Effect And Mechanism Research Of Paeonal On Bronchialasthma Rats

Objective: To explore the mechanism of Paeonal on bronchial asthma based on PI3K signaling pathway,to provide experimental basis for its clinical application,by taking bronchial asthma model rats as the research objects.Methods:60 SD male rats were fed adaptation for a week,10 randomly selected for normal control group,asthma model group,aminophylline(27mg/kg)positive control group,high(300mg/kg)Paeonal dose group,medial(150mg/kg)dose group,low(75mg/kg)dose group.In addition to the normal group,Rats were sensitized ten parts with 0.2% of ovalbumin(OVA),1m L/mouse.Including the neck 1,the back of the 3,the two digits of front and behind 4,the groin of both sides 2,every point was subcutaneous injected with 0.2% of OVA,and i.p the last 0.5m L,setting up the model rat of bronchial asthma.Starting from the 15 th day,giving appropriate drugs(10ml/kg)every day(drugs were configured with 0.5% CMC-Na),and normal group and model group were given equivalent 0.5% CMC-Na.Meanwhile,after 0.5h,apart from normal group,each group was attacked 30 minutes by 1% OVA for 4 weeks.Observing the common conditions of rats and weighing every week.After the last attack for 24 h,we took the rat lung,4%neutral poly formaldehyde solution fixed,embedded in paraffin wax.Sections were stained with HE method to observa rats' lung structure,After Ach was injected,detecting and recording the change of airway resistance(AR)and lung compliance(CL)within 5minutes by respiratory function instrument.To calculate the amounts of white blood cells in BALF and its classification with Wright-Giemsa stain method.We took the lungs of rats,weighted and calculated lung index.Last after administration,rats were given for abdominal aortic blood to measure the level of Ig E?IL-4?IL-5?IL-13 and IFN-? with ELISA.And we extraced total protein to detect PI3K protein expression with Western blot.Results:1.Effect of Paeonal on the general signs of bronchial asthma rats.After sensitized with OVA and challenged by nebulized antigen,the rats of model group appearanced the phenomenon of hair sere lodging,restlessness,shortness of breath,abdominal flapping,sneezing,nodding heads,scratching ears,bitting each other,eating less,urinary and fecal incontinence,oral cyanosis,dark purple tongue etc.Compared with model group,the general state of normal control group were normal,the treatment groups were improved,and the body weight of rats in treatment groups had no difference.2.Effect of Paeonal on lung function and airway hyperresponsiveness of asthma rats.Compared with the normal control group,model group rats of airway resistance(AR)were significantly increased and lung compliance(CL)were significantly decreased(P<0.01),besides,with the Ach stimulus concentration gradually increased,lung resistance rising and lung compliance gradually declined.The high,medial and low dose group were all could reduce airway resistance and different level improve lung compliance,including high,middle dose group of the most significant effect.Compared with asthma model group,the treatment groups had a significantly lower asthma rat lung resistance,pulmonary compliance increased significantly,with significant difference(P < 0.05),of which the most significant effect on inhibiting lung resistance increase,and the concentration of acetylcholine stimulation and inhibition of airway hyperresponsiveness were positively correlated.3.Effect of Paeonal on the number of white blood cells and its classification in BALF of asthma rats.Compared with the normal group,model group rats alveolar lavage in total number of white blood cells(5.71�0.70)were significantly increased(P < 0.01),after treatment,total number of white blood cells were significantly lower;Compared with model group,the total number of white blood cells in rat alveolar lavage treatment group were significantly lower,with significant difference(P<0.05,P<0.01).Compared with the normal control group,the count of EOS(9.4�0.30),neutrophil(24.2�0.40)and lymphocyte(55.77�0.35)in BALF asthma model group were significantly higher(P < 0.01),showing that asthma model was successful.Compared with model group,the number of EOS in Pae medial(5.5�0.40),high(6.33�0.45)dose group and aminophylline group(8.0�0.46)were decreased significantly(P < 0.01),suggesting the Pae medial,high dose group could inhibit asthma EOS infiltration of inflammation obviously.4.Effect of Paeonal on the lung weight and lung index of bronchial asthma rats.Compared with normal control group,Asthma model group rats lung index increased significantly,with statistical significance(P < 0.01);But after treatment,lung index were significantly decreased,Pae high and middle dose group compared with model group with statistical difference(P < 0.05).Compared with model group,the lung index of Pae low dose group also reduced,but no statistical difference(P >0.05).Suggesting that Pae can significantly reduced the lung swelling and in high doses(0.53%�0.075%)is preferred.5.Effect of Paeonal on lung tissue morphology in asthma rats.The kind of pathological staining with HE showed that normal tracheal mucosa epithelial integrity,pulmonary alveoli structural normal,bronchial wall thickening was not obvious,and no inflammatory cell infiltration.Model group submucosal bronchial wall and around the wall had a lot of inflammatory cell infiltration,bronchial lumen narrowing,and bronchial wall thickening.Compared with the normal control group,bronchial wall thickening obviously and airway smooth muscle hyperplasia.Compared with the model group,the high Paeonal dose group,medial dose group and low dose group inflammatory cell infiltration were significantly decreased,bronchial wall and pipe wall smooth muscle became thin.6.Effect of Paeonal on the serum Ig E,IL-4,IL-5,IL-13,IFN-? of treating bronchial asthma rats.The levels of serum Ig E(22.50�0.96),IL-4(116.12�15.01),IL-5(85.52�6.85)and IL-13(45.03�2.77)in asthma model group rats were significantly increased compared with normal control group(P < 0.01),while IFN-?(89.92�6.16)were significantly reduced(P < 0.01);rats in the treatment groups compared with model group,the level of serum Ig E,IL-4,IL-5 and IL-13 were decreased significantly(P<0.05,P<0.01),and IFN-? were significantly increased(P<0.05,P<0.01).7.Effect of Paeonal on expression of PI3K protein in treating bronchial asthma rats.Compared with the normal group,the level of PI3K protein expression in model group rats(2.20�0.05)were significantly increased(P < 0.01),compared with the model group,Pae high,medium and low dose group and positive control group,PI3K protein expression appeared different degree of decline(P < 0.01).Conclusion: Paeonal has significant effect of bronchial asthma.It can significantly improve the index of lung function in asthma rats,delay the pathological lesions in the lung tissue,reduce the infiltration of inflammation cells,gas pipe wall thickening and airway smooth muscle hyperplasia,the prevention and control of bronchial asthma have a certain effect,and its mechanism may be related to affecting the PI3K signaling pathway and the expression of PI3K protein.