The Study Of The Relationship Between Astrocyt ES And Migraine

ObjectiveThe role of astrocytes in nervous system diseases is increasingly recognized.Astrocytes in the brain can directly regulate the function,survival and activity of nerves,regulate the concentration of ions inside and outside of neurons,and transmit the second messenger,as well as ingestion,inactivation and supply of neurotransmitters.Trigeminal nerve vascular theory is the main pathogenesis of migraine.Trigeminal ganglia consists of neurons and glial cells,however the role of astrocytes in the pathophysiology and treatment of migraine is still lack of clear characteristics.Our research attempts to find out the function and role of astrocytes on the pathogenesis of migraine,and further study the pathogenesis of migraine to provide new ideas for the research on migraine.MethodsThis study intends to induce dermal cells into fibroblasts using the skin tissue of the patient group and the healthy group,and then induce fibroblasts into i PSCs(induced pluripotent stem cells),and then induce i PSCs into astrocytes,and then use primary astrocyte culture,primary neuron culture,nitroglycerin stimulation,immunofluorescence,cell transfection and q-PCR,to explore the role of astrocytes in the pathogenesis of migraine from the molecular,cellular and global levels by comparing the changes of inflammatory cytokines’ expression such as 5-HT(5-hydroxytryptophan),S100Β,NGF(nerve growth factor),CGRP(calcitonin gene-related peptide)and IL-1β(interleukin-1β)in astrocytes of migraine patients.Cytokine expression levels changed before and after nitroglycerin stimulation.Meanwhile,the effect of astrocytes on primary neurons in migraine patients was studied.This study explores the role of various inflammatory factors in the pathogenesis of migraine.ResultsThere was no difference between the disease group and the normal control group in the process of fibroblast growth from epidermal cells in the first stage.In the second stage,fibroblast induced differentiation into i PSCs,showed no difference between the disease group and the normal control group.In the third stage,i PSCs were more easily differentiated in the disease group than in the normal group during the passage and amplification of i PSCs.In the fourth stage,during the induction of i PSCs into astrocytes,it was more difficult for the disease group to form a clone sphere than the normal group,and the morphology of astrocytes in the disease group was different from that in the normal control group.The fifth stage simulated an in vitro migraine stimulation model,and the disease group and the normal group were designed with a GTN stimulation group and a non-stimulation group.There was no significant difference in the expression of S100Β,5-HT,IL,CGRP between the disease group and the normal group in the absence of nitroglycerin(-GTN)stimulation.The expression of 5-HT and IL-1β in +GTN increased compared with that in-GTN,while the expression of NGF and CGRP decreased in+GTN.and the IL-1β in neurons were significantly increased,the CGRP in neurons were significantly decreased.ConclusionsThe observation and analysis of the whole induction process showed that compared with the healthy control group,the astrocytes in the disease group were more difficult to form clonal spheres after resuspension,and the aggregation characteristics of astrocytes were reduced.The specific reasons for this need further study.IPSCs in the disease group were more easily differentiated than that in the normal group,and in the process of inducing astrocytes,the morphology of immature astrocytes in the disease group was significantly different from that in the healthy control group,which may be related to the pathophysiological mechanism of the disease itself.Biochemical indicators of migraine in the disease group and the healthy control group were measured by q-PCR and it was found that in the absence of nitroglycerin stimulation,compared with healthy control group,NGF expression was decreased in disease group.After nitroglycerin treatment,the expressions of 5-HT and IL-1β in +GTN were increased compared with that in-GTN,and the expressions of NGF and CGRP were decreased.Nitroglycerin stimulated the astrocytes to produce an inflammatory response.s The significant increase of IL-1β in neurons suggested neuroinflammation in migraine patients.SignificanceAstrocyte is important for the study of nervous system diseases.However,human astrocytes are difficult to obtain,and there are various ethical problems.Although fetal nerve cells from different region is very useful in culturing nerve cells,astrocytes differenttiated from induced pluripotent stem cells(i PSCs)are the most convenient choice to obtain disease specific organization materials.The differentiation of astrocytes has been confirmed from human ESCS and i PSCs closest to human.In this study,we induced human astrocytes,and directly observed the difference of astrocytes between migraine patients and the normal control group during the whole induction process.Then we compared the expression level of inflammatory cytokines in astrocytes between two groups to study the effect of astrocytes on primary neurons in migraine patients and investigate the role of changes of inflammatory cytokines’ expression in astrocytes in the pathogenesis of migraine.