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1.
Article in Chinese | WPRIM | ID: wpr-911295

ABSTRACT

Objective:To evaluate the role of p38 mitogen-activated protein kinase (MAPK)/cyclic adenosine monophosphate response element-binding protein (CREB) signaling pathway in tetramethylpyrazine-induced reduction of hippocampal inflammatory responses in mice with sepsis-associated encephalopathy (SAE).Methods:Sixty healthy male C57BL6 mice, weighing 24-27 g, were divided into 4 groups ( n=15 each) using a random number table method: sham operation group (group Sham), sepsis group (group Sep), tetramethylpyrazine group (group TMP) and p38 MAPK inhibitor SB203580 group (group SB). The model of SAE was established by cecal ligation and puncture in anesthetized mice.Tetramethylpyrazine 10 mg/kg was injected intraperitoneally once a day at 3 days before the establishment of the model in TMP group, and SB203580 2.0 mg/kg was intraperitoneally injected at 30 min after the establishment of the model in SB group.The equal volume of normal saline was given intraperitoneally in Sham and Sep groups.At 1 day after operation, cognitive function was assessed by Morris water maze, and the escape latency and ratio of time spent in the target quadrant were recorded.The animals were sacrificed after the test, and hippocampal tissues were taken for determination of the contents of interleukin-1beta (IL-1β), tumor necrosis factor-alpha (TNF-α) and IL-6 (by enzyme-linked immunosorbent assay) and for detection of the expression of phosphorylation of p38 MAPK, GSK3 and CREB and expression of brain-derived neurotrophic factor (BDNF) (by Western blot). Results:Compared with group Sham, the escape latency was significantly prolonged, the ratios of time spent in the target quadrant were decreased, the contents of IL-1β, TNF-α and IL-6 were increased, the phosphorylation of hippocampus p38 MAPK was increased, the phosphorylation of GSK3 and CREB were decreased, and the expression of BDNF was down-regulated in Sep, TMP and SB groups ( P<0.05). Compared with group Sep, the escape latency was significantly shortened, the ratios of time spent in the target quadrant were increased, the contents of IL-1β, TNF-α and IL-6 were decreased, the phosphorylation of hippocampus p38 MAPK was decreased, the phosphorylation of GSK3 and CREB were increased, and the expression of BDNF was up-regulated in TMP and SB groups ( P<0.05). Compared with group TMP, no significant change was found in the parameters mentioned above in group SB ( P>0.05). Conclusion:p38 MAPK/CREB signaling pathway is involved in the process of tetramethylpyrazine-induced reduction of hippocampal inflammatory responses in mice with SAE.

2.
Article in Chinese | WPRIM | ID: wpr-911187

ABSTRACT

Objective:To evaluate the role of extracellular signal-regulated kinase 1/2 (ERK1/2)/cyclic adenosine monophosphate response element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway in dexmedetomidine-induced inhibition of propofol-caused apoptosis in isolated hippocampal neurons of fetal rats.Methods:Pregnant Sprague-Dawley rats at 16 days of gestation were sacrificed, and the fetal rats were taken out, and hippocampal neurons of fetal rats were obtained and primarily cultured in vitro for 7 days.The neurons were divided into 9 groups ( n=12 each) using a random number table method: control group (group C), fat emulsion group (group I), dimethyl sulfoxide (DMSO) group, dexmedetomidine group (group D), propofol group (group P), propofol plus dexmedetomidine group (group PD), PD98059 plus propofol plus dexmedetomidine group (group PDP), MH89 plus propofol plus dexmedetomidine group (group HDP), and KG501 plus propofol plus dexmedetomidine group (group KDP). Group C received no treatment.In group I, 20% fat emulsion was added, and the neurons were incubated for 30 min, and 0.25% DMSO was added in group DMSO, and the neurons were incubated for 30 min.Dexmedetomidine at a final concentration of 10 μmol/L was added, and the neurons were incubated for 30 min in group D. Propofol at a final concentration of 100 μmol/L was added, and the neurons were incubated for 3 h in group P. In group PD, dexmedetomidine at a final concentration of 10 μmol/L was added, the neurons were incubated for 30 min, propofol at a final concentration of 100 μmol/L was added, and the neurons were incubated for 3 h. In PDP, HDP and KDP groups, 25 μmol PD98059 (p-ERK1/2 inhibitor), 10 μmol H89 (p-CREB inhibitor) and 25 μmol KG501 (CREB inhibitor) were added, respectively, the neurons were incubated for 30 min, dexmedetomidine at a final concentration of 10 μmol/L was added, the neurons were incubated for 30 min, and propofol at a final concentration of 100 μmol/L was added, and the neurons were incubated for 3 h. The cell ultrastructure was observed with the transmission electron microscope, the apoptosis in neurons was detected by flow cytometry, the expression of ERK1/2, CREB and BDNF mRNA was detected by quantitative real-time polymerase chain reaction, and the expression of p-ERK1/2, CREB, p-CREB, BDNF and cleaved caspase-3 was detected by Western blot. Results:Compared with group C, the apoptosis rate was significantly increased, the expression of p-ERK1/2 and p-CREB was down-regulated, and the expression of cleaved caspase-3 was up-regulated in P, PD, PDP, HDP and KDP groups, and the expression of BDNF was significantly down-regulated in P, PDP, HDP and KDP groups ( P<0.05). Compared with group P, the apoptosis rate was significantly decreased, the expression of p-ERK1/2, p-CREB and BDNF was up-regulated, and the expression of cleaved caspase-3 was down-regulated in group PD ( P<0.05). Compared with group PD, the apoptosis rate was significantly increased, the expression of p-ERK1/2, p-CREB and BDNF was down-regulated, and the expression of cleaved caspase-3 was up-regulated in PDP, HDP and KDP groups ( P<0.05). Conclusion:The ERK1/2/CREB/BDNF signaling pathway is involved in dexmedetomidine-induced inhibition of propofol-caused apoptosis in isolated hippocampal neurons of fetal rats.

