Effect of activating adenosine A2B receptors on autophagy during myocardial ischemia-reperfusion in rats: role of PI3K/Akt signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of activating adenosine A2B receptors on autophagy during myocardial ischemia-reperfusion (I/R) and the role of phosphatidylinositol 3-kinase/serine/threonine protein kinase (PI3K/Akt) signaling pathway in rats.Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, weighing 220-280 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (group Sham), myocardial I/R group (group I/R), adenosine A2B receptor agonist BAY 60-6583 group (group BAY) and BAY 60-6583+ PI3K inhibitor LY 294002 group (group BAY+ LY). Myocardial I/R was induced by occlusion of the anterior descending branch of the left coronary artery for 30 min followed by 120-min reperfusion.BAY 60-6583 1 mg/kg was intraperitoneally injected at 5 min before reperfusion in group BAY.BAY 60-6583 1 mg/kg was intraperitoneally injected at 5 min before reperfusion and LY 294002 10 mg/kg was intraperitoneally injected at 10 min before reperfusion in group BAY+ LY.Blood samples were obtained at the end of reperfusion for determination of concentrations of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB) in serum (by enzyme-linked immunosorbent assay). The animals were sacrificed, and myocardial tissues were obtained for measurement of the percentage of myocardial infarct size (by Evan Blue and TTC double-staining) and for determination of the expression of microtubule-associated protein 1 light chain 3 (LC3Ⅰ), LC3Ⅱ, Beclin-1 and phosphorylated Akt (p-Akt) (by Western blot). The ratio of LC3Ⅱ/LC3Ⅰ was calculated. Results:Compared with group Sham, the serum LDH and CK-MB concentrations and percentage of myocardial infarct size were significantly increased, the expression of p-Akt was down-regulated, the expression of Beclin-1 and LC3Ⅱ was up-regulated, and the ratio of LC3Ⅱ/LC3Ⅰ was increased in group I/R ( P<0.05). Compared with group I/R, the concentrations of serum LDH, CK-MB and percentage of myocardial infarct size were significantly decreased, the expression of p-Akt was up-regulated, the expression of Beclin-1 and LC3Ⅱ was down-regulated, and the ratio of LC3Ⅱ/LC3Ⅰ was decreased in the group BAY ( P<0.05), and no significant change was found in the parameters mentioned above in group BAY+ LY ( P>0.05). Compared with group BAY, the concentrations of serum LDH, CK-MB and percentage of myocardial infarct size were significantly increased, the expression of p-Akt was down-regulated, the expression of Beclin-1 and LC3Ⅱ was up-regulated and the ratio of LC3Ⅱ/LC3Ⅰ was increased in group BAY+ LY ( P<0.05). Conclusion:Activating adenosine A2B receptors can decrease autophagy of myocardial cells during myocardial I/R injury, and the mechanism may be related to activating PI3K/Akt signaling pathway in rats.

Mechanism of mental dependence of propofol in rats: adenosine A2A receptor-neurotransmitter-ERK pathway / 中华麻醉学杂志

