| Objective:To explore the mechanism and dose-effect relationship of external Therapy of tonifing qi and activating blood(KMQD nasal spray)in improving allergic rhinitis due to qi deficiency and blood stasis syndrome(miRNA-21).Method:The model of qi deficiency and blood stasis was made by exhaustive swimming with high fat diet,and then allergic rhinitis model was made with ovalbumin.36Sprague-Dawley(SD)rats were randomly divided into blank group,normal saline model group,high and middle groups.Six rats in each group were treated with Kemin Qidan nasal spray,monomethasone furoate and normal saline for 2 weeks after treatment with electrophysiology and ELISA,hemorheology,and the model was treated with Kemin Qidan nasal spray,monomethasone furoate and normal saline after 2 weeks of treatment,with the aid of electrophysiology and hemorheology of ELISA.Observation of the effect of Keminqi Dan nasal spray on body weight,symptoms of Qi deficiency and Blood stasis,physical signs and electrocardiogram of Rats by pathological Immunohistochemistry,PCR and other techniques,pathological immunity and ELISA,PCR assay were used to analyze the mechanism and dose-effect relationship.Results:1 General observation(1)Weight After modeling: the weight of rats in each model group increased significantly compared with the blank group(P<0.01),which indicated that the method of modeling had a significant effect on the body weight,and there was no significant difference in body weight among each model group(P>0.05).After 14 days of treatment: compared with the blank group,there was no significant difference between the high-dose Chinese medicine group and the blank group(P>0.05),indicating that the body weight of the high-dose Chinese medicine group returned to the level of the blank group after 14 days of administration;There was a significant difference between the low-dose Chinese medicine group and the blank group(P<0.05),indicating that after 14 days of administration,the body weight of the middle-low dose group was similar to that of the blank group,but it did not recover to the level of the blank group.There was significant difference between the western medicine group,saline group and blank group,indicating that the body weight of the two groups changed after 14 days of administration.There was significant difference between the high-dose Chinese medicine group and the saline group.The difference was in comparison with saline group(P<0.01).It is suggested that the improvement of qi deficiency and blood stasis syndrome will affect the body weight after treatment.(3)The fitting effect of dose change of gminqi Danbi spray with body weight was poor(P>0.05),and the dose-effect relationship of R2x0.0062 was weak.The dose-effect relationship between GMS and body weight was not significant(P>0.05).2 symptoms and signs(1)Compared with normal saline group: the decrease of integral was caused by middle and high dose Chinese medicine group and western medicine group on the14 th day after administration,and there was significant difference compared with normal saline group(P<0.01),middle dose Chinese medicine group had obvious difference(P<0.05),and there was significant difference between middle dose Chinese medicine group and western medicine group(P < 0.05).The control effect of qi deficiency and blood stasis as well as allergic symptoms and signs was remarkable because of the three effects.There was no significant difference between the low-dose Chinese medicine group and the normal saline group,which indicated that the curative effect was not good.(2)Compared with the blank group: there was significant difference between the saline group and the blank group(P<0.01),which indicated that the model-making effect was better;On the 14 th day after administration,the score of allergic symptoms in the low-dose Chinese medicine group decreased,but did not return tothe level of the blank group,that is,the normal situation(P<0.05).There was no significant difference between the high-dose Chinese medicine group and the western medicine group compared with the blank group(P>0.05),which indicated that the symptom integral returned to the level of the blank group at the 14 th day after administration.In comparison with the western medicine group,the difference between the normal saline group and the western medicine group was significant(P<0.01),and the curative effect of the western medicine group was obvious after the 14 th day of administration.The difference between the low-dose traditional Chinese medicine group and the western medicine group was significantly different(P<0.05).(4)Comparision of treatment in each group: the curative effect of the two groups was equal to that of middle,low and high dose Chinese medicine group at the 14 th day after administration,the difference was obvious(P<0.01);There was no significant difference between the western medicine group and the high-dose Chinese medicine group,which proved that they had the same curative effect.2 Pathology(1)Compared with the model group: compared with the model group,the blank group and the model group showed that there was a strong difference between the blank group and the model group(P < 0.01),and there was a significant difference between the blank group and the model group.After treatment,there was significant difference in pathology between the western medicine group and the model group(P<0.05),and there was a significant difference between the high-dose Chinese medicine group and the traditional Chinese medicine group(P<0.01).Compared with the model group,the western medicine group had statistical significance(P<0.05).(2)compared with the blank group: there was significant difference(P < 0.01),there was significant difference between each treatment group(except high dose Chinese medicine group)and blank group(P<0.05).No significant difference was found between the high-dose Chinese medicine group and the blank group(P>0.05).The results showed that after the treatment of high dose of traditional Chinese medicine,the pathological manifestations were relatively good.(3)Compared with the western medicine group,there was no significant difference in pathology between the low-dose Chinese medicine group and the western medicine group(P>0.05),but there was significant difference between the high-dose Chinese medicine group and the western medicine group(P<0.05).The results showed that there was no significant difference between the pathological manifestation of the treatment with western medicine and that of the low dose of traditional Chinese medicine in the middle and low dose of traditional Chinese medicine.