3.
Acta Pharmaceutica Sinica B ; (6): 734-745, 2020.
Article in English | WPRIM | ID: wpr-828846

ABSTRACT

Peroxisome proliferator-activated receptor (PPAR) is a transcriptional coactivator that binds to a diverse range of transcription factors. PPAR coactivator 1 (PGC-1) coactivators possess an extensive range of biological effects in different tissues, and play a key part in the regulation of the oxidative metabolism, consequently modulating the production of reactive oxygen species, autophagy, and mitochondrial biogenesis. Owing to these findings, a large body of studies, aiming to establish the role of PGC-1 in the neuromuscular system, has shown that PGC-1 could be a promising target for therapies targeting neuromuscular diseases. Among these, some evidence has shown that various signaling pathways linked to PGC-1 are deregulated in muscular dystrophy, leading to a reduced capacity for mitochondrial oxidative phosphorylation and increased reactive oxygen species (ROS) production. In the light of these results, any intervention aimed at activating PGC-1 could contribute towards ameliorating the progression of muscular dystrophies. PGC-1 is influenced by different patho-physiological/pharmacological stimuli. Natural products have been reported to display modulatory effects on PPAR activation with fewer side effects in comparison to synthetic drugs. Taken together, this review summarizes the current knowledge on Duchenne muscular dystrophy, focusing on the potential effects of natural compounds, acting as regulators of PGC-1.

4.
Article in English | WPRIM | ID: wpr-759498

ABSTRACT

BACKGROUND: The pain-relief properties of tricyclic antidepressants can be attributed to several actions. Recent observations suggest that adenosine is involved in the antinociceptive effect of amitriptyline. The A3 adenosine receptor (A3AR) is the only adenosine subtype overexpressed in inflammatory and cancer cells. This study was performed to investigate the role of A3AR in the anti-nociceptive effect of amitriptyline. METHODS: Spinal nerve-ligated neuropathic pain was induced by ligating the L5 and L6 spinal nerves of male Sprague-Dawley rats. The neuropathic rats were randomly assigned to one of the following three groups (8 per group): a neuropathic pain with normal saline group, a neuropathic pain with amitriptyline group, and a neuropathic pain with amitriptyline and 3-ethyl-5-benzyl- 2-methyl-4-phenylethynyl-6-phenyl-1,4-(±)-dihydropyridine-3,5-dicarboxylate (MRS) group. Amitriptyline or saline was administered intraperitoneally and 3-ethyl-5-benzyl-2-methyl-4-phenylethynyl-6-phenyl-1,4-(±)-dihydropyridine-3,5-dicarboxylate (MRS-1191), an A3AR antagonist, was injected subcutaneously immediately before amitriptyline administration. The level of extracellular signal-regulated kinase P44/42 (ERK1/2), cyclic AMP response element-binding protein (CREB), and proinflammatory cytokines were assessed using immunoblotting or reverse-transciption polymerase chain reaction. RESULTS: Amitriptyline increased the mechanical withdrawal threshold of the neuropathic rats. The level of phospho-ERK1/2 and phospho-CREB proteins, and proinflammatory cytokines produced by spinal nerve ligation were significantly reduced by amitriptyline administration. However, the use of MRS-1191 before amitriptyline administration not only reduced the threshold of mechanical allodynia, but also increased the signaling protein and proinflammatory cytokine levels, which were reduced by amitriptyline. CONCLUSIONS: The results of this study suggest that the anti-nociceptive effect of amitriptyline involves the suppression of ERK1/2 and CREB signaling proteins, and A3AR activation also affects the alleviation of the inflammatory response.


Subject(s)
Adenosine , Amitriptyline , Animals , Antidepressive Agents, Tricyclic , Cyclic AMP Response Element-Binding Protein , Cytokines , Humans , Hyperalgesia , Immunoblotting , Ligation , Male , Neuralgia , Phosphotransferases , Polymerase Chain Reaction , Rats , Rats, Sprague-Dawley , Receptors, Purinergic P1 , Spinal Nerves
5.
Chinese Journal of Anesthesiology ; (12): 1288-1292, 2018.
Article in Chinese | WPRIM | ID: wpr-745591

