Effect Of Seasonal Variation On The Immunologicalregulatory Mechanism Of Respiratory System

Seasonal climate is the basic conditions of human life,its changes are most likely to lead to diseases,especially respiratory diseases(RD).After body were stimulated by seasonal variation(SV),the immune system will play the role of immune defense and immune homeostasis.The partly immune balance of the affected lungs will fluctuated by SV,which will lead to RD if beyond physiological range.The immunity poikilostasis of respiratory system(RS) was a delicate process strictly regulated.The RS mechanism of inherent and adaptive immunity ensured the conduct of normal physiological function of RS by against the common attacks of adverse factors.Once this seasonal physiological and pathological transform rule were discove,the great contribution will be appeared in prevention and cure of RS. Therefore, understanding the respiratory immune homeostasis and its regulatory mechanisms by SV would had great significance if improving the level of diagnosis,treatment and prognosis on respiratory systemic diseases involved pulmonary infection.In the RS function of the immune defense,the immune related substances of inherent immune defense,such as natural killer lymphocytes,interferon,and cytokine;adaptive immune defense,such as T lymphocytes and other immunity molecule related RS defense,such as pulmonary surfactant(PS) had became a research focus.The theory of human-environment Inter Relation(HEIR) is one of the most important content of TCM,which consider that the inner lives of people recognize activities and the natural environment,social environment exists between interrelated,mutual influening complex relationship,the body occurs naturally physiological or pathologica reflect with a corresponding change.As soon as nerve-endocrine-immune regulation theory been discovered,pepoles realized that they have many similaties,particularly in the medical understanding of the lung organs function and West respiratory system,and the Wei-qi function and immunity defense function so it provide us a good link and bridges to research.At present the scientific HEIR theory had been confirmed by clinical practice,especially in lung and seasonal climate pathogenesis,but how SV affects the body's RS immune function and its material basis is not yet clear.In recent years,the scientific connotation of HEIR theory was discussed from various angles,particularly with the development of modern immunology, people found the resemblance between these TCM theory and modern medical investigation from immunology.A number of advancement had been made in SV on immune function and mechanism of the lungs,but the specific mechanism remains unclear.Based on the theory of HEIR and Fei-- Wei,this subject pose the hypothesis of SV on the RS immune substances,verify the changes and immune substances closely related activities,provided experiment base for the clinically common high seasonality prevalence of RS,meanwhile for preclinical and physical fitness medicine.The content of this study were divided into three parts.Part I:Effect of seasonal variation on expression of interleukin-1?,2,6,10 and surfactant associated protein A in healthy rat lung tissuesObjective:By observing the expression changes of IL-1?,2,6,10and SP-A of male rats in each season,to explore the effects of seasonal changes on the immunity of lung of normal rats,and to provide new ideas for the experimental basis to the cognition of pathology and physiology mechanism for seasonal attack of RS diseases.Methods: Sixty-four SD male rats, weighted 180-220g, purchased from forteen days before each solar term and divided into groups in chronological order,were randomly divided into eight groups(n=8):Vernal Begins(VB) groups,Vernal Equinox(VE) group,Summer Begins(SB) group,Summer Solstice(SS) group,Autumn Begins(AB) group,Autumn Equinox(AE) group,Winter Begins(WB) group and Winter Solstice(WS) group.All rats were housed with normal forage and drank freely under room temperature,receiving natural light. Then the rats were sacrificed by decapitation before 18 o'clock at corresponding solar term,then the IL-1?,2,6,10 and SP-A mRNA expression changes of the lung tissue in rats from each group were detected by