Objective:To investigate the relationship between the mechanism of mental dependence of propofol and adenosine A2A receptor-neurotransmitter-extracellular signal-regulated kinase (ERK) pathway in rats.Methods:Forty-eight healthy male Sprague-Dawley rats, aged about 7 weeks, weighing 200-300 g, were used in this study.The model of propofol dependence was established by intraperitoneal injection of propofol 40 mg/kg for 14 consecutive days.The rats were divided into 6 groups ( n=8 each) using a random number table method: central control group (group c-C), central agonist group (group c-CGS), central antagonist group (group c-DMPX), peripheral control group (group p-C), peripheral agonist group (group p-CGS) and peripheral antagonist group (group p-DMPX). Adenosine A2A agonist CGS-21680 2.5 ng/0.5 μl was intracranially injected immediately after establishing the model in group c-CGS, while the equal volume of normal saline was given instead in c-C group.CGS-21680 0.1 mg/kg was intraperitoneally injected in group p-CGS, while the equal volume of normal saline was given instead in group p-C.Adenosine A2A receptor antagonist DMPX 50 ng/0.5 μl was intracranially injected at 20 min before each propofol injection in group c-DMPX, and DMPX 0.25 mg/kg was intraperitoneally injected in group p-DMPX.The position preference value (CPP value) was determined before establishing the model, immediately after establishing the model, and after administration of agonist or normal saline (after intervention). The animals were sacrificed at 1 day after establishing the model, and blood samples and brain tissues were obtained for determination of the levels of dopamine (DA) and glutamate (Glu) in plasma and hippocampus and content of serotonin (5-HT) in cerebral cortex (by enzyme-linked immunosorbent assay) and expression of phosphorylated ERK1/2 (p-ERK1/2) in cerebral cortex (by Western blot). Results:Compared with the baseline before establishing the model, CPP value was increased immediately after establishing the model in c-C, c-CGS, p-C and p-CGS groups ( P<0.05), and no significant change was found in CPP value immediately after establishing the model in c-DMPX and p-DMPX groups ( P>0.05). Compared with the value immediately after establishing the model, no significant change was found in CPP value after intervention in c-C and p-C groups ( P>0.05), and CPP value was increased after intervention in c-CGS and p-CGS groups ( P<0.05). Compared with group c-C, the contents of hippocampal DA and Glu were significantly increased in group c-CGS, and the contents of hippocampal Glu were decreased, the content of 5-HT in cerebral cortex was increased, and the expression of p-ERK1/2 was down-regulated in group c-DMPX ( P<0.05). Compared with group p-C, no significant change was found in levels of DA and glutamate (Glu) in plasma and hippocampus and 5-HT and p-ERK1/2 in cerebral cortex in group p-CGS ( P>0.05), and the contents of hippocampal DA and Glu were significantly decreased, the content of 5-HT in cerebral cortex was increased, and the expression of p-ERK1/2 was down-regulated in group p-DMPX ( P<0.05). Conclusion:The mechanism underlying the development of propofol mental dependence may be related to activating adenosine A2A receptors, increasing excitatory neurotransmitters in brain, and thus up-regulating ERK activity in rats.

Inhibition of the neutral sphingomyelinase-2 pathway protects against cerebral ischemia reperfusion injury in rats / 国际脑血管病杂志

Objective To investigate the roles of neutral sphingomyelinase-2 (nSMase2) pathway on cerebral edema and cerebral injury in cerebral ischemia-reperfusion injury in rats. Methods Seventy-six adult male SD rats were randomly divided into sham operation group (n = 12), modeling group (n = 16), vehicle group (n = 16), SB203580 (a p38 mitogen activated protein kinase [MAPK] inhibitor) treatment group (n = 16) , and MRS1754 (a selective adenosine A2B receptor [A2B AR] antagonist) treatment group (n = 16) according to the random number table. A suture-occluded method was used to induce a middle cerebral artery ischemia-reperfusion model. Vehicle, SB203580, and MRS1754 were injected into the lateral ventricles 30 min before model preparation, the neurological function score was performed after ischemia-reperfusion for 24 h. 2,3,5 triphenyltetrazolium staining was used to detect the infarct volume. The water content of brain tissue was detected by dry-wet weight method. Western blot analysis was used to detect the expression of nSMase 2 and p38 MAPK in ischemic brain tissue. Immunohistochemical staining was used to detect the expression of nSMase 2 in ischemic brain tissue. Results MRS1754 significantly decreased neurobehavioral score (P < 0. 05) and reduced cerebral infarction volume (P < 0. 05) in rats. Both MRS1754 and SB203580 significantly decreased the water content of ischemic brain tissue (all P < 0. 05). In addition, MRS1754 also significantly decrease the phosphorylation of p38 MAPK after ischemia-reperfusion and decreased the expression level of nSMase 2 (P < 0. 01). Conclusion Regulation of A2BAR and p38 MAPK of nSMase upstream may play a neuroprotective role after cerebral ischemia-reperfusion injury.