ABSTRACT

Objective To evaluate the role of cyclic adenosine monophosphate-protein kinase A-cAMP response element-binding protein (cAMP-PKA-CREB) signaling pathway in hypoxic preconditioninginduced reduction of propofol-induced central neurotoxicity in the developing rats.Methods A total of 70 SPF male Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 7 groups (n=10 each) using a random number table method:normal saline group (N group),propofol group (P group),hypoxic preconditioning plus propofol group (HP group),hypoxic preconditioning plus propofol plus PKA inhibitor H89 group (HPH group),propofol plus PKA agonist SP-CAMP group (PS group),normal saline injected via the lateral cerebral ventricle group (NI group),and 5% dimethyl sulfoxide (DMSO) injected via the lateral cerebral ventricle group (DI group).In P group,propofol 50 mg/kg was intraperitoneally injected,and an increment of propofol 50 mg/kg was given after recovery of righting reflex.The equal volume of normal saline was given instead in N group.In HP group,hypoxic preconditioning (rats were subjected to 5 cycles of 10-min hypoxia of 8% O2 and 1O-min normoxia of 21% O2) was performed,and propofol was intraperitoneally injected at 2 h after the end of hypoxic preconditioning and the method was similar to those previously described in P group.In HPH group,H89 5 μmol/5 μl was injected via the lateral cerebral ventricle,and 30 min later the other treatment was similar to those previously described in HP group.In PS group,SP-CAMP 20 nmol/5 μl was injected via the lateral cerebral ventricle,and 30 min later propofol was injected using the method previously described in P group.In NI and DI groups,5 μl normal saline and 5% DMSO were injected via the lateral cerebral ventricle,respectively.Rats were immediately sacrificed after the righting reflex was recovered,brains were removed and hippocampi were isolated and cut into sections which were stained with haematoxylin and eosin for determination of PKAc and p-CREB positive cells (by i mmuno-histochemistry) and expression of cleaved caspase-3,Bcl-2,Bax,PKAc and posphorylated (p-CREB) protein (by Western blot).Results Compared with N group,the expression of cleaved caspase-3 and Bax was significantly up-regulated,the expression of Bcl-2,PKAc and p-CREB was downregulated,and the percentage of PKAc and p-CREB positive cells was decreased (P<0.05),hippocampal cells had irregular arrangement,and cells was atrophied in P group.Compared with P group,the expression of cleaved caspase-3 was significantly down-regulated,the expression of Bcl-2,PKAc and p-CREB was up-regulated,and the percentage of PKAc and p-CREB positive cells was increased in HP and PS groups,and the expression of Bax was down-regulated (P<0.05),the hippocampal cells were arranged neatly,the cytoplasm was abundant,and the nuclei were visible in HP group.Compared with HP group,the expression of cleaved caspase-3 was significantly up-regulated,the expression of Bcl-2,PKAc and p-CREB protein was down-regulated and the percentage of PKAc and p-CREB positive cells was decreased (P<0.05),the cells had irregular arrangement and shrinked,and nuclear condensation was found in cells in HPH group.Conclusion The mechanism by which hypoxic preconditioning reduces propofol-induced central neurotoxicity may be related to activating cAMP-PKA-CREB signaling pathway in the developing rats.

6.
Chinese Journal of Anesthesiology ; (12): 1177-1180, 2018.
Article in Chinese | WPRIM | ID: wpr-734648

ABSTRACT

Objective To evaluate the effect of sevoflurane on hippocampal cyclic AMP (cAMP)-protein kinase A (PKA)-cAMP response element-binding protein (CREB) signaling pathway in aged rats.Methods Sixty healthy aged male Sprague-Dawley rats,aged 18 months,weighing 600-750 g,were divided into control group (C group,n =30) and sevoflurane group (group Sev,n =30) using a random number table method.Group Sev inhaled 2% sevoflurane for 4 h.Group C inhaled the mixture of 50% air and oxygen (2 L/min) for 4 h.Morris water maze test was performed on 1-6 days before anesthesia and at 1 day after the end of anesthesia (at the corresponding time points in group C).The animals were sacrificed and hippocampi were removed at 1,3 and 7 days after anesthesia (at the corresponding time points in group C) for determination of the hippocampal cAMP content (using enzyme-linked immunosorbent assay) and expression of hippocampal CREB,phosphorylated CREB (p-CREB) and PKA (using Western blot),and the p-CREB/CREB ratio was calculated.Results Compared with group C,the escape latency and swimming distance were significantly prolonged,the frequency of crossing the original platform was decreased,the time of staying at the original platform quadrant was shortened at 1 day after the end of anesthesia,and the expression of hippocampal cAMP and PKA was down-regulated at 1,3 and 7 days after the end of anesthesia,and the expression of CREB and p-CREB was down-regulated and p-CREB/CREB ratio was decreased at 1 day after the end of anesthesia in group Sev (P< 0.05).Conclusion The mechanism of sevoflurane-induced postoperative cognitive dysfunction may be related to inhibiting hippocampal cAMP-PKA-CREB signaling pathway in aged rats.

