Objective To detect the expression of autophagy-associated genes(ATGs)involved in the early stage of autophagy in peripheral blood mononuclear cells(PBMCs)of patients with ankylosing spondylitis(AS)and analyze the results with clinical data.To explore the association between the early stage of autophagy and AS and the clinical significance of ATGs involved in the early stage of autophagy in assessing the disease and inflammatory status of AS.Methods ①Clinical data and peripheral blood samples of 90 patients with AS(AS group)who were admitted to the Rheumatology and Immunology Department of the Affiliated Hospital of North Sichuan Medical College from March 2021 to August 2022 were collected,including 32 patients in the active stage(ASA group)and 58 patients in the stable stage(ASS group).In addition,clinical data and peripheral blood samples of 30 patients who were treated with secuchiumab for 24 weeks were also collected.The clinical data and peripheral blood samples of 45 healthy control(HC group)who underwent physical examination at the Affiliated Hospital of Sichuan Northwest Medical University at the same time served as controls for clinical data and peripheral blood specimens were used as controls.RT-qPCR was used to detect the mRNA expression levels of 8 ATGs(ATG13,ATG14,ATG17,ATG18,ATG101,Beclin1,ULK1,mTOR)involved in the early stage of autophagy in all PBMCs of peripheral blood samples,and compared between different groups.Data that follows a normal distribution is tested using the t-test,while data that does not follow a normal distribution is tested using the Wilcoxon rank sum test.Correlation analysis is performed using Spearman's rank correlation coefficient.②Receiver operating curve(ROC)was used to evaluate the value of ATGs with differential expression between AS group and HC group in detecting AS disease.Results ①Compared with HC group,the level of ATG13,ATG14,ATG17,ATG18,ATG101 and Beclin1 mRNA in AS group were significantly lower than those in HC group[ATG3:3.52(1.95,5.09)×10-3,7.21(5.49,9.16)×10-3,Z=-5.64,P＜0.001;ATG14:2.48(1.85,3.64)×10-3,6.16(4.27,7.80)×10-3,Z=-6.44,P＜0.001;ATG17:6.45(3.29,9.48)×10-3,18.52(12.30,22.51)×10-3,Z=-6.18,P＜0.001;ATG18:2.97(1.77,4.37)×10-3,4.61(3.27,5.59)×10-3,Z=-3.88,P＜0.001;ATG101:2.07(1.11,3.33)×10-3,3.65(2.41,5.20)×10-3,Z=-3.87,P＜0.001;Beclin1:3.50(0.63,6.14)×10-3,4.17(2.82,7.93)×10-3,Z=-1.82,P=0.027].②Comparison between ASA and ASS groups:The levels of ATG17,ATG 101 and Beclin1 mRNA in ASA group were significantly lower than those in ASS group[ATG17:4.61(2.75,7.85)×10-3,6.86(3.85,11.28)×10-3,Z=-2.16,P=0.030;ATG 101:0.93(0.40,1.67)×10-3,2.15(1.17,3.20)×10-3,Z=-3.94,P=0.002;Beclin1:1.24(0.52,3.94)×10-3,3.86(1.55,5.45)×10-3,Z=-2.26,P=0.024].③Spearman correlation analysis showed that the mRNA expression levels of ATG13 and Beclin1 in AS were negatively correlated with ESR and hs-CRP,respectively(r=-0.22,P=0.038;r=-0.30,P=0.006;r=-0.34,P=0.004;r=-0.241,P=0.037),ATG 18 mRNA expression ESR was positively correlated(r=0.22,P=0.041).④ROC curve showed that ATG13,ATG14,and ATG17 had a good ability to predict AS,and their area under the curve(AUC)was 0.821,0.866,and 0.851,respectively.The mRNA expression levels of ATG 13,ATG 14,and ATG18 in AS were significantly increased after 24 weeks of secuchiumab treatment[ATG13:3.09(0.17,4.48)× 10-3,3.50(3.42,3.57)×10-3,Z=-3.45,P=0.001;ATG14:2.49(1.43,4.03)×10-3,5.62(2.28,6.77)×103,Z=-3.01,P=0.003;ATG 18:2.91(1.61,4.37)×10-3,4.53(2.91,5.73)×10-3,Z=-3.34,P=0.001].Conclusion ①The different expressions of ATG 13,ATG 14,ATG 17,ATG 18,ATG 101,and Beclin1 between HC and AS suggest that these 6 genes may related to the pathogenesis of AS,among which Beclin1 and ATG 13 are closely related to the levels of inflammatory indicators ESR and hs-CRP in patients.It may affect the inflammatory state in AS.②IL-17Ai may be a potential regulator of autophagy,which can play a therapeutic role by affecting the early autophagy process in AS,but the specific mechanism needs to be further explored.③ Genes related to the early stage of autophagy are expected to be biological indicators for monitoring the occurrence of AS.
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