Sd Rats Were The Package Qinge Complex Microinjection Of Asarum Including The Respiratory Effect Of Serum

ObjectiveTo observe effects of microinjection of serum containing Asarum into pre-Botzinger Complex on respiration in adult rats.Methods1.Make the blood serum containing AsarumFirst prepares certain Asarum root powder of density give SD rat to irritated stomach,twice a day,for eight days.The dosage of Asarum were according to The Pharmacopeia of the People's republic of China(lst part,public in 2005).2.Effects of microinjection of serum containing Asarum into pre-Botzinger Complex on respiration in adult ratsSelects certain healthy grown-up SD rats,with the body weight of 250- 350g(no concern of female or male).Does the intraperitoneal injection of anesthesia with Ukrainian Ruttan 20%by 5ml/kg.Does the trachea cannula,separates the two-sided neck fan to be distracted passes through.Opens the abdominal cavity,separates the diaphragm,records the diaphragm electric discharge,and then the diaphragm electric discharge tests the system surveillance or the record by the BL-420 biology function.Through two cock the ears the pole and the tooth hitch fixes the SD rat forehead on the brain three-dimensional position finder, center big mouse cranial vault line arrowy margin,long approximately 2cm,separation Subcutaneous tissue,exposes the before and after fully xin of the skull,adjusts the brain three-dimensional position finder, makes before and after xin in the identical horizontal plane.By Paxinos and Watson big mouse brain three-dimensional localization atlas localization pre-Botzinger Complex(PBC),the locating point is established:12.3mm after the coronal suture,2.2mm away from the median line,10.3mm under the plane of the skull.Drills an eyelet for the center at the intersection point of right flank side 2.2mm and the coronal suture after the skull median line left side of the 12.3mm,fills the 0.2mm microinjector with 0.10μl solution and fixes the syringe outer diameter on the brain three-dimensional position finder pushing ram, according to Paxinos and the Watson big mouse brain three-dimensional localization atlas zero reference point and the PBC locating point,puts in the micro injector the injection spot place above.Then injection is ready and starts the record from then on.The recorded time is 3min;Afterward shuts off the neck two-sided fan to be distracted passes through,continues to record 5min;Finally inserts slowly the micro injector the PBC locating point,after the arriving,impels the injector to pour into slowly 0.10μl the solution,pushes pours with the uniform speed in 60s,keeps the needle for 5min to guarante that the medicine is absorbed completely,latter takes off the needle slowly,records the entire operating process,and records for 60min continually after the entire operating process.The experiment is divided into the blank blood serum control group (n=10),asarum including the medicine blood serum group(n=10),γ-aminobutyric acid(GABA)the group(n=10),D(-)-2-Amino-5-phosphonopentanoic acid(Ap-5)group(n=10),GABA to add the asarum including medicine blood serum group(n=10),Ap-5 to add the asarum to contain the medicine blood serum group(n=10).It injects separately 0.10μl includes 2%pontamine sky-blue solution asarum including the medicine blood serum,the blank blood serum,0.5mol/L GABA, 0.05mol/L Ap-5,0.5mol/L GABA and the asarum and so on compares the configuration including the medicine blood serum compared to the configuration solution and 0.05mol/L AP-5 and the asarum including the medicine blood serum the solution.After each experiment,takes out the entire brain organization,fixed the specimen 10%formalin solution for 48h,soaks it in 4℃20%sucrose PBS fluid until it goes to the bottom,intercepters the brain stem part, freezes the coronal serial section,40μm of thickness,collects the spot slices nearby the injection place,and carries on the neutral red to dye, the movie-making,the optical microscope observation,according to the mark blue spot position,compares Paxious and Watson atlas,the dyeing spot and Paxinos and Watson atlas PBC localization coordinate tallying includes statistics processing.Finally there are 7 groups of blank blood serum which meet standards,8 of the asarum,7 of the GABA including the medicine blood serum group,6 of the Ap-5 groups,7 of GABA to add the asarum,8 of Ap-5 including the medicine blood serum group with asarum including.Analysis targets include:the oscillation amplitude discharged by the intervals diaphragm at various moments,inspiration time interval(TI), expiration time interval(TE)as well as respiratory rate(RF).Calculats the rate of the change of these targets[observing data after th injection comparing data before the injection)/comparing data before the injection×100%],expresses the obtained data with((?)±s)%,uses the SPSS10.0 statistics software to carry on the data processing,carries on own comparison or two sample mean values the T-test,taking P＜0.05 as the significant limit.Conclusion1.The influence of asarum to respiratory center has close relation with the PBC nuclei;2.Asarum on respiratory generated by the inhibitory effect of the mechanism may mainly produce a role through the PBC in the nuclei of GABA receptors;3.The bossibility that asarums influence the respiratory center through other nuclear groups cannot be removed.