7.
Article in Chinese | WPRIM | ID: wpr-608222

ABSTRACT

Objective To evaluate the relationship between hippocampal cyclic adenosine monophosphate response element-binding protein(CREB)/brain-derived neurotrophic factor(BDNF)signaling pathway and cognitive dysfunction in rats with chronic pathological pain.Methods Thirty-two healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups using a random number table:control group(group C,n=8),sham operation group(group S,n=8)and chronic pathological pain group(group CP,n=16).Chronic pathological pain model was established by injecting cobra venom 0.4 mg(4 μl)into the sheath of the infraorbital nerve.The mechanical pain threshold was measured at 3 days before establishment of the model(baseline)and 4 days and 1,2,3,4 and 8 weeks after establishment of the model.Morris water maze test was performed to evaluate the spatial learning and memory abilities at 5 and 9 weeks after establishment of the model.Eight rats were sacrificed at 5 and 9 weeks after establishment of the model in CP group,and rats were sacrificed after the end of Morris water maze test at 9 weeks after establishment of the model in C and S groups.The hippocampi were isolated for determination of the expression of phosphorylated CREB and BDNF in the hippocampal tissues using Western blot.Results Compared with group C,the mechanical pain threshold was significantly decreased at each time point after establishment of the model,the escape latency was prolonged at 5 and 9 weeks after establishment of the model,the rate of time of staying at the target quadrant was decreased,the frequency of crossing the original platform was decreased,and the expression of phosphorylated CREB and BDNF was down-regulated at 9 weeks after establishment of the model in group CP(P0.05).Conclusion The mechanism underlying cognitive dysfunction may be related to inhibited activation of CREB/BDNF signaling pathway in the hippocampus of rats with chronic pathological pain.

8.
Article in Chinese | WPRIM | ID: wpr-513927

ABSTRACT

Objective To evaluate the effect of sevoflurane on hippocampal calcium/calmodulindependent protein kinase Ⅱ (CaMK Ⅱ)/cyclic adenosine monophosphate response element-binding protein (CREB) signaling pathway in aged rats.Methods Sixty pathogen-free healthy male Sprague-Dawley rats,aged 18 months,weighing 600-750 g,were divided into 2 groups (n=30 each) using a random number table:control group (group C) and sevoflurane group (group Sev).Group Sev inhaled 2% sevoflurane in the mixture of 50% air and oxygen (2 L/min) for 4 h.Group C inhaled the mixture of 50% air and oxygen (2 L/min) for 4 h.Morris water maze test was performed on 6 days before anesthesia and 1 day after anesthesia.The escape latency,swimming distance,frequency of crossing the original platform and time of staying at the platform quadrant Ⅱ were recorded.On 1,3 and 7 days after anesthesia,the rats were sacrificed,and the hippocampus was obtained for determination of the expression of CaMK Ⅱ,phosphorylated CaMK Ⅱ,CREB and phosphorylated CREB by Western blot.Results Compared with group C,the escape latency and swimming distance were significantly prolonged,the frequency of crossing the original platform was decreased,and the time of staying at the platform quadrant Ⅱ was reduced on 5th day of training and 1 day after anesthesia,and the expression of CaMK Ⅱ,phosphorylated CaMK Ⅱ,CREB and phosphorylated CREB was down-regulated after anesthesia in group Sev (P< 0.05).Conclusion Sevoflurane leads to cognitive decline through inhibiting hippocampal CaMK Ⅱ/CREB signaling pathway in aged rats.

9.
Article in Chinese | WPRIM | ID: wpr-924124

ABSTRACT

@#Objective To explore the effects of electroacupuncture at Shenting (DU24) and Baihui (DU20) on cognitive dysfunction after stroke. Methods Forty-five Sprague-Dawley rats were randomly divided into control group (n=15), model group (n=15) and electroacupuncture group (n=15). The latter two groups were occluded their middle cerebral artery for two hours and reperfused. The electroacupuncture group accepted electroacupuncture at Shenting and Baihui 24 hours after modeling for seven days. They were assessed with Morris water maze once a day since the second day of intervention. Their brains were stained with TTC staining to measure cerebral infarction volume after treatment, while the expression of cyclic AMP response element binding protein (CREB) and phosphorylation (p-CREB) in hippocampal CA1 area were detected with immunohistochemistry. Results The escape latency and swimming distance of place navigation shortened in the electroacupuncture group compared with those in the model group (P<0.05) from the fourth day of intervention. The number of cross platform of spatial probe increased (P<0.05). The infarction volume was less in the electroacupuncture group than in the model group (P< 0.05), with increased expression of CREB and p-CREB in hippocampal CA1 area (P<0.05). Conclusion Electroacupuncture at Shenting and Baihui can increase the expression of CREB and phosphorylation in hippocampal CA1 area in rats after cerebral ischemia-reperfusion, to protect the neurons from ischemia and improve the learning and memory function.

10.
Article in English | WPRIM | ID: wpr-80631

ABSTRACT

PURPOSE: Pelvic irradiation for the treatment of cancer can affect normal cells, such as the rapidly proliferating spermatogenic cells of the testis, leading to infertility, a common post-irradiation problem. The present study investigated the radioprotective effect of rolipram, a specific phosphodiesterase type-IV inhibitor known to increase the expression and phosphorylation of the cyclic adenosine monophosphate response element-binding protein (CREB), a key factor for spermatogenesis, with the testicular system against pelvic irradiation. MATERIALS AND METHODS: Male C57BL/6 mice were treated with pelvic irradiation (2 Gy) and rolipram, alone or in combination, and were sacrificed at 12 hours and 35 days after irradiation. RESULTS: Rolipram protected germ cells from radiation-induced apoptosis at 12 hours after irradiation and significantly increased testis weight compared with irradiation controls at 35 days. Rolipram also ameliorated radiation-induced testicular morphological changes, such as changes in seminiferous tubular diameter and epithelial height. Additionally, seminiferous tubule repopulation and stem cell survival indices were higher in the rolipram-treated group than in the radiation group. Moreover, rolipram treatment counteracted the radiation-mediated decrease in the sperm count and mobility in the epididymis. CONCLUSIONS: These protective effects of rolipram treatment prior to irradiation may be mediated by the increase in pCREB levels at 12 hours post-irradiation and the attenuated decrease in pCREB levels in the testis at 35 days post-irradiation in the rolipram-treated group. These findings suggest that activation of CREB signaling by rolipram treatment ameliorates the detrimental effects of acute irradiation on testicular dysfunction and the related male reproductive functions in mice.


