| The First Part Experimental Research on Intervention Effect for Rats of AR Model with the Therapy of Jianpi Yiqi Tongqiao.Objective:To observe the intervention effect of Jianpi Yiqi Tongqiao Decoction on rats of allergic rhinitis model.Studing on the inhibitory effect of Jianpi Yiqi Tongqiao Decoction on AR model rats serum IL-5、IL-1β、TNF-α.Materials and methods:96 SD rats were fed adaptively for one week,96 rats were randomly divided into two groups by random number method:Blank control group(B group,n=16)and model group(M group,n=80).Then,the rat model of allergic rhinitis was induced by intraperitoneal injection of ovalbumin and motivated by nasal instillation of ovalbumin.After the model was copied successfully,The 80 rats of the model group were randomly divided into the following five groups according to the random number method:Model control group(group MC,16 rats);Jianpiyiqi group(group MJ,16 rats);Tongqiao group(group MT,16 rats);Jianpiyiqi Tongqiao group(group MJT,16 rats);Western medicine control group(group MX,16).2 hours after the last administration,the general symptoms of rats in each group were observed.The pathological changes of nasal mucosa of rats in each group were observed by HE staining,ELISA method was used to detect the contents of IL-5,IL-1 and TNF-in serum of rats in each group.Results: 1.The general state of the rats in each group was observed,The rats in the blank control group had good activity and no obvious allergic reaction.The rats in the model control group of allergy symptoms is the most obvious,mainly symptoms are scratching the nose and face,running nose,sneezing and so one.The symptoms of the treatment groups were significantly reduced.The western medicine control group and Jianpiyiqi Tongqiao group symptoms of rats were improved significantly.2.Pathological changes of nasal mucosa in rats:In the blank control group,the nasal mucosa of the rats were arranged in order.The model control group rat nasal mucosa inflammation is more serious,numerous eosinophils infiltration,obvious blood stasis.The pathological changes of the rats in the other groups were significantly reduced,and the western medicine group was the most obvious.3.The detection results of IL-5,IL-1β and TNF-α in serum of rats in each group were detected:Compared with the blank control group,the serum IL-5,IL-1β and TNF-αlevels in the other five groups of rats were significantly increased,there were significant differences(p<0.01);Compared with the model control group,the serum IL-5,IL-1β and TNF-α levels in each treatment group were reduced to varying degrees,the differences were statistically significant(p<0.01 or 0.05);Compared with rats serum IL-5,IL-1β and TNF-α content of Jianpiyiqi group and Tongqiao group were not statistically significant;Compared with rats serum IL-5,IL-1β and TNF-α content of Jianpiyiqi Tongqiao group and Western medicine control group were not statistically significant.Conclusion: 1.Jianpiyiqi Tongqiao decoction can significantly improve the symptoms of allergic rhinitis,reduce the changes of pathological nasal mucosa rat model of allergic rhinitis,Using Jianpiyiqi or Tongqiao therapy individually has certain curative effect.But the effect of Jianpi Yiqi Tongqiao Decoction is better.2.The content of IL-5,IL-1 β and TNF-α in serum of allergic rhinitis rats were significantly increased,Jianpiyiqi Tongqiao decoction can effectively inhibit the inflammatory cytokine,Using Jianpiyiqi or Tongqiao therapy individually can inhibit the expression of IL-5,IL-1β and TNF-α in serum of rats.But the effect of Jianpi Yiqi Tongqiao Decoction is better.The second Part Experimental Study on the Effect of the Therapy of Jianpi Yiqi Tong qiao on p38 MAPK Signaling Pathway in Rats Model of ARObjective: Detecting the expression of p38 MAPK,ATF-2 and phosphorylation level in AR model rats nasal mucosa were intervented by Jianpi Yiqi Tongqiao Decoction.Researching the effect of Jianpi Yiqi Tongqiao Decoction on p38 MAPK signal transduction pathway in AR model rats.Materials and methods:80 SD rats were fed adaptively for one week,80 rats were randomly divided into two groups by random number method:Blank control group(B group,n=16)and model group(M group,n=64).Then,the rat model of allergic rhinitis was induced by intraperitoneal injection of ovalbumin and motivated by nasal instillation of ovalbumin.After the model was copied successfully,The 64 rats of the model group were randomly divided into the following four groups according to the random number method:Model control group(group MC,16 rats);Jianpiyiqi group(group MJ,16 rats);Tongqiao group(group MT,16 rats);Jianpiyiqi Tongqiao group(group MJT,16 rats).Western-blot method was used to detect nasal mucosa of the AR model rats the expression levels of p38 MAPK、ATF-2、p-p38 MAPK、p-ATF-2 protein;The expression of p38 MAPK m RNA、ATF-2m RNA in nasal mucosa of the AR model rats was detected by RT-PCR method.Results: 1.Compared with the blank control group,the expression of p38 MAPK、p-p38 MAPK、ATF-2、p-ATF-2 in the nasal mucosa of the other groups were significantly increased,there was significant difference(p<0.01);Compared with the model control group,the expression of p38 MAPK、p-p38 MAPK、ATF-2、p-ATF-2 in the mucosa of rats in each treatment group was significantly lower than that of the model control group,there was significant difference(p<0.01);No significant differences between the Jianpiyiqi group and Tongqiao