Effects Of Methylprednisolone On INF-Y And AQP4,AQP5 In Tuberculous Pleural And Its Therapeutic Effect

Objective:To construct a rabbit model of tuberculous pleural effusion and observe aquaporin 4(AQP4)and aquaporin 5(AQP5)in the visceral pleura and parietal pleura of tuberculous pleural effusion rabbit model's Dynamic expression on tissue;and detect interferon-y(INF-?)concentration by ELISA method,observe the dynamic changes of INF-y during the onset of tuberculous pleural effusion;methylprednisolone will be injected into the pleural cavity To investigate the effect of it on the concentration of INF-y in tuberculous pleural effusion and the expression of AQP4 and AQP5 on the pleura of rabbits,as well as the therapeutic effect on tuberculous pleural effusion.Methods:Forty-two New Zealand white rabbits were randomly divided into two groups,21 in the control group and 21 in the experimental group(methylprednisolone injection group).Rabbits were injected with Mycobacterium tuberculosis suspension into the pleural cavity to establish the model of TPE.The chest B-ultrasound was performed during the modeling process to observe the pleural effusion and adhesion,and the formation of fiberboard.After the rabbit model was successfully established,the pleural effusion was take once for each of the two groups in the 1st,3rd,5th,7th,10th,15th 20th stages,3 animals were sacrificed and the pleural tissue of rabbit was taken at the same time;meanwhile,the methylprednisolone injection(dose:10mg/kg)was injected into the pleural cavity of the remaining surviving rabbits in the experimental group.ELISA was used to detect the concentration of INF-y in each pleural effusion and observe the dynamic changes of the pleural effusion;The expression of AQP4 and AQP5 in the removed visceral and parietal pleura tissue was detected by Western blot;the expression of AQP4 and AQP5 in pleural tissue was detected by immunohistochemical staining;the mRNA expression of AQP4 Gene and AQP5 Gene in rabbit pleural tissue were detected by RT-PCR.Results:1.Changes in pleural effusion content:The pleural effusion content of the rabbit model in the control group can be seen to increase significantly with the increment of time,reaching the highest point when it reaches 15th days,and it display the downtrend from 15 to 20 days.experimental group,the rabbits after 10 days the volume of the pleural effusion in the model decreased and returned to a similar level at the 20th day as at the 5th day.2.The changes of INF-y concentration in pleural effusion:detected by ELISA method,in the control group:from 1 to 20 day,with the increment of time,the concentration of INF-yin pleural effusion showed a significant increasing trend;On the 20 th day,the INF-y concentration in the pleural effusion of the rabbit model reached its peak.;however,the INF-y concentration in the pleural fluid showed a significant decrease from 7 to 20 days,and the The downward trend was more obvious from 7 to 10 days.During the first 7 days,the inf-? concentration in the pleural effusion of the two groups showed a significant increasing trend with the increment of time,but each time The detection of the experimental group was lower than the INF-y concentration in the pleural fluid of the control group by comparing the experimental group with the control group.On the 10th?15th and20th day,the control group was compared with experimental group.The difference of INF-y concentration in the pleural effusion of the rabbit animal model was statistically significant(p<0.05).3.AQP4 and AQP5 expression on pleural tissue:Western blot and immunohistochemical staining revealed that the expression of AQP4 and AQP5 in the cytoplasm and nucleus of visceral pleural cells was higher than that in the parietal pleural.After comparing the experimental group with the control group,methylprednisolone has a certain inhibitory effect on the expression of AQP4 and AQP5 in pleural tissue.4.AQP4mRNA and AQP5mRNA expression on pleural tissue:RT-PCR experiments showed that the expression of AQP4mRNA and AQP5mRNA on pleural tissue in the experimental group was higher than that in control group.AQP5 mRNA expression is low.Conclusions:1.Injecting Mycobacterium tuberculosis suspension into the pleural cavity of rabbits can successfully establish a model of tuberculous pleural effusion in rabbits.2.It shows methylprednisolone has a certain inhibiting effect on tuberculous pleural effusion.3.Methylprednisolone can inhibit the expression of AQP4 and AQP5 on parietal and visceral pleura.4.The expression of AQP4 and AQP5 on visceral pleura is more abundant than that in parietal pleura5.Methylprednisolone can reduce the concentration of INF-? in pleural effusion,and the effect is more obvious after the 7th day of administration.