Subject(s)
Adenosine Monophosphate , Animals , Apoptosis , Cyclic AMP Response Element-Binding Protein , Epididymis , Germ Cells , Humans , Infertility , Male , Mice , Phosphorylation , Rolipram , Seminiferous Tubules , Sperm Count , Spermatogenesis , Stem Cells , Testis
11.
Article in Chinese | WPRIM | ID: wpr-482098

ABSTRACT

OBJECTIVE To investigate the effect of injection of β2-adrenergic receptor agonist clenbuterol into the infralimbic cortex(IL) on drug-seeking behavior triggered by conditioned cues. METHODS Adult male SD rats were trained to self-administer heroin under a FR1 schedule for consecutive 14 d,followed by 2-h extinction training. Cue-induced heroin seeking was measured for 2 h. Clenbuterol was microinjected bilaterally into the IL(8 ng/side)of rats 15 min prior to reinstatement test. Meanwhile,locomotor activity was detected 15 min after clenbuterol or artifial cerebrospinal fluid(mod?el group) was microinjected bilaterally into IL. Western blotting was used to detect the expression of phosphorylated cyclic AMP response element-binding protein(p-CREB)in the prelimbic cortex(PL), IL,nucleus accumbens core (NACc) and shell (NACsh) of rats immediately after reinstatement test. RESULTS After heroin administration training for 14 consecutive days,these animals exhibited reliable heroin self-administration,indicated by the increase in active nose poke responses and infusions. The rats that had received infusion of clenbuterol into the IL had significantly lower active pokes (8 ± 3)than those in model group(45±10)in cue-induced reinstatement(P<0.01),but there was no significant differ?ence between clenbuterol group and vehicle group in the locomotor activity. The expression of p-CREB in either IL or NACsh was significantly decreased in clenbuterol group compared with model group(P<0.01,P<0.05),but significantly increased in NACc(P<0.01). CONCLUSION Microinjection of clenb?uterol into the IL can attenuate the cue-induced reinstatement of heroin-seeking behavior in rats. The underlying mechanism might be related to the regulation of p-CREB expression in the NACc and NACsh.

12.
Article in Chinese | WPRIM | ID: wpr-479901

ABSTRACT

Objective To investigate the relationship between spinal neuronal microRNA 212 (miR-212) and phosphorylation of cAMP response element-binding protein (CREB) in a mouse model of bone cancer pain (BCP).Methods Thirty-two male SPF C3H/HeJ mice, aged 4-6 weeks, weighing 20-25 g, were randomly divided into 4 groups (n=8 each) using a random number table: sham operation group (group S), BCP group, BCP + intrathecal negative control locked nucleic acid (LNA) group (group BC) , and BCP + intrathecal miR-212 antisense LNA group (group BL).After the mice were anesthetized with intraperitoneal pentobarbital sodium, 20 μl of α minimal essential medium containing NCTC 2472 cells 2×105 was injected directly into the medullary cavity of the distal femur.In BC and BL groups, negative control LNA and miR-212 antisense LNA 12 pmol/5 μl were intrathecally injected, respectively, once a day for 7 consecutive days, starting from day 14 after inoculation.In S and BCP groups, the equal volume of DNAse/RNAse-free water was given instead.The number of spontaneous flinches (NSF) and mechanical paw withdrawal threshold (MWT) were measured on 1 day before inoculation and 4, 7, 10, 14 and 21 days after inoculation.The mice of each group were sacrificed after measurement of pain threshold on 21 days after inoculation, and the lumbar enlargement segments of the spinal cord were harvested to detect the expression of phosphorylated CREB (p-CREB) and CREB using Western blot.Results Compared with group S, the MWT was significantly decreased, and the NSF was increased on 7-21 days after inoculation, and the expression of p-CREB was up-regulated in BCP, BC and BL groups.Compared with group BCP, the MWT was significantly increased, and the NSF was decreased on 21 days after inoculation, and the expression of p-CREB was down-regulated in group BL, and no significant change was found in the parameters mentioned above in BC group.There was no significant difference in the expression of CREB between the four groups.Conclusion Spinal neuronal miR-212 is involved in the maintenance of BCP probably by promoting phosphorylation of CREB in mice.

13.
Article in Chinese | WPRIM | ID: wpr-479879

ABSTRACT

Objective To evaluate the effect of sevoflurane anesthesia on the expression of phosphorylated cAMP response element-binding protein (p-CREB) in the hippocampal neurons of developing rats.Methods Thirty-two healthy male Sprague-Dawley rats, aged 7 days, weighing 10-15 g, were equally and randomly divided into either control group (group C) or sevoflurane anesthesia group (group Sev) using a random number table.Group C inhaled 30% oxygen for 6 h.Group Sev inhaled 3% sevoflurane for 6 h.Eight rats in each group were sacrificed immediately after the end of oxygen or sevoflurane inhalation, and the hippocampus was removed for determination of the expression of p-CREB.The rats at ages 2 months underwent Morris water maze test.The rats were then sacrificed, and the hippocampus was removed for determination of the expression of p-CREB by Western blot.Results Compared with group C, the escape latency and swimming distance were significantly prolonged, the frequency of crossing the original platform was decreased, the percentage of the time of staying at the quadrant Ⅱ was decreased, and the expression of p-CREB in hippocampal neurons was down-regulated in group Sev.Conclusion The mechanism of sevoflurane anesthesia-induced neurotoxicity is related to inhibition of p-CREB expression in hippocampal neurons of developing rats.