(4)The dose-effect relationship between the dose change of Chinese medicine and the pathological score showed that the pathological integral decreased gradually with the increase of dose within the dose range.The trend of dose-effect relationship between dose change and eosinophil count showed that the fitting effect was poor and the dose-effect relationship was weak.3 Immunohistochemistry Compared with the blank group,the content of ECP protein in the AR rats and the nasal mucosa of the normal saline group showed a significant recovery,and there was a significant difference in the statistical significance(P<0.01),which indicated that the effect of AR model was obvious.Compared with the normal saline group,the content of ECP protein in the western medicine group and the high-dose traditional Chinese medicine group AR rats and the nasal mucosa tissue was significantly reduced,and there was a significant difference in the statistics(P <0.05),which indicated that the two groups had a significant back-down effect on the ECP,but did not reach the normal blank group level;There was no significant difference in the content of ECP protein in the nasal mucosa of the rest group,and the effect of the low-dose traditional Chinese medicine on ECP was not obvious.The effect of the dose change of the Chinese traditional Chinese medicine and the average optical density of the ECP was poor and the dose-effect relationship was weak.4 Cytokine detection(1)Compared with the saline group: the blank group and the saline group showed very obvious statistical difference(P<0.01),and the results were consistent in the comparison between the high-dose Chinese medicine group and the western medicine group and the normal saline group,and there was no significant difference between the control group and the normal saline group(P<0.01).The results showed that the concentration of IL-4 increased significantly(P<0.01),and decreased significantly(P < 0.01)after high-dose traditional Chinese medicine and western medicine(P<0.05).(2)Compared with the blank group: the concentration of IL-4 in the low-dose Chinese medicine group was lower than that in the high-dose Chinese medicine group,the middle-dose Chinese medicine group and the western medicine group were obvious(P<0.01),and did not reach the level of the normal blank group.However,the concentration of IL-4 in high,middle dose Chinese medicine group and western medicine group decreased obviously(P>0.05),and both reached the level of normal blank group.(3)Compared with the western medicine group: the results showed that there was no significant difference in statistics between the high and middle dose Chinese medicine group and the western medicine group(P>0.01),which indicated that the western medicine had a down-regulation effect on IL-4 in the body of AR rats,and the down-regulation effect was high.The middle dose of traditional Chinese medicine was the same,but it was significantly better than that of the low dose Chinese medicine group(P<0.01).(4)The dose-effect relationship between the dose change of traditional Chinese medicine and the dose-effect of IL-4 showed that the content of IL-4 decreased gradually with the increase of dose within the dose range.5 RT-PCR(1)Compared with the blank group: there was a significant difference between the saline group and the blank group(P<0.05),indicating that the amount of miRNA-21 was significantly decreased after successful modeling;There was a significantdifference in statistics between the low dose group and the blank group(P < 0.05),which proved that the effect after treatment was not satisfactory.There was no obvious difference between high-dose Chinese medicine,western medicine group and blank group after treatment,which indicated that both groups had a certain effect on miRNA recovery after treatment.Compared with the saline group: there was a certain difference between the western medicine group and the high-dose Chinese medicine group(P≈0.1),which indicated that there might be a comparison in the treatment between the two groups.A weak effect.(2)The dose-effect relationship between the dose change of traditional Chinese medicine and the expression of miRNA-21 showed that the fitting degree was poor and the correlation was weak.(3)MiRNA-21 and IL-4,IL-12,ECP regression analysis showed a weak correlation.6 electrophysiological detection(1)Compared with the pre-model,the ST segment of ECG in rats was significantly elevated(P<0.05),but the T wave did not change significantly(P>0.05),and the ST segment was significantly elevated(P<0.05),T wave)after modeling(P<0.05).Compared with those after treatment,there was no significant difference in ECG between the two groups(P>0.05),but there was a significant difference after modeling(P<0.05).(2)Compared with pre-model,the heart rate of rats after modeling was significantly different from that before modeling(P<0.01),but after treatment,the heart rate completely recovered to the heart rate before modeling(P<0.01).There was a significant difference in the post-R wave voltage(P<0.01),and a complete recovery to the pre-model voltage during the modeling(P<0.01),and the ventricular premature beat event occurred after the modeling,and the difference was obvious after the modeling,but no ventricular premature beat event occurred after the treatment.It could be observed that the ST segment of ECG decreased to 0.05 MV after modeling in some rats,which indicated that the effect of long-term myocardial ischemia in this rat was obvious,and there was a change of blood stasis constitution.7 HemorheologyCompared with the model group,the low shear rate,middle shear rate and erythrocyte aggregation index were significantly decreased in the treatment group(P<0.05),and the plasma viscosity in the treatment group was significantly lower than that in the model group(P<0.01).There was no significant difference in high shear index(P>0.05).Conclusion:1 The external treatment of invigorating qi and activating blood circulation(Kemin Qidan nasal spray)has a definite therapeutic effect on allergic rhinitis rats with qi deficiency and blood stasis syndrome.The curative effect of the high dose group is better than that of the western medicine group,and the curative effect is the same as that of the western medicine group.Different doses of Kemin Qidan nasal spray have different effects on miRNA-21 network.2 The mechanism of Yiqi Huoxue external treatment in the treatment of allergic rhinitis with qi deficiency and blood stasis syndrome may be to increase the content of miRNA-21,decrease the content of IL-4,then decrease the proportion of Th2,and down-regulate the release of ECP in tissues.In order to reduce the damage of nasal epithelial cells. |