Effect of adenosine A2A receptor antagonists on motor disorders induced by 6-hydroxydopamine in rat

PURPOSE: To investigate the role of adenosine A2A receptors on 6-OHDA-induced motor disorder in rat. METHODS: In order to induce experimental model of Parkinson's disease, 6-hydoxydopamine (8 μg/rat) was injected unilaterally into the SNc. After three weeks as a recovery period, 6-OHDA-induced bradykinesia and balance disturbances were assessed by using beam traversal test 10, 30 and 60 minutes after intraperitoneal injections of the drugs (caffeine, SCH58261). RESULTS: The results showed that 6-OHDA (8 μg/rat, Intra-SNc) induced motor disorders of Parkinson's disease and increased elapsed time in the beam test (p<0.001). Injection of caffeine (30 mg/kg, i.p.) and SCH58261 (2 mg/kg, i.p.) attenuated elapsed time on beam (p<0.01 and p<0.001). We showed that acute administration of caffeine and SCH 58261 can improve the 6-OHDA-induced bradykinesia and motor disturbance. CONCLUSION: Adenosine A2AR antagonists improve 6-OHDA-motor deficit and this effect seems to be mediated by the inhibition of A2A presynaptic receptors in substantia nigra pars compacta.

Effects of Adenosine and the Selective Adenosine Receptor Agonist on the Expression of Interleukin-18 in Patient with Unstable Angina / 天津医药

Objective To observe the effects of adenosine and adenosine receptor agonist on the expression of inter?leukin-18 (IL-18) in patients with unstable angina (UAP), and the mechanism of adenosine receptor agonists thereof. Meth?ods Fifteen UAP and 15 healthy volunteers were included in this study. The effects of adenosine and the selective adenos?ine receptor agonists on the expression of IL-18 were measured by enzyme-linked immunosorbent assay (ELISA). Results The lower concentration of adenosine (1-100μmol/L) increased the expression of IL-18 in UAP group;whereas the higher concentration of adenosine (1 mmol/L) inhibited the expression of IL-18. The adenosine A1 receptor agonist and A3 receptor agonist increased the expression of IL-18, while the adenosine A2a receptor agonist inhibited the expression of IL-18. There was no significant effect for A2b receptor agonist on the expression of IL-18. Conclusion The lower concentrations of ade?nosine can enhance the expression of IL-18 through adenosine A1 and A3 receptors in UAP. The higher concentrations of ade?nosine can inhibit the expression of IL-18 through adenosine A2a receptors. Adenosine can promote IL-18 expression in lower concentrations.

Adenosine A2A receptor and nervous system diseases / 国际药学研究杂志

Adenosine, which arises from the breakdown of adenosine triphosphate, is extensively distributed in mammalian tissues and cells. Adenosine exerts its regulating effects on cell function via binding to the specific membrane receptors. Adenosine receptors are belong to G-protein-coupled receptors and can be subdivided into Ai, A2A, A2B and A3 receptors in mammals. Among these receptors, A2A has been demonstrated to be related to pathogenesis of many diseases. In the present review, we focus on the recent progress made in investigating the relationship between A2A and nervous system disorders, which may provide new strategies for treatment of these diseases.

Adenosine A2A receptor and nervous system diseases / 国际药学研究杂志

Adenosine, which arises from the breakdown of adenosine triphosphate, is extensively distributed in mammalian tis-sues and cells. Adenosine exerts its regulating effects on cell function via binding to the specific membrane receptors. Adenosine receptors are belong to G-protein-coupled receptors and can be subdivided into A1, A2A, A2B and A3 receptors in mammals. Among these receptors, A2A has been demonstrated to be related to pathogenesis of many diseases. In the present review, we focus on the recent progress made in investigating the relationship between A2A and nervous system disorders, which may provide new strategies for treatment of these diseases.

