BACKGROUND:It has been confirmed that c-fos expression increased markedly in hypothalamic paraventricular nucleus(PVN)during asthmatic attack in rats,and PVN has extensive physiological functions,involving in the regu...BACKGROUND:It has been confirmed that c-fos expression increased markedly in hypothalamic paraventricular nucleus(PVN)during asthmatic attack in rats,and PVN has extensive physiological functions,involving in the regulation of respiratory system,etc.OBJECTIVE:To observe the alteration of electroencephalogram(EEG)and power spectra in PVN during the asthmatic attack,and the alteration of lung function and diaphragmatic muscle discharge after bilateral PVN lesion in asthmatic rats.DESIGN:A randomized control study.SETTING:Laboratory of Physiology and Pharmacology,School of Basic Medical Sciences,Southeast University.MATERIALS:Forty-eight male adult SD rats of 260-300 g were used.The rats were randomly divided into 6 groups(n=8):control group,asthma group,electrolytic lesion of PVN group,KA-induced lesion of PVN group,sham electrolytic lesion of PVN group and sham kainic acid(KA)-induced lesion of PVN group.KA,chicken ovalbumin and aluminum hydroxide were purchased from American Sigma Company.Bordetella pertussis vaccine(Institute of Biological Products of Shanghai);stereotaxic apparatus(JiangwanⅡ,China);lesion-producing device(YC-2 programmable stimulato,Chengdu Instrument Company);MD2000 signal processing system(Nanjing Medical School);data acquisition system(RM6240B,Chengdu Instrument Company).METHODs:The experiments were carried out in the Laboratory of Physiology and Pharmacology,School of Basic Medical Sciences,Southeast University from January to August in 2006.①Rats except for control group were sensitized with an intraperitoneal injection of 100 mg chicken ovalbumin and 100 mg aluminum hydroxide and Bordetella pertussis vaccine containing 5×10^(9) heat-killed in 1 mL of sterile saline.From the fifteenth to seventeenth days rats received three times aerosolized ovalbumin challenge.In rats of the control group and asthma group three steel electrodes were placed into the left PVN(AP-1.8 mm,LR 0.4 mm,OH-7.9 mm),parietal cortex and subcutaneous tissue in lower limb.Lung function tests were carried out simult展开更多
Objective To compare the quality and quantity of total RNA from different source-original neurons applied in LMPC technique. Methods （ 1 ） Aglient 2100 bioanalyzer and RT-PCR were used to check the concentration and...Objective To compare the quality and quantity of total RNA from different source-original neurons applied in LMPC technique. Methods （ 1 ） Aglient 2100 bioanalyzer and RT-PCR were used to check the concentration and fragmentation of total RNA from unfixed, temporal fixed and fixed 12 h hypothalamus sections; （2）Different neurons of PVN and SON were collected by LMPC, CRH, TRH, AVP, OT mRNA level were measured by RT-PCR; （3）Labeled neurons by injecting CTB into stomach and non-labeled neurons in DMV collected by LMPC were checked for house keeping genes by RT-PCR. Results （ 1 ） Unfixed section had higher concentration and better quality of total RNA compared with fixed sections applied in LMPC ; relative short amplicons such as GAPDH, NSE, MCH and MCAR were successfully obtained from fixed and unfixed and long amplicon of GR can only be obtained from unfixed material; （2） In magnocellular PVN and SON the expressions of AVP and OT were more special than those in the parvocellular PVN. Oppositely, the expressions of CRH, TRH in the parvocellular were more special than the other two ; （3） The expressions of house keeping genes had no significant difference between labeled and non-labeled DMV neurons. Conclusion The quality and quantity of total RNA from unfixed brain tissues were better than fixed tissues applied in LMPC and the CTB tracer which may differentiate neurons had no significant effect on physiology of the neurons applied in LMPC. The results showed that the LMPC technique is suitable for the qualitative and quantitative study on individual neurons at mRNA level.展开更多
The present study was designed to investigate the mechanisms by which P2X7 receptors(P2X7Rs)mediate the activation of vasopressinergic neurons thereby increasing sympathetic hyperactivity in the paraventricular nucleu...The present study was designed to investigate the mechanisms by which P2X7 receptors(P2X7Rs)mediate the activation of vasopressinergic neurons thereby increasing sympathetic hyperactivity in the paraventricular nucleus(PVN) of the hypothalamus of rats with acute myocardial ischemia(AMI). The left anterior descending branch of the coronary artery was ligated to induce AMI in rats. The rats were pretreated with BBG(brilliant blue G, a P2X7R antagonist), nelivaptan(a vasopressin V1b receptor antagonist), or diphenyleneiodonium(DPI) [an nicotinamide adenine dinucleotide phosphate(NADPH)oxidase inhibitor]. Hemodynamic parameters of the heart were monitored. Myocardial injury and cardiomyocyte apoptosis were assessed. In the PVN of AMI rats, P2X7R mediated microglial activation, while reactive oxygen species(ROS) and NADPH oxidase 2(NOX2) were higher than in the sham group. Intraperitoneal injection of BBG effectively reduced ROS production and vasopressin expression in the PVN of AMI rats. Moreover, both BBG and DPI pretreatment effectively reduced sympathetic hyperactivity and ameliorated AMI injury, as represented by reduced inflammation and apoptosis of cardiomyocytes.Furthermore, microinjection of nelivaptan into the PVN improved cardiac function and reduced the norepinephrine(AE) levels in AMI rats. Collectively, the results suggest that, within the PVN of AMI rats, P2X7R upregulation mediates microglial activation and the overproduction of ROS, which in turn activates vasopressinergic neuron V1b receptors and sympathetic hyperactivity, hence aggravating myocardial injury in the AMI setting.展开更多
