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The Expression Of ROR?t Factor In Different Tissues Of Mice Under The Action Of Fine Particulate Matter

Posted on:2021-04-20Degree:MasterType:Thesis
Country:ChinaCandidate:X S YangFull Text:PDF
GTID:2404330605964730Subject:Physiology
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The IARC and the WHO have classified air particles as grade 1 carcinogens.Quartz dust is also one of the major occupational hazards in China.As fine particles are highly adsorbent substances,they are easy to become carriers of toxic substances in the air and enter the circulatory system through alveolar capillaries,thus causing harm to multiple systems of the whole body.Two new subtypes of CD4+T cells,Th17 and Treg,have been the focus of recent research.RORyt,a specifical subtype of RORy expressed in immune cells,is the main transcription factor of human and mouse in Thl7 cells.It can not only promote the differentiation of Th17 cells,but also regulate the expression and secretion of IL-17,a specific effector of Th1 cells.ROR? is expressed in thymus,muscle,brain,heart,kidney,liver and lung,and is involved in lymph node development,immune response and production of Th17 cell.RORyt is closely related to the occurrence and development of various immune diseases,infectious diseases and tumors.In this experiment,mice were taken as the research objects,and the low-concentration particulate matter group,the medium-concentration particulate matter group,the high-concentration particulate matter group,the low-concentration staphylococcus aureus group,the medium-concentration staphylococcus aureus group,the high-concentration staphylococcus aureus group and the normal saline group were set.The changes in the mRNA expression level and protein expression level of ROR exposing t in liver,spleen and lung tissues of the mice were detected by injecting particulate matter into the trachea;At the same time,the liver,spleen and lung tissues of mice were collected as test samples,and the pathological changes of tissues and organs in each group were observed by HE staining technique;And the effect of particulate matter on immune cells of the body was speculated by comparing mice infected with staphylococcus aureus.The main findings are as follows:1.The weight of mice in the experimental group was significantly different from that in the control group,and the higher the concentration of particulate matter was,the higher the concentration of bacterial solution was,and the slower the growth of body weight was.Mice in the medium-concentration particulate matter group and the high-concentration particulate matter group ate less food,their hair was dull and sparse,and their spirits were depressed.Mice in these groups began to have cough symptoms on day 5 after lung irrigation.2.Real-time quantitative PCR results showed:In spleen and lung,the expression level of RORyt factor was significantly related to the concentration of fine particles,and the higher the concentration of particles was,the higher the expression level of RORyt factor was.Among them,the lung,as the organ directly stimulated,significantly increased the expression level of ROR?t factor when the concentration increased from 10mg/ml to 20mg/ml(p<0.05).However.the expression of ROR?t factor in the liver was not significantly higher than that in the other two organs.When the particle concentration was 20mg/ml.the expression level of ROR?t factor in liver and lung was similar to the staphylococcus aureus group when the concentration was 2*10^1 0cfu/ml.At the particle concentration of 40mg/ml.the expression level of ROR?t factor in the spleen was similar to that at the concentration of staphylococcus aureus of 1*10^11cfu/ml.3.Western blot results showed that there was a significant relationship between the expression of ROR?t protein and the concentration of fine particles in liver and lung,and the higher the concentration of particles was.the higher the expression of ROR?t protein was.In the spleen,the expression level of ROR?t protein increased significantly when the particle concentration reached 40mg/ml(p<0.05).At the particle concentration of 40mg/ml.the expression of ROR?t protein in the lungs was similar to the staphylococcus aureus group at the concentration of 2*10^0cfu/ml.At the particle concentration of 20mg/ml.the expression level of ROR?t protein in the liver and spleen was similar to the staphylococcus aureus group at the concentration of 1*10^11 cfu/ml.4.Section results showed that lung damage was the most serious under the stimulation of fine particulate matter,and the experimental group of mice all had different degrees of bronchiectasis and alveolar damage.The spleen also produced a large number of immune cells after stimulation,while the liver showed no significant changes.In this experiment.the expression of RORyt.a cellular immune factor,and pathological changes in some organs and tissues of mice in different experimental groups were analyzed by exogenous stimulation.Provide the basis for further in-depth research for damaged to the body caused by fine particulate pollutants in the atmosphere,provide evidence for air pollution and environmental protection,provide a new way for the treatment of particulate matter pollution and a new way for drug interference.
Keywords/Search Tags:Fine particulate matter, ROR?t, Real-time quantitative PCR, Western blot, HE staining
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