Effect of adenosine A2A receptor on stress response of pituitary-adrenal axis in acute phase of craniocerebral trauma in mice / 中华创伤杂志

Objective To investigate the effect of adenosine A2A receptor on pituitary-adrenal axis response in acute phase of moderate craniocerebral trauma.Methods Eighteen adenosine A2A receptor knock-out mice in a C57BL/6 background and another eighteen their wild-type littermates were divided into normal control group and craniocerebral trauma for 4 hours group,and craniocerebral trauma for 24 hours group according to random number table,with siμ mice per group.Plasma levels of adrenocorticotropic-hormone (ACTH) and corticosterone at hours 4 and 24 postinjury were determined using ELISA method.Results At 4 and 24 hours,brain water content in wild-type mice [(80.950 ± 0.184) ％,(82.178 ± 0.255)％ respectively] was higher than that in gene knock-out mice [(80.006 ± 0.199)％,(81.091 ± 0.295)％ respectively,P ＜ 0.01].Besides,brain water content in both wild-type and gene knock-out mice increased after injury (P ＜ 0.01).Plasma levels of ACTH and corticosterone were higher in geneknock-out sham mice than in wild-type sham mice [(120.214 ± 2.472) ng/L vs (91.767 ±7.395) ng/L,(27.814 ±0.888) μg/L vs (11.430 ±0.644) μg/L respectively,P ＜0.0l].At 4 and 24 hours,plasma levels of ACTH [(174.776-± 5.040) ng/L,(189.613 ± 4.802) ng/L respectively] in geneknock-out mice showed a higher increase than those in wild-type mice [(119.594 ± 6.945) ng/L,(124.93-± 11.001 7) ng/L respectively,P ＜ 0.05].Moreover,plasma levels of corticosterone [(40.138 ±-0.805) μg/L] at 4 hours and [(37.440-0.485)μg/L] at 24 hours in gene knock-out mice showed a same result as compared with that in wild-type mice [(19.702 ± 0.804) μg/L,(17.602 ± 0.743) μg/L respectively,P ＜ 0.05].Conclusions Knock-out of adenosine A2A receptor increases the release of ACTH and corticosterone in acute stage of moderate craniocerebral trauma and promotes pituitary-adrenal stress response.This may provide a novel explanation for the neuroprotective effect of A2A receptor deficiency.

The effects of adenosine A2A receptor antagonist on the behavior and the striatal A2A receptor,metabotropic glutamate receptors protein expression in rats with levodopa-induced dyskinesia / 中华神经科杂志

Objective To study the behavioural changes and biological effects of selective adenosine A2A receptor antagonist (CSC) in a rat model of levodopa(L-DOPA) -induced dyskinesia (LID).Methods The hemi-parkinsonian rat model was produced by stereotaxically injecting 6-OHDA to the right medial forebrain bundle. Rats were randomly divided into 4 treatment groups with a random number generating program to receive intraperitoneal injections twice daily for 21 days (n = 10): saline, L-DOPA at 25 mg/kg with benserazide at 6. 25 mg/kg, CSC at 2. 5 mg/kg alone and CSC at 2.5 mg/kg with L-DOPA at 25 mg/kg plus benserazide at 6. 25 mg/kg. Forepaw adjusting steps, abnormal involuntary movements (AIM) and rotational response duration were observed on 2, 9, 11,18 and 21 d. After sacrifice, the expression of adenosine A2A R and mGluR5 was observed by Western blot. Results Co-administration of LDOPA with CSC significantly increased the forelimb adjusting steps of parkinsonian rats during 21 days of treatment when compared to L-DOPA alone. CSC treatment alone increased the forelimb adjusting steps significantly. Co-administration of L-DOPA with CSC ( ( 11 ± 5 ) score) significantly decreased the AIM scores of limb and orolingual muscles when compared to L-DOPA alone (( 17 ± 4) score; t = 2. 44, P ＜0. 05). The subchronic L-DOPA treatment upregulated the striatal expression of adenosine A2A R and mGluR5. However, co-administration of L-DOPA with CSC reversed the shortening of the rotational motor response duration induced by L-DOPA administration during the period of the treatment and attenuated the LDOPA-induced upregulation of adenosine A2A R and mGluR5 expressions. Conclusions CSC improves motor function in a hemi-parkinson rat model, potentiates the antiparkinsonian effects with L-DOPA and partly attenuates LID. Co-administration of L-DOPA with CSC reverses the L-DOPA-induced upregulated expression of A2A R and mGluR5, indicating the involvement of both A2A R and mGluR5 in the onset and progression of LID. Adenosine A2AR antagonists may be promising drugs for treatment of LID.