Objective: To observe the influence of electrolytic destruction of nucleus soli tary tract (NTS) and hypothalamic paraventricular nucleus (PVN) on the effect of electroacupuncture (EA) in improving ischemic myocardia ...Objective: To observe the influence of electrolytic destruction of nucleus soli tary tract (NTS) and hypothalamic paraventricular nucleus (PVN) on the effect of electroacupuncture (EA) in improving ischemic myocardia cellular transmembrane action potential (TMAP). Methods: 38 Japanese breed big-ear wh ite rabbits (ane sthetized with 20% Urethane, 4 mL/kg) were randomly divided into acute myocardia l ischemia (AMI) group (n=8), PVN destruction group ( n=12) and PVN+NTS destructio n group (n=18). AMI model was established by occlusion of the descending anterio r branch (DAB) of the coronary artery. TMAP of myocytes was recorded by using a glass microelectrode which was fixed to a suspending spring silver wire. Bilater al "Neiguan"(PC 6) in all the 3 groups were punctured and stimulated electri call y by using parameters of continuous waves, frequency of 7 Hz, intensity of 6 mA a nd duration of 30 minutes. Results: After AMI, ECG-ST elevated significantly whil e APA lowered, APD50 and APD90 shortened clearly in comparison with those of pre -AMI in the 3 groups. Compared with AMI group, ECG-ST values of PVN destructi on group and PVN+NTS destruction group were significantly higher (P <0.05～0.01), whi le APA, APD50 and APD90 all significantly lower in all the recording time course s(P<0.05). The facts displayed that electrolytic destru ction of PVN and PVN+NT S could produce ischemic myocardial injury and reduce the protective effect of E A on ischemic myocardial cells. Comparison between PVN destruction and PVN+NTS g roups showed that all the 4 indexes of the later group were evidently worse than those of the former group (P<0.05), suggesting after des truction of these two n uclei, the effect of EA was worsened further. Conclusion: Electrolytic destru ction of PVN and NTS weakens the protective effect of EA on ischemic myocardial cells, both NTS and PVN take part in the effect of EA of "Neiguan"(PC 6) Point i n improving ischemic myocardium.展开更多
基金the Scientific Foundation of the Ministry of Railway,No.6747600045
文摘BACKGROUND:It has been confirmed that c-fos expression increased markedly in hypothalamic paraventricular nucleus(PVN)during asthmatic attack in rats,and PVN has extensive physiological functions,involving in the regulation of respiratory system,etc.OBJECTIVE:To observe the alteration of electroencephalogram(EEG)and power spectra in PVN during the asthmatic attack,and the alteration of lung function and diaphragmatic muscle discharge after bilateral PVN lesion in asthmatic rats.DESIGN:A randomized control study.SETTING:Laboratory of Physiology and Pharmacology,School of Basic Medical Sciences,Southeast University.MATERIALS:Forty-eight male adult SD rats of 260-300 g were used.The rats were randomly divided into 6 groups(n=8):control group,asthma group,electrolytic lesion of PVN group,KA-induced lesion of PVN group,sham electrolytic lesion of PVN group and sham kainic acid(KA)-induced lesion of PVN group.KA,chicken ovalbumin and aluminum hydroxide were purchased from American Sigma Company.Bordetella pertussis vaccine(Institute of Biological Products of Shanghai);stereotaxic apparatus(JiangwanⅡ,China);lesion-producing device(YC-2 programmable stimulato,Chengdu Instrument Company);MD2000 signal processing system(Nanjing Medical School);data acquisition system(RM6240B,Chengdu Instrument Company).METHODs:The experiments were carried out in the Laboratory of Physiology and Pharmacology,School of Basic Medical Sciences,Southeast University from January to August in 2006.①Rats except for control group were sensitized with an intraperitoneal injection of 100 mg chicken ovalbumin and 100 mg aluminum hydroxide and Bordetella pertussis vaccine containing 5×10^(9) heat-killed in 1 mL of sterile saline.From the fifteenth to seventeenth days rats received three times aerosolized ovalbumin challenge.In rats of the control group and asthma group three steel electrodes were placed into the left PVN(AP-1.8 mm,LR 0.4 mm,OH-7.9 mm),parietal cortex and subcutaneous tissue in lower limb.Lung function tests were carried out simult
文摘Objective To compare the quality and quantity of total RNA from different source-original neurons applied in LMPC technique. Methods （ 1 ） Aglient 2100 bioanalyzer and RT-PCR were used to check the concentration and fragmentation of total RNA from unfixed, temporal fixed and fixed 12 h hypothalamus sections; （2）Different neurons of PVN and SON were collected by LMPC, CRH, TRH, AVP, OT mRNA level were measured by RT-PCR; （3）Labeled neurons by injecting CTB into stomach and non-labeled neurons in DMV collected by LMPC were checked for house keeping genes by RT-PCR. Results （ 1 ） Unfixed section had higher concentration and better quality of total RNA compared with fixed sections applied in LMPC ; relative short amplicons such as GAPDH, NSE, MCH and MCAR were successfully obtained from fixed and unfixed and long amplicon of GR can only be obtained from unfixed material; （2） In magnocellular PVN and SON the expressions of AVP and OT were more special than those in the parvocellular PVN. Oppositely, the expressions of CRH, TRH in the parvocellular were more special than the other two ; （3） The expressions of house keeping genes had no significant difference between labeled and non-labeled DMV neurons. Conclusion The quality and quantity of total RNA from unfixed brain tissues were better than fixed tissues applied in LMPC and the CTB tracer which may differentiate neurons had no significant effect on physiology of the neurons applied in LMPC. The results showed that the LMPC technique is suitable for the qualitative and quantitative study on individual neurons at mRNA level.
