Expression And Significance Of ROR?t And SOCS1/3 In Lung Tissue Of Asthmatic Mice

BackgroundRespiratory Syncytial Virus?RSV?is a kind of RNA virus,one of Paramyxivirinae,which has 2 major serotypes and 9 subtypes,encoding 11 kinds protein including adsorptional protein G,fusional protein F,Small hydrophobic protein SH,non-structural protein NS1 and NS2.RSV always found in neonatal and 6 month old baby,which is the most common pathogens for respiratory tract infections in infants,especially the occurrence of severe bronchiolitis and pneumonia in infants 2-6 months,which is the important risk factors of bronchial asthma in children.The mechanism of RSV inducing acute attack of asthma is not clear.Most attention has been paid to the change of th1/th2 and imbalance of th17/treg,and the transcription factor retinoic acid receptor-related orphan receptor?t?ROR?t?and suppressor of cytokine signaling1/3?SOCS1/3?.ROR?t is a member of retinoic-related orphan receptor subfamily?ROR?.The specific expression of ROR?in immune cell subtypes,is the key transcription factor of human and mouse T helper 17 cells?Th17?,which can regulate the differentiation of Th17 cells and the secretion and expression of specific effects of interleukin17?interleukin 17,IL-17 role?.The main effect factor of Th17 cells is closely related to the occurrence and development of IL-17,both with many autoimmune and inflammatory diseases,and the relationship between asthma and RSV infection has been confirmed.Th17cells expressing and secreting IL-17 is a proinflammatory cytokine powerful neutrophil recruitment,which can promote the secretion of the role of inflammatory factors in many cells,resulting in airway mucus secretion,high reactivity,resulting in airway remodeling and leading to asthma.Members of the protein subfamily suppressor of cytokine signaling?SOCS?have also been shown to be associated with the development and progression of many autoimmune diseases,autoimmune diseases and inflammatory diseases.While SOCS1 and SOCS3 as an important member among them,studies have pointed out that the two and IL-6,IL-10,IL-12,IFN,influence the expression level of LPS and other inflammatory and anti-inflammatory cytokines,also can inhibit the signal transduction of immune molecules.SOCS1 and Janus kinases?JAKs?in Jak1,JAK2he and JAK3 combination,inhibit the activity of JAKs and signal transducer and activator of transcription?STAT?signaling pathway is an important function of interferon activation plays an important role in host innate antiviral responses in the cytokine signaling pathway.SOCS3 is the key factor regulating effect antiviral and immune mediated interferon can be combined with the specific cytokine receptors,inhibition of the JAK/STAT signaling pathway.RSV virus and antiviral response to interferon mediated by leading the expression level of SOCS1/3 protein affects the inhibition effect of both is also considered an important factor in anti RSV infection and interferon induced acute exacerbation of asthma in.So in order to clear ROR?t and SOCS1/3 on the expression of RSV infection in acute exacerbations of asthma have influence,this paper use the OVA induced asthma of the BALB/c mice.Repeated RSV infection was performed to observe the airway inflammation and the incidence of asthma in mice,and to explore the mechanism of the two effects on the airway inflammation.ObjectiveTo study the ROR?t?retinoic acid receptor-related orphan receptor?t,ROR?t?and SOCS1/3?suppressor of cytokine signaling1/3,SOCS1/3?protein expression and significance of lung tissue in mice infected with RSV virus?respiratory syncytial,virus,RSV?,and to further explore the pathogenesis of asthma caused by RSV infection,to provide a new direction for clinical treatment.Methods6-8 weeks old male BALB/c mice?SPF 36?were selected as the research object,they were randomly divided into the blank group,the OVA sensitized group?asthma group?and the OVA/RSV?asthma combined with RSV infection group?3 groups,12 rats in each group,the asthma model were established.Mice in the group OVA/RSV were sensitized with OVA,and the next day were injected into the nasal cavity of RSV to establish and replicate the acute RSV virus infection asthma model;mice in the OVA sensitized group were treated with the same method for OVA sensitization and stimulation;mice in blank group were treated with equal dose of PBS solution instead of OVA and RSV.The general situation of the 3 groups of mice were observed;the mouse animal ventilator were used to detect airway resistance in 3groups of mice;the bronchoalveolar lavage fluid?BALF?of each group were collected,and the inflammatory cells were classified and counted by blood cell analyzer;the lung tissues of each group were fixed,dehydrated,embedded and sliced,routine hematoxylin eosin?HE?staining was used to observe the pathological changes of lung tissue pathology of mice;the levels of ROR?tmRNA,SOCS1mRNA and SOCS3mRNA in lung tissue of 3 groups of mice were detected by real-time fluorescent quantitative PCR and reverse transcription polymerase chain reaction?RT-PCR?.Results?1?The fur of the blank control group mice were always light,the reaction were sensitive,the breathing were normal;the OVA sensitized mice and OVA/RSV mice were gathered in the corner of the glass after each excitation,and the behavior were changed from the previous restlessness to the later stage.?2?The airway resistance?R_L?in the OVA sensitized group and the OVA/RSV group were significantly higher than those in the blank group,the Lung compliance(C_dyn)were significantly lower than those in the blank group?P<0.05?,and the OVA/RSV group R_L was significantly higher than that in the OVA sensitized group,Cdyn was significantly lower than that in the OVA sensitized group?P<0.05?.?3?The proportion of eosinophils and lymphocytes in the OVA group and the OVA/RSV group mice's BALF were significantly higher than those in the blank group,and the proportion of monocytes was significantly lower than that of the blank group?P<0.05?,the proportion of BALF in the mice of OVA/RSV group were significantly different from that of OVA group in comparisons between each two groups?P<0.05?.?4?No obvious inflammatory reaction in the lung tissue of the blank group mice,blood vessels and trachea were not dilated,inflammatory corpuscle were not infiltrated,alveolar wall were smooth,no exudation in alveolar space,no collagen hyperplasia around the airway,no mucus secretion in airway;collagen hyperplasia were observed in the airway of OVA sensitized mice,there were mucus secretion in the airway,goblet cell hyperplasia multiplication were visible in the interstitium and alveoli of the lung,and there were eosinophilic granulocyte,there were a large number of inflammatory cells and bronchial mucosa thickening in the bronchi and the surrounding vessels,bronchial mucosa were thickening;in group OVA/RSV,there were obvious collagen hyperplasia around the airway,and obvious mucus secretion in the airway,there are obvious inflammatory cell infiltration in the alveoli,alveolar stenosis and capillary congestion were obvious,the mice showed typical interstitial pneumonia.?5?The expression level of ROR?tmRNA in lung tissue of the OVA sensitized group and the OVA/RSV group were significantly higher than those of the blank group?P<0.05?,and the expression level of ROR?tmRNA in lung tissue of mice in OVA/RSV group were significantly higher than those in the OVA sensitized group?P<0.05?.?6?The expression levels of SOCS1 and SOCS3mRNA in lung tissue of the OVA sensitized group and the OVA/RSV group were significantly higher than those in the blank group?P<0.05?,and the expression levels of SOCS1 and SOCS3mRNA in lung tissue of mice in group OVA/RSV were significantly higher than those in the OVA sensitized group?P<0.05?.ConclusionThe level of ROR?t and SOCS1/3mRNA expression were enhanced in lung tissue of asthmatic mice and asthmatic mice with RSV infection resulting in the enhancement of lung tissue inflammation in mice,which may be an important mechanism of the occurrence of asthma.