Role of A2B adenosine receptor in 6% HES 130/0.4-induced reduction of pulmonary capillary permeability in a rat model of sepsis / 中华麻醉学杂志

Objective To investigate the role of A2B adenosine receptor(A2BAR)in 6% HES 130/0.4-induced reduction of pulmonary capillary permeability in a rat model of sepsis.Methods Fifty male SD rats weighing 250-300 g were randomly divided into 5 groups(n = 10 each): group Ⅰ sham operation(group S);group Ⅱ sepsis(group CLP);group Ⅲ ,Ⅳ,Ⅴ low,medium,high dose HES(group H1,2,3).The animals were anesthetized with intraperitoneal pentobarbital sodium 50 mg/kg.Left carotid artery and left femoral vein were cannulated for MAP and HR monitoring and fluid and drug administration.Sepsis was induced by cecal ligation and puncture (CLP).6% HES 130/0.4 7.5,15.0 and 30.0 ml/kg were infused iv over 2 h in group H1,2,3 respectively at 4 h after CLP.The animals were sacrificed at 6 h after CLP.The lungs were isolated for determination of pulmonary capillary permeability(by iv Evans blue injection),the expression of A2BAR and the contents of cAMP,protein kinase A(PKA),TNF-α,IL-6 and IL-10 in the lung tissue.Results CLP significantly increased pulmonary capillary permeability,A2BAR expression and cAMP,IL-6 and TNF-α contents in the lung tissue in group Ⅱ as compared with group S.0.6% HES 130/0.4 significantly reduced pulmonary capillary permeability,increased A2BAR expression,cAMP,PKA and IL-10 and decreased IL-6 and TNF-αcontents in the lung tissue in group H1,2,3 as compared with group CLP.6% HES 130/0.4 decreased pulmonary capillary permeability and up-regulated A2BAR expression in a dose-dependent manner.6% HES 130/0.4 15.0 ml/kg was most effective in increasing cAMP and PKA contents in the lung and depressing inflammatory response.Conclusion 6% HES 130/0.4 decreases pulmonary capillary permeability in a rat model of sepsis by up-regulating A2BAR expression in lung tissue.

The dynamic changes of adenosine receptors imbalance during kindling by lithium chloride-pilocarpine / 中华神经科杂志

Objective To investigate the dynamic changes of adenosine receptors, A1 (A1R) and A2a (A2aR) in the brain from the acute to chronic phase after kindling and to explore the correlation between seizure and expression level of A1R and A2aR. MethodsRats were randomly selected into the testing model, reference and normal control groups.Testing rats were kindled by lithium choride-pilocarpine, reference rats were treated with saline, and no treatment was given in normal control group. The dynamic expression of A1R and A2aR were detected by RT-PCR, immunofluorescence staining and Western blot at time-points of 24 hours, 1 month and 6 months post-kindling. Results In the acute phase of 24 hours after kindling, the A1R expression level (mRNA level was (1. 1483 ±0. 1182); Western blot result was ( 0. 7872± 0. 0621 ) ; immunofluorescence staining count was ( 76. 17 ± 4. 62 )/HP) was increased and A2aR (mRNA level was (0. 8338±0. 0572) ; Western blot result was (0. 2098 ±0. 0257) ; immunofluorescence staining count was (43. 83 ± 5. 12 )/HP) was decreased.The results showed statistically difference compared with the reference and normal groups (P< 0. 05 ). In the later chronic phase of 1 month and 6 months after kindling, the expression level of A1R was decreased and A2aR was increased. These data revealed statistically significant difference (P <0. 01 ). Furthermore, the comparison of the results in 1 month and 6 months after kindling found that the expression of AIR was lower in 6 months (mRNA level was (0. 5682 ±0. 0443) ; Western blot result was (0. 7749 ±0. 0262) ; immunofluorescence staining count was (38. 50 ±4. 81 )/HP) than in 1 month and that of A2aR was higher in 6 months (mRNA level was (1. 2169±0. 0332) ; Western blot result was (0. 7080 ±0. 0371 ); immunofluorescence staining count was (114. 50 ± 4. 04)/HP). The differences were statistical significant (t = - 19. 02--13.28, P < 0. 05). ConclusionsThe expressions of A1R and A2AR during and after kindling presents a bidirectional change. In the acute phas the expression of AR is regulated to suppress seizures. While in the chronic phase, the repeated seizures result in the change of A1R and A2aR expression in the opposite direction. This mechanism plays an important role in refractory seizures.