基金supported by grants from the National Natural Science Foundation of China (31271215, 81770423, 81973945, and 81673766)the Health Vocational and Technical Education Research Program of Jiangsu Province, China (J201506)。
文摘The present study was designed to investigate the mechanisms by which P2X7 receptors(P2X7Rs)mediate the activation of vasopressinergic neurons thereby increasing sympathetic hyperactivity in the paraventricular nucleus(PVN) of the hypothalamus of rats with acute myocardial ischemia(AMI). The left anterior descending branch of the coronary artery was ligated to induce AMI in rats. The rats were pretreated with BBG(brilliant blue G, a P2X7R antagonist), nelivaptan(a vasopressin V1b receptor antagonist), or diphenyleneiodonium(DPI) [an nicotinamide adenine dinucleotide phosphate(NADPH)oxidase inhibitor]. Hemodynamic parameters of the heart were monitored. Myocardial injury and cardiomyocyte apoptosis were assessed. In the PVN of AMI rats, P2X7R mediated microglial activation, while reactive oxygen species(ROS) and NADPH oxidase 2(NOX2) were higher than in the sham group. Intraperitoneal injection of BBG effectively reduced ROS production and vasopressin expression in the PVN of AMI rats. Moreover, both BBG and DPI pretreatment effectively reduced sympathetic hyperactivity and ameliorated AMI injury, as represented by reduced inflammation and apoptosis of cardiomyocytes.Furthermore, microinjection of nelivaptan into the PVN improved cardiac function and reduced the norepinephrine(AE) levels in AMI rats. Collectively, the results suggest that, within the PVN of AMI rats, P2X7R upregulation mediates microglial activation and the overproduction of ROS, which in turn activates vasopressinergic neuron V1b receptors and sympathetic hyperactivity, hence aggravating myocardial injury in the AMI setting.
基金This studyis subsidized by National Science Foundation of China (No .30171179) .
文摘Objective: To observe the influence of electrolytic destruction of nucleus soli tary tract (NTS) and hypothalamic paraventricular nucleus (PVN) on the effect of electroacupuncture (EA) in improving ischemic myocardia cellular transmembrane action potential (TMAP). Methods: 38 Japanese breed big-ear wh ite rabbits (ane sthetized with 20% Urethane, 4 mL/kg) were randomly divided into acute myocardia l ischemia (AMI) group (n=8), PVN destruction group ( n=12) and PVN+NTS destructio n group (n=18). AMI model was established by occlusion of the descending anterio r branch (DAB) of the coronary artery. TMAP of myocytes was recorded by using a glass microelectrode which was fixed to a suspending spring silver wire. Bilater al "Neiguan"(PC 6) in all the 3 groups were punctured and stimulated electri call y by using parameters of continuous waves, frequency of 7 Hz, intensity of 6 mA a nd duration of 30 minutes. Results: After AMI, ECG-ST elevated significantly whil e APA lowered, APD50 and APD90 shortened clearly in comparison with those of pre -AMI in the 3 groups. Compared with AMI group, ECG-ST values of PVN destructi on group and PVN+NTS destruction group were significantly higher (P <0.05～0.01), whi le APA, APD50 and APD90 all significantly lower in all the recording time course s(P<0.05). The facts displayed that electrolytic destru ction of PVN and PVN+NT S could produce ischemic myocardial injury and reduce the protective effect of E A on ischemic myocardial cells. Comparison between PVN destruction and PVN+NTS g roups showed that all the 4 indexes of the later group were evidently worse than those of the former group (P<0.05), suggesting after des truction of these two n uclei, the effect of EA was worsened further. Conclusion: Electrolytic destru ction of PVN and NTS weakens the protective effect of EA on ischemic myocardial cells, both NTS and PVN take part in the effect of EA of "Neiguan"(PC 6) Point i n improving ischemic myocardium.