14.
Journal of Medical Postgraduates ; (12): 149-152, 2015.
Article in Chinese | WPRIM | ID: wpr-461174

ABSTRACT

Objective Beta platelet-derived growth factor receptor ( PDGFR-β)-mediated signaling plays a key role in mor-phine tolerance , but its molecular mechanisms are not yet completely understood .The present study aims to investigate whether the ex-tracellular signal-regulated kinase ( ERK) and cyclic AMP response element binding protein ( REB) signaling pathways are involved in the development of PDGFR-βactivation-induced morphine tolerance in rats . Methods Thirty-six adult male SD rats were randomly divided into six groups of equal number:normal saline (20μL), morphine (15μg), morphine +imatinib (morphine 15μg +ima-tinib 10μg), morphine +PDGF-BB (morphine 15μg +PDGF-BB 10 ng), imatinib (10μg), and PDGF-BB (10 ng), all treated intrathecally at 20μL once daily for 7 consecutive days .Paw withdrawal latency ( PWL ) was measured 1 d before and 30 min after medication at 1, 3, 5, and 7 days, respectively, followed by calculation of the maximal possible effect of analgesia (MPE).On the 8th day, PWL was again obtained from all the rats at 30 min after intrathecal injection of morphine (15μg).Then, all the animals were sacrificed and the L4-5 segment of the spinal cord was isolated for determination of the expressions of ERK , phosphorylated ERK ( p-ERK) , CREB, and phosphorylated CREB ( p-CREB) by Western blot. Results At 5 and 7 days after medication, MPE was significant decreased in the morphine group ([52.90 ±8.20] and [15.12 ±3.80] %) and the morphine +PDGF-BB group ([43.51 ±5.42] and [14.81 ±3.60] %) as compared with (100.00 ± 0.00) %in both groups at 1 day (P<0.05), but had no significant changes in the morphine +imatinib group at 1, 3, 5, and 7 days.After intrathecal injection of morphine on the 8th day, MPE was (16.22 ±2.51) %in the morphine group, (15.22 ±3.50) %in the morphine +PDGF-BB group, and (35.21 ±4.51) %in the PDGF-BB group, all remarkably lower than (100.00 ±0.00) %in the control group (P<0.05).There were no significant differences in the expression levels of ERK and CREB among the six groups.The expressions of spinal p-ERK and p-CREB were markedly increased in the morphine , morphine +PDGF-BB, and PDGF-BB groups as compared with the control group (P<0.05), but significantly decreased in the morphine +imatinib group in compari-son with the morphine group, (P<0.05). Conclusion The PDGFR-βsignaling pathway plays an important role in the develop-ment of tolerance to morphine-induced analgesia and its underlying mechanisms may be associated with the activation of the ERK and CREB pathways .

15.
Chinese Journal of Anesthesiology ; (12): 1309-1311, 2014.
Article in Chinese | WPRIM | ID: wpr-468487

ABSTRACT

Objective To evaluate the effects of dexmedetomidine on the expression of phosphor-cAMP response element binding protein (p-CREB) in isoloated hippocampal neurons of fetal rats.Methods SpragueDawley rats on 16-18 days of gestation were sacrificed and the fetal rats were obtained.The hippocampi of fetal rats were isolated and hippocampal neurons were seeded in culture medium for 8 days.The cells were then divided into 4 groups (n =12 each) using a random number table:control group (group C),dexmedetomidine 0.001 μmol/L group (group D1),dexmedetomidine 0.010 μmol/L group (group D2),and dexmedetomidine 0.100μmol/L group (group D3).In D1.3 groups,dexmedetomidine with the final concentrations of 0.001 μmol/L,0.010 μmol/L,and 0.100 μmol/L was added to the culture medium,respectively,and then the cells were incubated for 3.5 h.The apoptosis in hippocampal neurons was detected by flow cytometry.The expression of p-CREB in hippocampal neurons was determined by RT-PCR and Western blot.Results Compared with group C,apoptosis rate was significantly decreased and the expression of p-CREB was up-regulated in D1.3 groups.Conclusion Dexmedetomidine inhibits apoptosis in isolated hippocampal neurons of fetal rats by up-regulating the expression of p-CREB.