Adenosine A2A receptor antagonist 8-( 3-Chlorostyryl) caffeine improves motor complication in Parkinson's disease / 中华神经科杂志

Objective To investigate cellular and behavioral effects of adenosine A2A receptor antagonist in a rat model of levodopa-induced motor complications.Methods The hemi-parkinsonian rat model was produced by stereotaxically injecting 6-OHDA to right medial forebmin bundle(MFB).Animals were intraperitoneally treated with levodopa 50 mg/kg plus benserazide 12.5 mg/kg twice a day for 22 days levodopa + vehicle.Rotational duration was estimated.After they were sacrificed,the expression of adenosine A2A receptor was observed by immunohistochemistry and Western blot.Results CSC,reversing the shortened rotational duration induced by levodopa,prolonged the rotational duration.This effect was maintained fil the end of the treatment.The chronic levedopa treatment induced an upregulation of adenosine A2A receptor expression in the lesioned striatum (IOD,(11.55±2.75)×104).The subsequent CSC treatment decreased the adenosine A2A receptor expression to the level of control (IOD,(6.02±1.29)× 104) and PD group (IOD,(5.60±1.83)×104>,F=33.31,P<0.05).Conclusion These results suggest that adenosine A2A receptor is probably involved in the development of levodopa induced motor complications and adenosine A2A receptor antagonist could be useful in the treatment of motor comphcations in parkinsonian patients.

Activation of adenosine receptor A2a up-regulates epithelial sodium channel ?-subunit in cultured alveolar epithelium cells / 第三军医大学学报

Objective To investigate the effects of adenosine receptor A2a on the expression of epithelial sodium channel ?-subunit (?-ENaC) in alveolar epithelium A549 cells and the effects of adenosine receptor A2a in acute lung injury/aute respiratory distress syndrome. Methods After alveolar epithelium A549 Cells were incubated with 0,0.1,1,10 and 100 ?mol/L adenosine receptor A2a agonist CGS-21680 for 8 h or with 100 ?mol/L CGS-21680 for 0,1,4,8,24 and 48 h respectively,the mRNA and protein levels of ?-ENaC were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis respectively. Results After A549 cells were incubated with CGS-21680 at different doses for 8 h,the mRNA and protein levels of ?-ENaC were elevated significantly at 0.1 ?mol/L CGS-21680 treatment compared with the control group (P

Effects of selective deletion of adenosine A_(2A) receptors of peripheral leukocytes on cerebral white matter injury induced by chronic hypoperfusion / 解放军医学杂志

Objective To investigate the effects of deletion of adenosine A2A receptors on peripheral leukocytes on cerebral white matter lesions induced by chronic cerebral hypoperfusion.Methods Twenty-four wild type(WT) male mice were given a ? irradiation of 12.5Gy,followed by receiving bone marrow cells tail vein from female A2A receptor knocked out(KO) mice via tail vein,were assigned as KO→WT group,while those received bone marrow cells from WT female mice were assigned as WT→WT group(n=20).The efficiency of reconstitution of bone marrow cells in recipient mice was assessed 7 weeks after transplantation by PCR and immunofluorescent technique.Then,the recipient mice were subjected to bilateral common carotid artery stenosis with internal diameter of 0.18mm by external banding using microcoils at 8 weeks after transplantation.On 7d,14d and 30d after the surgery,corpus callosum,fiber bundles of Caudoputamen and optic tract were harvested from the cerebral white matter,and stained with Kluver-Barrera staining for observing the changes in nerve fibers,and with GFAP and CD11b immunohistochemistry staining for observing the proliferation of microglia and astrocytes.Results At 7 weeks after successful transplantation,the genotype of sex chromosome in peripheral leukocytes of the male recipient mice was changed into female pattern.The expression rate of A2A receptor was 9.73%?2.05% in KO→WT group and 93.82%?11.24% in WT→WT group,with significant difference between the two groups(P