Gene Expression Of NPY??-EP?CGRP In PBMCs Of Alopecia Areata And The Regulation Of APS On It

Objective: Through detecting the gene expression of neuropeptide Y(NPY)??-endorphin(?-EP)? calcitionin gene-related peptide(CGRP)in peripheral blood mononuclear cells(PBMCs)of alopecia areata patients and investigating the influence affected by astragalus polysaccharides(APS)on gene expression,we intended to explore preliminarily the role of NPY??-EP?CGRP in pathogenesis of alopecia areata and the regulation of APS on them.Methods: 20 alopecia areata patients were selected as study group(including 17 cases of localized group,3 cases of extensive group,10 cases of active group,10 cases of stable group)alopecia areata patients were divided into APS group and self-controlled group.Then 20 cases of healthy physical examiners in the same period from our hospital were recruited as control group.We used Real-time polymerase chain reaction(RT-RCR)to calculate the mRNA expression levels of NPY??-EP and CGRP that were stimulated by phytohemagglutinin(PHA)in PBMCs of the alopecia areata patients and control group.We detected the change of mRNA expression of NPY ? ?-EP and CGRP in alopecia areata patients of PBMCs of alopecia areata patients stimulated by APS and PHA.Results: 1.The gene expression of NPY ? ?-EP ?CGRP stimulated by PHA in PBMCs of alopecia areata patients:(1)NPY mRNA expression level in PBMCs of alopecia areata patients: the relative expression of NPY mRNA in the control group was(-3.10±0.31),the localized alopecia areata group was(-4.88±0.75),the extensive alopecia areata group was(-4.82±1.04),the difference among the three groups was statistically significant(F=45.353,P<0.01).Thereinto,the relative expression of the localized alopecia areata group was obviously lower than the control group,the difference between those two groups was statistically significant(P<0.01).Also,the relative expression of extensive alopecia areata patients group was lower than control group,and the relative expression of localized alopecia areata patients group was lower than the extensive alopecia areata patients group,among which there was no statistically significant differencs(P>0.05).The relative expression of NPY mRNA in the active alopecia areata patients group was(-4.95±0.84),in the stable alopecia areata patients group was(-4.83±0.72),and the difference among the three groups was statistically significant(F=45.885,P<0.01).In addition,the relative expression of active alopecia areata patients group and the stable alopecia areata patients group were both lower than the control group,the difference among those groups was statistically significant(P<0.01),the relative expression of active alopecia areata patients group was lower than the stable alopecia areata patients group,there was no statistically significant difference between them(P>0.05).(2)?-EP mRNA expression level in PBMCs of alopecia areata patients:the relative expression of ?-EP mRNA in the control group was(-4.91±0.68),the localized alopecia areata patients group was(-5.21±0.79),the extensive alopecia areata patients group was(-5.36±0.85),there was no statistically significant differencse anong the three groups(F=0.986,P>0.05).Thereinto,the relative expression of ?-EP mRNA in the active alopecia areata patients group was(-5.60±0.49),in the stable alopecia areata patients group was(-4.86±0.86),the difference among the three groups was statistically significant(F=3.978,P<0.05).Also,the relative expression of the active alopecia areata patients group was lower than the stable alopecia areata patients group and the control group,the difference among the groups was statistically significant(P<0.05),whereas the relative expression of the stable alopecia areata patients group was higher than the control group,there was no statistically significant differencse between them(P >0.05).(3)CGRP mRNA expression level in PBMCs of alopecia areata patients patients: the relative expression of CGRP mRNA in the control group was(-3.81±0.39),the localized alopecia areata patients group was(-4.11±0.53),the extensive alopecia areata patients group was(-4.52±0.60),the difference among the three groups was statistically significant(F=3.914,P<0.05).The relative expression of the extensive alopecia areata patients group was lower than the control group,and the difference between them was statistically significant(P<0.05).The relative expression of extensive alopecia areata patients group was lower than the localized alopecia areatapatients group,and the relative localized alopecia areata patients was lower than the control group,but there was no statistically significant differencse anong the groups(P>0.05).Besides,the relative expression of CGRP mRNA in the active alopecia areata patients group was(-4.44±0.48),in the stable alopecia areata patients group was(-3.91±0.50),the difference among the three groups was statistically significant(F=6.869,P<0.05).Thereinto,the relative expression in the active alopecia areata patients group was lower than the stable alopecia areata patients group and the control group,the difference among the groups was statistically significant(P<0.05),but the relative expression in stable alopecia areata patients group was lower than the control group,there was no statistically significant differencse between them(P>0.05).2.The gene expression of NPY??-EP?CGRP was stimulated by APS and PHA in PBMCs of alopecia areata patients:(1)NPY mRNA expression level in PBMCs of alopecia areata patients:the relative expression of NPY mRNA in the localized APS group was(-4.83±0.73),in the self-controlled group was(-4.88±0.75),in the extensive APS group was(-4.15±0.96),in the self-controlled group was(-4.82±1.04),in the active APS group was(-4.89±0.72),in the self-controlled group was(-4.95±0.84),in the stable APS group was(-4.57±0.84),in the self-controlled group was(-4.83±0.72).Thereinto,the relative expression of NPY mRNA in the localized APS group?the extensive APS group ? the active APS group ? the stable APS group both were higher than the self-controlled group,but there was no statistically significant differencse anong the groups(P>0.05).(2)?-EP mRNA expression level in PBMCs of alopecia areata patients patients:the relative expression of ?-EP mRNA in the localized APS group was(-5.23±0.70),in the self-controlled group was(-5.21±0.79),in the extensive APS group was(-4.89±0.69),in the self-controlled group was(-5.36±0.85),in the active APS group was(-5.59±0.49),in the self-controlled group was(-5.60±0.49),in the stable APS group was(-4.76±0.63),in the self-controlled group was(-4.86±0.86).The relative expression in the localized APS group was lower than the self-controlled group,however the relative expression in the extensive APS group?the active APS group?the stable APS group both were higher than the self-controlled group and there was nostatistically significant differencse anong the groups(P>0.05).(3)CGRP mRNA expression level in PBMCs of alopecia areata patients patients:the relative expression of CGRP mRNA in the localized APS group was(-4.21±0.42),in the self-controlled group wad(-4.11±0.53),in the extensive APS group was(-3.92±0.45),in the self-controlled group was(-4.52±0.60),in the active APS group was(-4.40±0.22),in the self-controlled group was(-4.44±0.48),in the stable APS group was(-3.93±0.46),in the self-controlled group was(-3.91±0.50).The relative expression in the localized APS group and the stable APS group were lower than the self-controlled group,the extensive APS group and the active APS group both were higher than the self-controlled group,but there was no statistically significant differencse anong the groups(P>0.05).Conclusion: 1.Our analyses showed a decreased of mRNA expression levels of NPY??-EP and CGRP in PBMCs from alopecia areata patients compared with control group and suggested that the three neuropeptides might be closely involved in the pathogenesis of alopecia areata patients.2.Stressful events might affect the release of NPY??-EP and CGRP that belonged to the neuropeptides interat closely with peripheral blood immunocytes.The down-regulated expression of NPY ? ?-EP ? CGRP in PBMCs of alopecia areata patients suggested that neuroimmune factors might play a role in the pathogenesis of of alopecia areata patients.3.APS showed no apparent regulating effect on NPY??-EP?CGRP in PBMCs of alopecia areata patients.