16.
Journal of Medical Postgraduates ; (12): 686-689, 2014.
Article in Chinese | WPRIM | ID: wpr-453324

ABSTRACT

Objective Research has indicated that hydrogen sulfide(H2S) can regulate the function of N-methyl-D-aspartate re-ceptors(NMDARs) in the brain, but its effect on brain resuscitation requires further investigation.The study was to speculate the effect of H2 S on brain resuscitation as well as the underlying mechanism of neuroresuscitation by investigating the effects of hydrogen sulfide and hypo-thermia on the expression of NR2A, NR2B and phospho-cAMP response element binding protein (p-CREB) of NMDARs in the hippocampus after global cerebral ischemia following by reperfusion. Methods 100 male SD rats were randomly divided into five groups(n=20):sham operation group, model group, mild hypothermia group, NaHS group, NaHS combined mild hypothermia group.Pulsinelli-Brierley four-ves-sel occlusion method was induced to build the injury rat model by reperfusion after global cerebral ischemia .After 15 minutes'ischemia, im-mediate injection of 14μmol/kg NaHS was performed intraperitoneally on NaHS group and NaHS combined mild hypothermia group , while skin cooling(rectal temperature=32-33℃) was done on mild hypothermia group and NaHS combined mild hypothermia group .6 hours late,r hip-pocampus were extracted from rat heads.Respectively, spectrophotometer was applied to measure the content of H2S, Western blot for the expres-sions of NR2 A,NR2 B and pC-REB, and RTP-CR for mRNA level of brain derived neurotrophic (BDNF). HE staining was also performed on brain tissues 72hours after reperfusion on 4 rats from each group to evaluate the pathological changes of pyramidal neurons in CA1 region. R esul ts The content of H 2 S increased in each of the four groups after ischemia-reperfusion compared with sham operation group ( 15.2 ±2.0 nmol/g) (P0.05).The gray values of NR2A and NR2B in each group increased compared with sham operation group(P1 in NaHS group and NaHS combined mild hy-pothermia group.Compared with the expression of p-CREB(0.55 ±0.06) in model group, there were significant increases in mild hypother-mia group(0.99 ±0.15), NaHS group(1.05 ±0.12), NaHS combined mild hypothermia group(1.02 ±0.15)(P<0.05).Compared with the expression of BNDF mRNA(0.83 ±0.12) in model group, there were significant increases in mild hypothermia group (1.11 ±0.13), NaHS group(1.27 ±0.16), NaHS combined mild hypothermia group(1.35 ±0.16)(P<0.05).In comparison to model group, there were signifi-cant alleviation in the injury of pyramidal neurons in hippocampal CA1 region in mild hypothermia group, NaHS group, NaHS combined mild hypothermia group, with the best effect in NaHS combined mild hypothermia group . Conclusion Hydrogen sulfide combined mild hypo-thermia can selectively activate synaptic NMDA receptors and trigger the prosurvival CREB signaling pathway to exert brain resuscitation .

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Article in Chinese | WPRIM | ID: wpr-446811

ABSTRACT

Objective To evaluate the effects of dexmedetomidine on the activity of cAMP response element binding protein (CREB) and c-fos in the spinal dorsal horn in a rat model of neuropathic pain.Methods Fifty-four adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =18 each):sham operation group (group S),chronic neuropathic pain group (group C) and dexmedetomidine group (group D).The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.The sciatic nerve was exposed and 4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread in C and D groups.In group D,dexmedetomidine 50 μg/kg was injected intraperitoneally once a day starting from the end of operation until 1 day before the animals were sacrificed,while the equal volme of normal saline was injected instead of dexmedetomidine in S and C groups.Paw withdrawal threshold to mechanical stimulation with yon Frey filament (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured on 1 day before operation and 3,7 and 14 days after operation.The animals were sacrificed after measurement of MWT and TWL.Their lumbar segments (L4-6) of the spinal cord were removed for measurement of the expression of phosphorylated CREB (pCREB) and c-fos by immunohistochemistry.Results Compared with group S,MWT was significantly decreased,TWL was shortened,and the expression of pCREB and c-fos was up-regulated on 3,7 and 14 days after operation in C and D groups (P < 0.05).Compared with group C,MWT was significantly increased,TWL was prolonged,and the expression of pCREB and c-fos was down-regulated on 3,7 and 14 days after operation in group D (P < 0.05).MWT was significantly lower,and TWL was shorter on 3,7 and 14 days after operation than on 1 day before operation in C and D groups (P < 0.05).MWT was significantly lower,TWL was shorter,and the expression of pCREB and c-fos was higher on 7 and 14 days after operation than on 3 days after operation in C and D groups (P < 0.05).MWT was significantly higher,TWL was longer,and the expression of pCREB and c-fos was lower on 14 days after operation than on 7 days after operation in C and D groups (P < 0.05).Conclusion The mechanism by which dexmedetomidine reduces neuropathic pain is related to inhibition of the activity of CREB and c-fos in the spinal dorsal horn of rats.

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Article in Chinese | WPRIM | ID: wpr-442846