RT-PCR method.Results 1 Effect of seasonal variation on expression of interleukin-1? in healthy rat lung tissuesThe immune materials of lung in different group were higher in spring and lower in other seasons.The groups that SB>VE>SS>VB>WB>WS>AB>AE changes in the trend.The expression of IL-1? in VE,SB and SS group was significantly higher than others groups(P<0.05 or P<0.01).There was no significant difference betwee the all spring and winter groups(P>0.05).2 Effect of seasonal variation on expression of interleukin-2 in healthy rat lung tissuesThe immune materials of lung in different group were higher in spring and lower in other seasons.The groups that VE>SS>SB>VB>WS>AB>AE>WB changes in the trend.The expression of IL-1? in VE,SB and SS group was significantly higher than others groups(P<0.01).There was no significant difference between the all spring and winter groups(P>0.05).3 Effect of seasonal variation on expression of interleukin-6 in healthy rat lung tissuesThe immune materials of lung in different group were higher in summer and lower in winter.The groups that SS>SB>VB>AB>AE>VE>WB>WS changes in the trend. Compared with WB and WS group,the expression of IL-6 was significantly higher in VB,VE,SB,SS and AB group(P<0.05 or P< 0.01);Compared with SS group,it was significantly lower in VE,AE,WB and WS group(P<0.05 or P<0.01);Compared with AB group,it was significantly lower in AE,WB and WS group(P<0.05 or P<0.01).4 Effect of seasonal variation on expression of interleukin-10 in healthy rat lung tissues The immune materials of lung in different group were higher in summer /autumu and lower in spring/winter.The groups that SB>VE>SS> VB>WB >WS>AB>AE changes in the trend. Compared with SS group,the expression of IL-10 was significantly lower in VB and WS groups(P<0.01). 5 Effect of seasonal variation on expression of SP-A in healthy rat lung tissues The immune materials of lung in different group were higher in spring /summer and lower in autumn/winter.The groups that SS>VE >SB>VB >AB >VE>WS>WB changes in the trend.Compared with VE and SS group,the expression of IL-10 was significantly lower in AB,AE,WB and WS groups (P<0.05 or P<0.01);compared with WB group,it was significantly higher in other groups(P<0.05 or P<0.01).Conclusion:The expression of IL-1 ?,2,6,10 and SP-A were adjusted by SV, which effected the immune function of lung.The immune materials of IL-1 ?,2,6,10 and SP-A maybe the mesomerism material foundation of the the immune function.The mmune function of lung existed the seasonal rhythm of lower in autumn and winterr and higher in spring and summer. It may be one of the pathophysiologic basis of respiratory diseases mostly happened in autumn and winter.Part II:Effects of SV on regulatory role of Th1 and Th2 cytokines and lung morphological changes of the SD ratsObjective:On the basis of establishing the rat model in the natural environment, we designed to observe the climatic conditions in different seasons of IL-10,INF-γin SD rat serum and lung histomorphology,expected to explain influencing the Th1/Th2 immune balance in the RS and its mechanism for guiding clinical practice and providing a theoretical basis. Methods:Animal grouping,making model and the statistical methods of data are same as part I.Morphological observation:after sacrificed by decapitation,the middle lobe of the right lung was taken and fixed with 4% paraformaldehyde and gave the dehydration,embedding and sections,then,the hematoxylin-eosin HE staining.Testing methods of INF-γand IL-10:the prepared serum was given the centrifugation,after that,the supernatants was extracted and gave subpackage.IFN-γlevel were measured with the double -antibody sandwich ABC-ELISA method,and IL-10 Radioimmunoassay.Results1 Effects of SV on lung morphological of the SD ratsSping/summer/autumn groups:there was no obvious inflammatory cell infiltration from the bronchus of lung tissues and surroundings in rats with intact wall,there were no congestion and edema in the tissue mucosa,and the alveolarcavity was clear and spacious. Winter groups:The alveolar lumen narrowed with the widening of alveolar septum,there were no the congestion and edema of tissue.2 Effects of SV on the content of Th1 cytokines of the SD ratsThe immune materials IFN-γof rat