| Object iveBy copying the inflammatory pleural effusion in a rat model, selected the passage of water and ion channels as targets, then observed their effection in the forming process of Xuanyin(pleural effusion), To elucidate the pathological mechanism,of pleural fluid formation, At the same time observe the therapeutic effect of ten jujubes Decoction on a rat model of Xuanyin. Through the comparison of ten jujube soup before and after treatment, dynamic observation of changes of the indexes, From the water and ion channel approach to discuss pleural effusion formation mechanisms in the pathophysiology and ten jujubes decoction treatment mechanism.Methods1. Experimental animal and grouping:Seventy-two male healthy Wistar rats were randomly divided into normal group(8), model group (32)and treatment group(32). Model group and treated group according to the injection of carrageenan after execute on time was divided into4pleuritis groups (12,24,36, and48h groups respectively)2.The method of making model:Replication the rat model of inflammatory pleural effusion as Xuanyin model, model making reference to Cuzzocrea method. In10%chloral hydrate abdominal anesthetized rats,75%alcohol disinfection, on the right chest wall midclavicular line corresponds to the breast at the thoracic puncture, model group, treatment group of intrapleural injection of1%carrageenan solution0.2mL, control group was injected with an equal volume of saline.3.Treatment:Molding is completed, the treatment group were given ten jujube Decoction according to10mL/kg/time in the model after6hours and10hours, model group at the same time gavage of equal volumes of saline.4. Materials and making samples:Rats were executed respectively by abdominal aorta bloodletting after12h24h36h and48h injected in harpoons food glue. The control group were executed after24hours injected in physiological saline. Observe the distribution of pleural effusion and pleural adhesions degree after the exposure of bilateral pleural by the way of cutting off the skin and the sternum from the xiphoid. The bilateral pleural exudate were drained which were measured accurately by lmL syringe and in which white blood were counted and classified by the conventional approach. The wall layer and visceral pleura specimens were taken and then part of the organization was fixed to the10%neutral Formalin liquid and the rest used for the extraction of total RNA and protein were conserved in-80℃fridge.5. Biopsy:embedded in paraffin, sliced, slice thickness3um, HE staining, To observe the degree of pleural inflammation using the microscope.6. Determined the AQP1, ENaC, CFTR protein expression levels in Pleural tissue by immunohistochemical method:Operated in accordance with the general steps7. Determined the AQP1, ENaC, CFTR gene expression levels in Pleural tissue by RT-PCR:Operated in accordance with the general steps8. Statistical analysis:SPSS13.0statistical package was used to analyze the data, P<0.05was considered statistically different.Results1. Animals generallyNormal rats:rats were lively, responsive, with shiny hair, appetite, urined and defecate was normal, no abnormal signs such as cough, shortness of breath.Model rats:the spirit of rats was sluggish, unresponsive, hairwas withered yellow, poor appetite, and the prevalence of shortness of breath, and occasionally you could hear the coughTreated rats:the spirit of rats somewhat less, reflection was relat ively slow, loss of appetite compared with model group; loss of appetite compared with the model group;stool was slightly runny, urine volume in creased, a slight shortness of breath.2. Analysis of the pleural effusion:carrageenan injection group coul d be seen the pale yellow pleural effusion, and occasional pleural adhesi ons. The pleural cavity of the model rats injected carrageenan exudate, t he volume of pleural effusion reached the peak in24hour, began to declin e after36hours;until48hours only a small amount of liquid exudation was left; treatment group also had a similar law, but at the same timepoi nt the volume of pleural effusion had a significant decrease compared to the model group; the control group had no pleural effusion. Pleural fluid collected from the chest cavity took to do a routine inspection, The re sults showed that the WBC count of pleural effusion was elevated and main ly multinucleated cells, The treatment group decreased compared to the mo del group, but. no statistically significant difference (P>0.05)3. Pathologic:Observed under light microscope, HE staining showed th at the control group, pleural skin integrity, was flat, single-layer arra ngement, no interstitial edema and inflammatory cell infiltration. Pleura1effusion model group could be seen that the pleural interstitium was si gnificantly thicker and significant edema, diffuse inflammatory cell infi ltration, showing small abscess formation, some mesothelial cells shed in the region. The treatment group also could be seen pleural interstitial edema, inflammatory cells scattered infiltration, but lighter than the mo del group.4. The expression of AQP1, ENaC and CFTR protein in Rat pleural tis sue test results:Immunohistochemistry showed that the control group and p leurisy rats are dirty, parietal pleural mesothelial cells and capillary endothelial cells showed a brown-yellow staining, indicating the presence of AQP1in, of ENaC and CFTR protein expression. Visceral pleura to expr ess strong expression in the parietal pleura, the model group and treatme nt group than the control group strengthened;24h group the highest expre ssion of the model group, the staining signal was coarse granular, browni sh black. Model group at the same time point the expression intensity tha n the treatment group at the same time point; Comparied the number of pos itive cells of model group and treatment group, the differences were stati stically significant (P<0.055. The expression of AQP1, ENaC and CFTR gene in Rat pleural tissue test results:In pleural tissue of rats of the model groups and treatmen t groups, The amount of AQP1gene expression were increased by a statisti cally significant difference.compared with the normal group, Model grou p increased at12h,24h set of gene expression is highest. And then gradu ally decline. The treatment group also has the same law. Model and treate d groups at the same time pointcompared, gene expression in the model gro up more than in the treatment group, and the difference was statistically;The ENaC and CFTR gene had a similar law.ConclusionTen Jujubes Decorction could facilitate the absorption of pleural effusion in rat model of Xuanyin, and could educe the infiltration of inflammatory cells. the protein expression and gene expression levels of AQP1ENaC and CFTR in the model group was raised compared with treatment groups and norm al group, suggesting that water channels and ion channels participated in th e inflammatory pleural effusion transfer process; At the same time point the treatment groups indicators had a lower expression level than model g roup, suggesting that the Ten Jujubes Decorction could regulate the expression level of the water channels and ion channels, to promote the trans-shipmen t of pleural effusion, and the treatment of Xuanyin(pleural effusion)... |
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