ABSTRACT

Objective To evaluate the effects of curcumin on the expression of phosphorylated extracellular signal-related kinase (p-ERK) and phosphorylated cAMP response element binding protein (p-CREB) in the spinal dorsal horn and dorsal root ganglion (DRG) in type 2 diabetic neuropathic pain (DNP) in rats.Methods Type 2 diabetes mellitus was induced by high-fat and high-sucrose diet and intraperitoneal streptozotocin (STZ) 35mg/kg,and confirmed by fasting blood glucose level≥ 16.7 mmol/L in male Sprague-Dawley rats.Type 2 DNP was confirmed by the mechanical withdrawal threshold (MWT) and thermal withdraw latency (TWL) measured on day 14 after STZ administration < 80% of the baseline value,and the rats with type 2 DNP were randomly divided into 3 groups (n =27 each):type 2 DNP group (group DNP),curcumin group (group Cur) and solvent control group (group SC).Curcumin and corn oil 100 mg/kg (25 mg/ml) were injected intraperitoneally once a day for 14consecutive days starting from 14 days after administration of streptozocin in Cur and SC groups,respectively.Another 27 normal rats were served as control group (group C) and were fed with common forage.MWT and TWL were measured at 3,7 and 14 days after curcumin injection (T1 3),and the lumbar segment 4-6 of the spinal cord and DRGs were removed at the same time for determination of the expression of p-ERK and p-CREB (by Western blot).Results Compared with group C,MWT was significantly decreased,TWL was shortened,and the expression of p-ERK and p-CREB in spinal dorsal horn and DRGs was up-regulated at T1-3 in DNP and SC groups,and at T1 in Cur group (P < 0.05).Compared with group DNP,MWT was significantly increased,TWL was prolonged,and the expression of p-ERK and p-CREB in spinal dorsal horn and DRGs was down-regulated at T2,3 in Cur group (P <0.05).There was no significant difference in the MWT,TWL and expression of p-ERK and p-CREB between DNP and SC groups (P > 0.05).Conclusion Curcumin can attenuate type 2 diabetic DNP by inhibiting up-regulation of the expression of p-ERK and p-CREB in the spinal dorsal horn and DRG in rats.

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Chinese Journal of Anesthesiology ; (12): 1198-1201, 2013.
Article in Chinese | WPRIM | ID: wpr-438965

ABSTRACT

Objective To evaluate the effects of propofol anesthesia on hippocampal protein kinase A (PKA)-cAMP response element binding protein (CREB) signaling pathway in neonatal rats.Methods One hundred and seventy-five male Sprague-Dawley rats,aged 7 days,weighing 8-15 g,were randomly divided into 5 groups (n =35 each) using a random number table:control group (C group) and propofol 25,50,100 and 200 mg/kg groups (P~ groups).Groups P1 and P2 received intraperitoneal propofol 25 and 50 mg/kg,respectively.Groups P3 and P4 received intraperitoneal propofol 100 and 200 mg/kg,respectively,and after righting reflex completely recovered,an increment of propofol 50 mg/kg was given until the total amount was finished.Five animals in each group were chosen and arterial blood samples were obtained immediately after the animals were fully awake for blood gas analysis.Five animals in each group were chosen at 2 h after fully awake and the age of 9 weeks,the rats were sacrificed and their brains were removed for microscopic examination of the ultrastructure of hippocampal neurons and for determination of PKA mRNA,CREB mRNA,PKA protein and pCREB protein in hippocampus (using RT-PCR and Western blot analysis).Results There was no significant difference in the indexes of blood gas analysis anong the five groups (P > 0.05).Nuclear swelling and fragmentation,chromatin condensation,apoptotic bodies,decreased number of synapses and widened synaptic space were observed in P2,P3 and P4 groups.Compared with group C,the expression of PKA mRNA,CREB mRNA,PKA protein and pCREB protein was significantly down-regulated at 2 h after fully awake and the age of 9 weeks in P1,P2,P3 and P4 groups (P < 0.05).Conclusion The mechanism by which propofol anesthesia induces neurotoxicity in neonatal rats may be related to inhibition of the activity of PKA-CREB signaling pathway.

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Article in Chinese | WPRIM | ID: wpr-418903

ABSTRACT

Objective To evaluate the role of extracelluar signal-regulated kinase (ERK)-cyclic AMP response element binding protein (CREB) signaling pathway in the spinal cord in naloxone-induced withdrawal response in morphine-dependent rats.Methods Fifty male adult SD rats,aged 2 months,weighing 200-250 g,in which intrathecal catheters were successfully implanted without complications,were randomly divided into 5 groups (n =10 each):group control (group C); group morphine dependence (group MD); group morphine withdrawal (group MW); group U0126 (ERK signaling pathway blocker); group dimethyl sulfoxide (DMSO,solvent for U0126).Morphine dependence was induced by increasing doses of subcutaneous morphine for 6 days.The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg twice every other day until 50 mg/kg on 6th day in groups MD,MW,U0126 and DMSO.Morphine withdrawal response was induced by intraperitoneal naloxone 4 mg/kg at 4 h after last morphine administration in groups MW,U0126 and DMSO.U0126 150μg (in DMSO 10 μl) and DMSO 10 μl were administered intrathecally at 30 min before naloxone administration in groups U0126 and DMSO respectively.Morphine withdrawal response (0=no withdrawal response,3 =severe response)and touch evoked agitation (0 =no agitation,2 =severe agitation) were observed and scored during 1 h after naloxone administration.The animals were then sacrificed and the spinal cord was removed for determination of the expression of phosphorylated ERK (p-ERK) and phosphorylated CREB (p-CREB) by immuno-histochemistry and Western blot.Results Morphine withdrawal significantly up-regulated the p-ERK and p-CREB expression in group MW compared with group C ( P < 0.05).Withdrawal response score and touch evoked agitation score were significantly increased in groups MW,U0126 and DMSO as compared with group MD ( P < 0.05).U0126 pretreatment significantly attenuated naloxone-induced increase in withdrawal response score and touch evoked agitation score and down-regulated p-ERK and p-CREB expression in group U0126 as compared with group MW ( P < 0.05).Conclusion ERK-CREB signaling pathway in the spinal cord is involved in morphine withdrawal response in morphine-dependent rats.

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