Experimental Study Of The Inguinal Subcutaneous Immunotherapy For Allergic Rhinitis In Mice And The Changes Of MicroRNA In Nasal Mucosa After This Therapy

Chater1. Experimental study on the inguinal subcutaneous immunotherapy for allergic rhinitis in miceObjective:To explore the feasibility of inguinal subcutaneous immunotherapy for allergic rhinitis(AR) in mice.Methods:36 female BALB/c mice were divided randomly into six groups (n=6 per group)including the control A, the model A, the treatment A groups, and the control B,the model B, The treatment B groups(inguinal subcutaneous immunotherapy for group A, cervical back subcutaneous immunotherapy for for group B). AR model were established with ovalbumin. At 25 to 55 days, ovalbumin immunotherapy were performed in treatment groups, once two days,15 times totally. After intranasal rechallenge was performed at 56 to 62 days the AR symptom scores were documented. The eosinophils (EOS)in the nasal mucosa were measured by chromotropic acid 2R staining. Ovalbumin-specific IgE(OVA-sIgE) in the serum and expression of interferon-y and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-y and interleukin-4 was calculated.SPSS17.0 software was used to analyze the data.Results:Before treatment,The AR symptom scores of the model and treatment groups were more than 5.After treatment, the treatment A group were less than 5.The EOS count of the control A, model A, treatment A groups and the control B, model B, treatment B groups was 0.78±0.31,21.60±2.90,10.43±2.56,0.83±0.46,22.44±3.39 23.40±4.24,respectively.The EOS count of the treatment A group was significantly lower than those in model A group (t=7.08,P<0.05).There was no significant difference between treatment B and model B group (t=0.71, P>0.05).OVA-sIgE expressed was negative in control groups and positive in other groups. The ratio of interferon-y and interleukin-4 was 10.75±3.38,10.38±3.08,3.02±0.69,2.71±0.89,2.52±0.30,5.45±1.41,respectively.The ratio in treatment A group was significantly higher than those in model A group (t=5.08 here was no significant different between treatment B and model B group P>0.05).Conclusion:Inguinal subcutaneous immunotherapy indicated a good efficacy with good feasibility, short period and putting easily into practice for AR treatment in mice, which is a novel treatment method for AR in mice.Chater2. The changes of microRNA in nasal mucosa after the specific immunotherapy for allergic rhinitis in miceObjective To explore the changes of microRNA in nasal mucosa after the specific immunotherapy(SIT) for allergic rhinitis(AR) in mice.Methods Female BALB/c mice,6-8 weeks of age, were randomly divided into control group, model group and reatment group. AR model were established by intraperitoneal injection and intranasal challenge of ovalbumin and SIT was performed by inguinal subcutaneous injections. AR symptom scores were documented. The eosinophils (EOS)in the nasal mucosa were measured. Ovalbumin-specific IgE(OVA-sIgE) in the serum and expression of interferon-y and interleukin-4 in the nasal lavage were measured by enzyme-linked immunosorbent assay meanwhile the ratio of interferon-y and interleukin-4 was calculated. The microRNA in the nasal mucosa were preliminary screened by microRNA gene microarray.Comparing with model group, the Fold changes of microRNA of the treatment group were≥2.0 and the P value<0.05. MicroRNA target genes were predicted with GeneSpring12.5 software. We took the intersection between genes in the signal pathway which associated with immune response,inflammation and target genes. The MEV.4.6.0 and Cytoscape_v2.8.2. software was applied to perform the cluster analysis and target gene regulatory networks maps.Results The model of AR in mice and its SIT is successful. Comparing with the model group, the Fold changes of 15 microRNAs, of which 9 microRNAs were up-regulated and 6 microRNAs were down-regulated, were≥2.0 in treatment group(p<0.05). Cluste analysis shown clearly that microRNAs in the treatment group and model group respectively aggregated in two branches. The 15 microRNAs had 5302 target genes, of which,451 genes were related more with SIT by the intersection. One microRNA can regulate many target genes, and one gene can also be affected by many microRNAs. Their synergistic effects may be involved in the mechanism of SIT. Conclusion The expression of microRNA is changed in nasal mucosa after SIT for AR in mice and we can presume that microRNAs were involved in the process of SIT for AR; Bioinformatics methods can diminish the scope of target genes of microRNA, which will help us studying the effect of changed microRNA on its relative target genes after SIT,and make us better understanding the mechanism of the disease and its SIT.