serum were lower in winter and higher in other groups.The groups that SB>VE>VB>AB>AE>SS>WB>WS changes in the trend.Compared with WB and WS groups, it was significantly higher in the other groups(P<0.05 or P<0.01);Compared with SB group,it was significantly lower in SS,AB,AE,WB and WS groups(P<0.05 or P<0.01); Compared with VE group,it was significantly lower in SS,VE,WB ana WS groups(P<0.05 or P<0.01).3 Effects of SV on the content of Th2 cytokines of the SD ratsThe immune materials IL-10 of rat serum were higher in summer and lower in other groups.The groups that SB>SS>WB>VB>WS>AB>VE>AE changes in the trend.Compared with AE group,the level of IL-10 was significantly higher in other groups(P< 0.05 or P<0.01). There was no significant difference between Spring and summer and winter groups(P>0.05). Conclusion:The content of IL-10 and IFN-γin rat's blood serum occurred seasonal change,which were adjusted by SV to effect the Th1/Th2 immunologic balance.Winter could depress the content of IFN-γand heighten IL-10 to degrade the immune function of RS,which resulted in anumber RD.Part III:Influence of SV on the ratio of CD3+,CD4+,CD8+,CD56+ in Healthy IndividualsObjective:To observed one-year CD3+,CD4+,CD8+ and CD56+ lymphocyte in healthy individuals,and studied the effect of SV on immune function at the cellular level,in order to provide accurate and objective indicators for the HEIR theory applied to clinical prevention and treatment of RD.Methods:Twenty volunteers of Our School new-enrolled students in 2007 were chosed,men and women each 10 cases,which were confirmed by our school affiliated hospital.Those was healthy,free of all infection,acute and chronic oral nasopharyngeal disease,cancer,immune disorders,or liver and kidney dysfunction,and were not used antibiotics,non-steroidal anti- inflammatory drugs and drugs affecting the immune system in the experimental period.All volunteers had full rest in a state of indoor and outdoor environment before eight experiments time,and 2mL peripheral blood collected in a quiet afternoon under fasting conditions.Lymphocytesubsets by means of flow cytometry CD3+,CD4+,CD8+ and CD56+ lymphocyte in healthy individuals was determined.Results1 Effect of SV on CD3+in Healthy IndividualsThe immune materials CD3+ of healthy human serum were higher in summer and lower in winter.The groups that changes SS>SB>AB>VB>VE >AE>WB>WS >in the trend.Compared with SB and SS group,the expression of CD3+ was significantly lower in WB and WS groups(P<0.05 or P<0.01).2 Effect of SV on CD56+in Healthy IndividualsThe immune materials CD56+ of healthy human serum were higher in summer and lower in other groups.The groups that SB>SS>VE>WB>AB>VB >AE>WS changes in the trend.There was no significant difference betwee all groups(P>0.05).3 Effect of SV on CD4+in Healthy IndividualsThe immune materials CD4+ of healthy human serum were higher in summer and lower in winter.The groups that SS>SB>VE>VB>AB>AE>WS >WB changes in the trend.Compared with SB and SS group,the expression of CD4+ was significantly lower in WB and WS groups(P<0.01);There was no significant difference betwee spring and autumu groups(P>0.05). 4 Effect of SV on CD8+in Healthy IndividualsThe immune materials CD8+ of healthy human serum were higher winter in and lower in summer.The groups that WS>AB>VB>WB>AE>VE>SB>SS changes in the trend.Compared with SS group,the expression of CD8+ was significantly higher in WS groups(P<0.05).5 Effect of SV on the ratio of CD4+/ CD8+ in Healthy IndividualsThe ratio of CD4+/ CD8+ were higher in summer and lower in winter.The groups that SS>SB>VE>VB>AB>AE>WB>WS changes in the trend. Compared with SB group,the expression of CD4+/ CD8+ was significantly lower in WS groups(P<0.05);Compared with SS group,it was significantly lower in AB,AE,WB and WS groups(P<0.05 or P<0.01). Conclusion:There were a certain accommodation role by SV on immune functions,which tendency were strong in spring/summer weak in autumn/winter,with the corresponding recognition between Chinese HEIR theory and lung health awareness defense theory.Spring and summer could adjust the number of CD3+,CD4+,CD8+ T cell in blood serum,by heightening CD4+ and degrading CD8+,thus up-regulated the ratio of CD4+/CD8+ to emerge immunological enhancement.