group(p>0.01);Compared with Jianpiyiqi group and Tongqiao group,Jianpiyiqi Tongqiao group rats mucosa the expression of p38 MAPK,p-p38 MAPK,ATF-2,p-ATF-2 protein were decreased,there was significant difference(p<0.01).2.Compared with the blank control group,the expression of p38 MAPK m RNA 、ATF-2m RNA in the nasal mucosa of rats in the other groups were significantly increased,and the difference was statistically significant(p<0.01);Compared with the Model control group,the expression of p38 MAPK m RNA and ATF-2m RNA in the mucosa of rats in each treatment group was significantly lower,and the difference was statistically significant(p<0.01);No significant differences between the Jianpiyiqi group and Tongqiao group(p>0.01);Compared with Jianpiyiqi group and Tongqiao group,Jianpiyiqi Tongqiao group rats mucosa the expression of ATF-2 and p-ATF-2 protein were decreased,there were significant difference(p<0.01).Conclusion: 1.The expression of p38 MAPK,ATF-2 protein and gene expression in nasal mucosa of AR rats model were increased,and P38 MAPK,ATF-2 protein is activated.2.The Jianpiyiqi Tongqiao decoction can reduce the expression of p38 MAPK and ATF-2 protein and gene and inhibit phosphorylation of p38 MAPK and ATF-2.3.The Jianpiyiqi Tongqiao decoction may be through inhibiting activation of p38 MAPK signaling pathway and down-regulation of AR rats model serum inflammatory factor levels.The third Part Experimental Study on the Effect of the Therapy of Jianpi Yiqi Tongqiao on NF-κB Signaling Pathway in Rats Model of ARObjective: Detecting the expression of NF-κB p65,NF-κB p50 and phosphorylation level in AR model rats nasal mucosa were intervented by Jianpi Yiqi Tongqiao Decoction.Researching the effect of Jianpi Yiqi Tongqiao Decoction on NF-κB signal transduction pathway in AR model rats.Materials and methods: 80 SD rats were fed adaptively for one week,80 rats were randomly divided into two groups by random number method:Blank control group(B group,n=16)and model group(M group,n=64).Then,the rat model of allergic rhinitis was induced by intraperitoneal injection of ovalbumin and motivated by nasal instillation of ovalbumin.After the model was copied successfully,The 64 rats of the model group were randomly divided into the following four groups according to the random number method:Model control group(group MC,16 rats);Jianpiyiqi group(group MJ,16 rats);Tongqiao group(group MT,16 rats);Jianpiyiqi Tongqiao group(group MJT,16 rats).Western-blot method was used to detect nasal mucosa of the AR model rats the expression levels of NF-κB p65、NF-κB p50、p-NF-κB p65、p-NF-κB p50 protein;The expression of NF-κB p65、NF-κB p50 m RNA in nasal mucosa of the AR model rats was detected by RT-PCR method.The level and localization of NF-κB p65,NF-κB p50 protein expression by immunofluorescence assay in rat nasal mucosa;ELISA method was used to detect the contents of IL-6、IL-8、ICAM-1、VCAM-1 in serum of rats in each group.Results: 1.Compared with the blank control group,the expression of NF-κBp65、p-NF-κBp65、NF-κBp50、p-NF-κBp50 in the nasal mucosa of the other groups were significantly increased,there was significant difference(p<0.01);Compared with the model control group,the expression of NF-κBp65、p-NF-κBp65、NF-κBp50、p-NF-κBp50 in the mucosa of rats in each treatment group was significantly lower than that of the model control group,there was significant difference(p<0.01);No significant differences between the Jianpiyiqi group and Tongqiao group(p>0.01);Compared with Jianpiyiqi group and Tongqiao group,Jianpiyiqi Tongqiao group rats mucosa the expression of NF-κBp65、p-NF-κBp65、NF-κBp50、p-NF-κBp50 protein were decreased,there was significant difference(p<0.01).2.Compared with the blank control group,the expression of NF-κBp65 m RNA、NF-κBp50 m RNA in the nasal mucosa of rats in the other groups were significantly increased,and the difference was statistically significant(p<0.01);Compared with the Model control group,the expression of NF-κBp65 m RNA、NF-κBp50 m RNA in the mucosa of rats in each treatment group was significantly lower,and the difference was statistically significant(p<0.01);No significant differences between the Jianpiyiqi group and Tongqiao group(p>0.01);Compared with Jianpiyiqi group and Tongqiao group,Jianpiyiqi Tongqiao group rats mucosa the expression of NF-κBp65 m RNA、NF-κBp50 m RNA protein were decreased,there were significant difference(p<0.01).3.Compared with the blank control group,the content of IL-6、IL-8、ICAM-1、VCAM-1 in the serum of rats in the other groups were significantly increased,and the difference was statistically significant(p<0.01);Compared with the Model control group,the content of IL-6、IL-8、ICAM-1、VCAM-1 in the serum of rats in each treatment group was significantly lower,and the difference was statistically significant(p<0.01);No significant differences between the Jianpiyiqi group and Tongqiao group(p>0.01);Compared with Jianpiyiqi group and Tongqiao group,Jianpiyiqi Tongqiao group rats serum the expression of ATF-2 and p-ATF-2 protein were decreased,there were significant difference(p<0.01).Conclusion: 1.In the course of allergic rhinitis,the activation of NF-κB signaling pathway and may promote the synthesis and secretion of inflammatory cytokines.2.Jianpiyiqi Tongqiao Decoction on allergic rhinitis intervention effect and anti-inflammatory effect may be achieved by inhibiting NF-κB signaling pathway. |
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