Study On The Expression Of Th17/Treg-related Cytokines In The Peripheral Blood Of Patients With Alopecia Areata

Objective: To investigate the changes in the expression of Th17/Treg-related cytokine,which is interleukin(interleukin-17,IL-17),interleukin-25(interleukin-25,IL-25),interleukin-35(interleukin-35,IL-35),interleukin-10(interleukin-10 IL-10)m RNA in the peripheral blood mononuclear cells,PBMCs(peripheral blood mononuclear cells,PBMCs)and the levels in cultured liquid in the disease of alopecia areata,and provided a basis for the treatment and prognosis of alopecia areata.Methods: Select 32 patients with alopecia areata(12 cases in the severe group,20 cases in the mild group,15 cases in the active group,17 cases in the stabilization group)as the study group,30 cases of health check-ups as the normal control group,and 32 cases of alopecia areata patients and 30 normal control groups were detected using the real-time fluorescence quantitative reverse transcription PCR(q RT-PCR)method IL-17 A,IL-25,IL-35(EBI3,p35),IL-10 m RNA expression in PBMCs.At the same time,the content of IL-17,IL-25,IL-35,and IL-10 in PBMCs cultured liquid was determined by enzyme linked immunosorbent assay(ELISA).Results: 1.The IL-17A、IL-25、IL-10、IL-35 Gene expression level in PBMCs of alopecia areata patients:(1)IL-17A、IL-25、IL-10、IL-35(EBI3、p35)m RNA relative expression: Comparisons the different were made between alopecia areata severe groups,mild groups,and normal control groups.There are statistics significance.[Z=8.315;Z=14.495;Z=10.955;Z=(13.096、11.765),P<0.05].Among them,IL-17A、IL-10 in alopecia areata severe groups were lower than normal control group and alopecia areata mild groups,IL-10 in alopecia areata mild groups was also higher than normal control group,IL-25 in the alopecia areata severe groups and mild group was lower than the normal control group.IL-35(EBI3)in the alopecia areata severe groups was lower than the normal control group and the alopecia areata mild groups,IL-35(p35)in the alopecia areata mild groups was higher than the normal control group and the alopecia areata severe groups.The comparative differences between the two groups are statistically significant(P < 0.05),the comparative differences between the remaining two groups are not statistically significant(P > 0.05).The differences in the relative expression of IL-25 、 IL-35(EBI3)m RNA between the alopecia areata active group,the stabilization period group,and the normal control group.There are statistics significance(Z=12.898;Z=10.939,P<0.05),IL-17A、IL-10 、 IL-35(p35)m RNA Relative expressions were between the alopecia areata active group,the stabilization period group,and the normal control group.There is no difference statistics significance(P > 0.05).Among them,IL-25 in the group of alopecia areata active period and the stable period group were lower than the normal control group.IL-35(EBI3)in the group of alopecia areata active periods was lower than the normal control group.The comparative differences between the two groups are statistically significant(P < 0.05),the comparative differences between the remaining two groups are not statistically significant(P>0.05).(2)Rank correlation analysis:IL-17A、IL-10、IL-35(EBI3、p35)m RNA relative expression and SALT score[r=-0.420;r=-0.447;r=(-0.420 、-0.426),P < 0.05] was negative correlation,IL-25 m RNA relative expression and SALT There was no significant correlation between the ratings(P > 0.05),IL-17,IL-25,IL-10.There was no significant correlation between IL-35 and disease activity(P>0.05).2.Alopecia areata patients PBMCs cultured supernatant fluid IL-17、IL-25、IL-10、IL-35 level:(1)The difference of IL-25 between alopecia areata severe groups,mild groups,and normal control group,there are statistics significance(Z=9.851,P<0.05),the IL-17、IL-10、IL-35 between alopecia areata severe groups,mild groups,and normal control groups.There are no differences statistics significance(P>0.05)among them,the IL-25 in alopecia areata severe group and mild group was also higher than normal control group,The comparative differences between the two groups are statistically significant(P <0.05),The comparative differences between the remaining two groups are no statistically significant(P>0.05).The difference of IL-17、IL-25 between the alopecia areata active group,the stabilization period group,and the normal control group,There are statistics significance(Z=8.894;Z=9.446,P<0.05),The difference of IL-10、IL-35 between the alopecia areata active group,the stabilization period group,and the normal control group,There are no statistics significance(P>0.05),Among them,the IL-17 in the alopecia areata stabilization group was higher than the normal control group and the alopecia areata activity group.IL-25 in the alopecia areata active group and the stable period group were higher than the normal control group.The comparative differences between the two groups are statistically significant(P<0.05),the comparative differences between the remaining two groups are not statistically significant(P>0.05).(2)Rank correlation analysis:the level of IL-17 was positively correlated with disease activity(r=0.479,P<0.05)and had no significant correlation with the salt score(P > 0.05).There was no significant correlation between IL-25,IL-10,IL-35 levels and SALT scores and disease activity in supernatant fluid(P>0.05).In the supernatant fluid IL-25、IL-10、IL-35 level with SALT,There was no significant correlation between scoring and disease activity(P>0.05).3.IL-17A、IL-25、IL-10、IL-35 correlation in patients with alopecia areata:Relative expression of IL-17 A and IL-25、IL-10、IL-35(EBI3、p35)m RNA with alopecia areata patients PBMCs is positively correlated[r=0.474;r=0.449;r=(0.567、0.562),P<0.01],IL-10 and IL-25 m RNA relative expression(r=0.501,P<0.01)is positively correlated.There was no significant correlation between the levels of IL-10,IL-17 A,IL-25,and IL-35 in PBMCs cultured serum in patients with alopecia areata(P>0.05).Conclusion: 1.Th17/Treg cytokines(IL-17,IL-25,IL-10,IL-35)are involved in the onset of alopecia areata,which is correlated with the severity of alopecia areata disease,and there is synergy between cytokines,which can provide theoretical basis for alopecia areata treatment to a certain extent.2.IL-17 and IL-25 with high levels of supernatant fluid expression in PBMCs in patients with alopecia areata are consistent,closely related to the occurrence of alopecia areata disease,and to some extent promote inflammation of alopecia areata.