenine group (group SIN).Renal ischemia was induced by occlusion of the left renal pedicle for 45 min followed by reperfusion, and the right kidney was removed immediately after onset of reperfusion in anesthetized rats in I/R and SIN groups.In group SIN, sinomenine 60 mg/kg was injected intraperitoneally at 30 min before reperfusion, while the equal volume of normal saline was given instead of sinomenine at the same time point in S and I/R groups.Six animals in each group were selected at 0.5, 6 and 24 h of reperfusion, blood samples were collected by cardiac puncture for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations.Immediately after blood sampling, the left kidney was removed for examination of pathological changes in renal tissues (with light microscopes) and for determination of phosphorylated JNK (p-JNK) and caspase-3 expression (by immune-histochemistry) and apoptosis in renal tubular epithelial cells (by TUNEL).The apoptotic rate was calculated.Results Compared with group S, the serum Cr and BUN concentrations were significantly increased, the expression of p-JNK and caspase-3 was up-regulated, and the apoptotic rate was increased in I/R and SIN groups.Compared with group I/R, the serum Cr and BUN concentrations were significantly decreased, the expression of p-JNK and caspase-3 was down-regulated, and the apoptotic rate was decreased in group SIN.The microscopic examination showed that the pathological changes of kidney were significantly attenuated in group SIN compared with group I/R.Conclusion The mechanism by which sinomenine attenuates renal I/R injury is related to inhibited activation of p-JNK signaling pathway and reduced apoptosis in renal tubular epithelial cells of rats.
7.Effect of lappaconitine on renal ischemia-reperfusion injury in mice
Yujie SU ; Yufang LENG ; Yanni YANG ; Peng WANG ; Guanzheng ZHENG ; Xinghua LYU
Chinese Journal of Anesthesiology 2015;35(4):510-512
Objective To evaluate the effect of lappaconitine on renal ischemia-reperfusion (I/R) injury in mice.Methods Thirty-six male Wistar rats,weighing 180-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and lappaconitine group (group LA).Renal I/R was induced by occlusion of the left renal pedicle for 45 min with atraumatic microclips followed by reperfusion,and the right kidney was removed after atraumatic microclips were released.At 30 min before reperfusion,lappaconitine 4 mg/kg was injected intraperitoneally in group LA,and normal saline 2 ml was given in S and I/R groups.In group S,the left renal pedicle was only isolated.At 5 and 24 h of reperfusion,blood samples were taken from the inferior vena cava for determination of serum creatinine (Cr) and blood urea nitrogen (BUN) concentrations,and kidney specimens were obtained for histopathologic examination (with light microscope) and for determination of the expression of cyclooxygenase-2 (COX-2) and matrix metalloproteinase-2 (MMP-2) in renal tissues (by immunohistochemistry).Results Compared with group S,the serum Cr and BUN concentrations were significantly increased,and the expression of COX-2 and MMP-2 in renal tissues was up-regulated at 5 and 24 h of reperfusion in I/R and LA groups.Compared with group I/R,the serum Cr and BUN concentrations were significantly decreased,the expression of COX-2 and MMP-2 in renal tissues was down-regulated at 5 and 24 h of reperfusion and histopathologic changes were reduced in group LA.Conclusion Lappaconitine can attenuate renal I/R injury through inhibiting the expression of COX-2 and MMP-2 in rats.
8.Protective effect of rhein lysinate on blood vessel damage induced by oxidative stress in mice and its mechanism
Qiusheng FENG ; Quan KAN ; Cuiping LYU ; Ran LI ; Jingbo WEI ; Yufang ZHAO ; Yongzhan ZHEN
Journal of Jilin University(Medicine Edition) 2015;(6):1171-1175
Objective To investigate the protective effects of rhein lysinate (RHL)on the blood vessel damage induced by oxidative stress in the mice,and to explore its mechanism.Methods The mouse models of oxidative damage were established by intraperitoneal injection of paraquat.30 C57 mice were randomly divided into control, paraquat model,and RHL prevention groups.The mice in RHL prevention group were given RHL by gavage for one week before performing model.The mice in other two groups were given equal volume of distilled water.For making model,paraquat was intraperitoneally injected in the mice in paraquat model and RHL prevention groups once a week for two weeks.The activities of superoxide dismutase (SOD)and glutathione peroxidase (GSH-Px) and the content of serum malonaldehyde (MDA) of the mice were detected 2 weeks after modeling. The pathological profile of blood vessel was observed by hematoxylin and eosin (HE)staining and the level of reactive oxygen species was observed by DCFH-DA staining.The expressions of genes related to blood vessel damage were detected by Western blotting method.Results Compared with control group,the activities of SOD and GSH-Px were decreased and the content of MDA was increased in paraquat model group (P < 0.05 ). Compared with paraquat model group,the activities of SOD and GSH-Px were increased and the content of MDA was decreased in RHL prevention group (P <0.05).The pathological examination indicated the structure of blood vessel of the mice was damaged and the level of reactive oxygen species of blood vessel was increased (P <0.05)in paraquat model group.The pathological changes were significantly improved and the level of reactive oxygen species of blood vessel of the mice was decreased (P < 0.05 )in RHL prevention group. The Western blotting analysis showed that compared with control group,the expression levels of nitric oxide endothelial synthase (eNOS)and caspase-3 of the mice in paraquat model group were decreased (P < 0.05),however the expression level of cleaved fragment of caspase-3 was increased (P < 0.05).Compared with paraquat model group,the expression levels of eNOS and caspase-3 of the mice in RHL prevention group were increased (P < 0.05 )and the expression level of cleaved fragment of caspase-3 was decreased (P <0.05).Conclusion Paraquat could induce vascular cell damage in vivo through increasing the levels of reactive oxygen species, and RHL could antagonize the effects of paraquat by scavenging reactive oxygen species, and up-regulating the eNOS expression and reducing the expression of the cleaved fragment of caspase-3.
