Expression Of Notch Pathway Protein, Th17 And Treg Cells In Skin Lesions Of Alopecia Areata And Their Correlation

Background:Alopecia areata is caused by a variety of factors,including genetic predisposition,immune abnormalities,psychological stress,and abnormal levels of some nutrients,etc.Of all the the possible facters,follicular tissue-specific autoimmune response is the most accepted mechanism,related to which a number of studies have shown the presence of lymphocytes especially CD4+T cells around the hair follicles of acute alopecia areata patients.It has been found that Notch signal meridians or Th17,Treg cell is associated with alopecia areata,but the exact link has not been clear yet,we need to study it further.Objective:Our aim was to explore whether Notch signaling pathway,Th 17 or Treg cell is abnormal in the lesion of patients with alopecia areata.And found the correlation within these abnormal indicators.Methods:Part ?:We selected 13 cases of alopecia areata patients and 12 cases of normal scalp as research objects.The PCR detection indexes were Notch 1,Notch 4,Delta 4,Jagged 1,ROR-yt and Foxp3 mRNA,and we used the 2-?ct to value the expression of mRNA.Immunohistochemical staining labeled CD3+CD4+CD8+,IL-17+,Foxp3+cells and Notch l,Notch4,Jagged 1 protein,and the image Pro Plus detected the IOD of staininged image.Then we used program SPSS20.0 to do the statistical analysis;Part ?:We divided 18 C:3H/HeJ mice randomly to the model group and control group,modeling method was chronic stress stimuli,including clamping tail,wet cage,behavior constraints.Then we took the lesion area of modeled mice and the normal skin of controlled mice in the same part of skin to take PCR testing,and the test content and the way of statistical analysis was same as part I.Results:Part ?:The CD3+,CD4+ and IL-17+cells of acute and chronic A A patients are significantly increased than normal,Foxp3+cells are significantly lower than normal,and CD8+ cells are not significantly different between patients and controls,the five-labeled antibody cells were not significantly different between the acute and chronic AA patients,and was not significantly correlated with the severity of AA,neither significantly different in different parts of the hair follicle invasion(upper/lower).Jaggedl ligand protein expression was significantly lower in acute and chronic alopecia areata patients than the normal ones,showed no difference in the upper and lower hair follicle.While the expression of the mRNA between alopecia areata patients and the normal ones was not statistically differente(P>0.05).The expression of Notch4 protein in patients with acute and chronic alopecia areata showed no significant difference from normal people,but its mRNA expression in acute patients with alopecia areata was lower than that of normal people(P<0.05);The expression of Delta4 protein in patients with acute and chronic alopecia areata showed no significant difference(P>0.05),but the mRNA expression in acute alopecia areata patients was significantly lower than the normal ones(P<0.05).Notch1 protein and mRNA expression showed no significant difference between the acute alopecia areata patients and the normal ones(P>0.05).The abnormal expression ones,Jaggedl protein and Notch4,Delta4 mRNA,had significant correlation with the severity of alopecia areata.Then we explored the correlation between the abnormal Notch pathway proteins and the infiltration of Th17,Treg cell,the result was negative(P>0,05).Part ?:After 8 weeks,there were varying degrees of hair removal in head appeared section(including beard),abdomen,proximal extremities and the back within the model group We took the lesions from the head of model group and the head skin from the control group.The we used PCR method to detect Notch1,Notch4,Delta4,Jagged1,ROR-?t,Foxp3 mRNA expression in the lesion area,and the results showed no significant difference compared to the control group(P>0.05).Conclusion:There was a presence of lymphocytic infiltration around the hair follicles of lesion in AA patients,and the main part is CD4+T cell.The imbalance of Th17/Treg cell,which means Th17 cells increased but Treg cells decreased,in the lesions of AA patients.There was no difference in the different courses of the diseaseor different part of the hair follicle.Of the Notch signaling related proteins,Jaggedl protein may play a role in the occurrence of acute AA,Notch4 and Delta4 mRNA had some influence in the development of chronic AA.But the abnormal Notch ligands and receptors had no significant correlation with the infiltration of Th17,Treg cell.So the correlation between the pathogenesis of AA or Th17,Treg cells expression with Notch pathway remained to be further studied.In addition,chronic stimulated stress for C3H/HeJ mouse was a viable method of alopecia areata model making,and the Th 17,Treg cells,Notch signaling pathway partial protein were not involved in the stress response in these mice after the onset of alopecia areata.