| Objective: Establishing a cold astham rat model by ovalbumin(OVA)to explore the intervention of relieving inflammation of bronchial asthma by Pingchuanning.Based on PI3 K regulation of ER-related ROS and the expression of NF-κB signaling-related proteins NOX4,RIG-Ⅰ and Col6 which activated by RIDD-RIG-Ⅰ axis to alleviate asthma airway inflammation,and then to study its mechanism between Pingchuanning and asthma airway inflammation.Method: 105 SPF class healthy male SD rats,aged 6-8 weeks,were randomly divided into 7 groups with 15 rats in each group.The rats were purchased from Jinan Pengyue Experimental Animal Breeding Limited Company.Feeding all rats for a week in order to make them adapted to the environment.In order to sensitize the rats,on day 1 and 8after adaptation,rats in the model group,Pingchuanning high,medium,low dose groups,Guilongkechuanning group,and dexamethasone group were injected with 10% OVA1 m L,the normal group was given equal amounts of saline instead.The rats were sensitized by atomization from day 15 to establish a cold astham model,put rats in the same cold environment,normal group were atomized with normal saline,others were atomized with 1% OVA suspension,30 minutes per day for 7 days.From the 22 nd day,rats in the treatment group were treated with corresponding drugs once a day for 21 consecutive days according to adult relative dose conversion,the normal,model groups were uesd the same amount of normal saline.After the last administration,rats fast for24 hours,then stimulated with 10% OVA,and dissected within 2 hours.The general behavior of rats was observed throughout the experiment.HE staining and PAS staining were used to observe the inflammatory cells and morphological changes in rats lung tissue;ELISA was used to detect IL-13 and IL-25 levels in BALF;RT-q PCR was used to detect the mRNA expression level of NOX4,RIG-Ⅰ and Col6;Western Blot detection of NOX4,RIG-Ⅰ,Col6 protein expression level.Results: 1.General behavior of rats: rats in the normal group developed well during the whole experiment,with moist fur,sensitive response,normal diet,and uniform respiratory rhythm;the rats in the model group were thin,withered,unresponsive,clustered,reduced diet,loose stools,shortness of breath,sneezing,and audible wheezing;all aspects were improved after treatment in each medication group.2.HE and PAS staining: The bronchial structure in the model group was damaged compared with the normal group,with obvious smooth muscle hyperplasia and inflammatory cell infiltration,and a large number of goblet cells and mucus substances were visible;compared with the model group,the pathological conditions of each treatment group showed different degrees of improvement.3.ELISA method: The expression of IL-13 and IL-25 in BALF of rats in the model group were significantly different(P<0.01)compared with the normal group;expression of IL-13 and IL-25 in Pingchuanning high,middle,low dose group were significantly different(P < 0.01)compared with the model group,and there was no difference in IL-13 between Guilongkechuanning and dexamethasone group,but there were differences in IL-25(P<0.01 or P<0.05).4.RT-qPCR method: Compared with the normal group,NOX4,RIG-Ⅰ and Col6 mRNA expression in the lung tissue of the model group was significantly different(P<0.01);each treatment group had statistical differences(P<0.01)compared with the model group.5.Western blot method: Compared with the normal group,NOX4,RIG-Ⅰ and Col6 protein expression in the model group were different(P<0.01);there was no statistical difference in the expression of Col6 protein in the Pingchuanning low-dose group,and for other treatment groups,there were statistical differences(P<0.01)when compared with the model group.Conclusion: In the cold asthma rats model,Pingchuanning can alleviate airway inflammation,its mechanism may be related to PI3 K regulation of ROS,then regulating RIDD-RIG-Ⅰ-NF-κB related NOX4,RIG-Ⅰ and Col6,which reduces the expression of IL-13,IL-25 and other cytokines,and interferes with inflammatory reaction. |
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