9.Effect of intrathecal dexmedetomidine on expression of substance P and c-fos in spinal dorsal horns of rats with visceral pain
Guangru ZHANG ; Yufang LENG ; Xinghua LYU ; Chenmei PENG ; Fengxiang GU ; Jipeng LYU
Chinese Journal of Anesthesiology 2018;38(8):960-963
Objective To evaluate the effect of intrathecal dexmedetomidine on expression of sub-stance P (SP) and c-fos in the spinal dorsal horns of rats with visceral pain. Methods Thirty-two clean-grade healthy adult male Sprague-Dawley rats, weighing 250-300 g, were divided into 4 groups ( n=8 each) using a random number table method: control group (C group), visceral pain group (VP group), dexmedetomidine group (D group) and dexmedetomidine plus atipamezole group (DA group). VP, D and DA groups received intraperitoneal injection of 0. 9% acetic acid 10 ml∕kg to establish the model of visceral pain, while group C received the equal volume of normal saline instead. At 10 min before the model was es-tablished, dexmedetomidine 20 μl (1μg∕kg) and dexmedetomidine 1μg∕kg plus atipamezole 1μg∕kg (20μl) were intrathecally injected in D and DA groups, respectively, and the equal volume of normal saline 20μl was given instead in C and VP groups. Visceral pain index ( VPI) was recorded at 1 h after intraperito-neal injection, and then rats were sacrificed, and the dorsal horns of the spinal cord ( L4-6 ) were removed for determination of the expression of SP and c-fos by immunohistochemistry. Results Compared with group C, VPI was significantly increased, and the expression of SP and c-fos was up-regulated in VP, D and DA groups (P<0. 05). Compared with group VP, VPI was significantly decreased, and the expression of SP and c-fos was down-regulated in D and DA groups (P<0. 05). Compared with group D, VPI was signifi-cantly increased, and the expression of SP and c-fos was up-regulated in group DA ( P<0. 05) . Conclusion Intrathecal dexmedetomidine reduces the visceral pain through inhibiting the expression of SP and c-fos in the spinal dorsal horns, which is related to the α2-adrenergic receptor in spinal dorsal horns of rats.
10.Effect of dexmedetomidine on visceral pain in rats: the role of a2 adrenergic receptors in locus coeruleus
Chenmei PENG ; Yufang LENG ; Guangru ZHANG ; Fengxiang GU ; Jipeng LYU
Chinese Journal of Anesthesiology 2018;38(10):1227-1229
Objective To evaluate the effect of dexmedetomidine on visceral pain in rats and the role of α2 adrenergic receptors in locus coeruleus (LC).Methods Thirty-two healthy adult male SpragueDawley rats,weighing 250-300 g,were divided into 4 groups (n=8 each) using a random number table method:control group (group C),visceral pain group (group VP),dexmedetomidine group (group DEX) and α2-adrenergic receptor antagonist atipamezole group (group AP).α2-adrenergic receptor antagonist atipamezole 522 μg/kg was intramuscularly injected in group AP,and the equal volume of normal saline was given instead in C,VP and DEX groups.At 10 min after intramuscular injection,dexmedetomidine 10 μg/kg was injected via the tail vein in DEX and AP groups,and the equal volume of normal saline was given instead in C and VP groups.VP,DEX and AP groups received intraperitoneal injection of 0.9% acetic acid 10 ml/kg to make the visceral pain model at 15 min after injection via the tail vein.The cumulative visceral pain score was recorded within 60 min after acetic acid injection.The number of c-fos positive cells in LC was detected by immunohistochemistry,and the content of norepinephrine (NA) in the spinal cord were detected by enzyme-linked immunosorbent assay at 2 h after acetic acid injection.Results Compared with group C,the cumulative visceral pain scores,the number of c-fos positive cells in LC and content of NA in the spinal cord were significantly increased in VP,DEX and AP groups (P<0.05).Compared with group VP,the cumulative visceral pain scores,the number of c-fos positive cells in LC and content of NA in the spinal cord were significantly decreased in DEX and AP groups (P<0.05).Compared with group DEX,the cumulative visceral pain scores,the number of c-fos positive cells in LC and content of NA in the spinal cord were significantly increased in group AP (P<0.05).Conclusion Dexmedetomidine can alleviate visceral pain in rats,and the mechanism is partially related to activating α2 adrenergic receptors in LC.