强直性脊柱炎患者免疫球蛋白与自噬基因和通路的变化及相关性分析

Changes and correlation of immunoglobulin and autophagy genes and pathways in patients with ankylosing spondylitis

目的 分析来自安徽省中医院风湿科的39例强直性脊柱炎(AS)患者和31例健康体检者的血清免疫球蛋白(IgG1、IgG2、IgG3、IgG4、IgA、SIgA、IgM)、淋巴细胞自噬蛋白Beclin1、自噬指标微管相关蛋白Ⅰ轻链(LC3-Ⅱ)、磷脂酰肌醇三磷酸激酶(PI3K)、AKT、哺乳动物雷帕霉素靶蛋白(mTOR)、血清自噬相关基因(Atg1、Atg5、Atg12、Atg13、Atg17),观察上述指标的变化并对其相关因素进行分析。方法 将39例AS患者设为实验组,对照组选同时期的31例正常健康人,采用ELISA方法检测两组人员的血清免疫球蛋白(IgG1、IgG2、IgG3、IgG4、IgA、SIgA和IgM)水平,运用Western blot检测两组成员的外周血淋巴细胞自噬相关蛋白Beclin1、LC3-Ⅱ和细胞自噬通路蛋白PI3K、AKT、mTOR指标的变化,利用PCR方法检查血清自噬相关基因(Atg1、Atg5、Atg12、Atg13、Atg17),并分析各指标之间的相关性。结果 实验组血清免疫球蛋白IgG1、IgG4、IgA、SIgA较健康对照组显著升高(P<0.05或P<0.01),两组IgG2、IgG3、IgM无明显变化。实验组PI3K、mTOR较健康对照组显著升高,P<0.01。两组Beclin1、LC3-Ⅱ、AKT无明显差异。实验组ATG1、ATG12、ATG13、ATG17较健康对照组明显升高,ATG5明显降低,P<0.01。相关分析结果显示,AS患者IgG1、IgG2、IgG3、IgG4与mTOR呈正相关,SIgA与Beclin1呈正相关,IgG1、IgA、IgM与AKT呈正相关,免疫球蛋白各亚型与LC3-Ⅱ、PI3K无明显相关性。AS患者IgG1、IgG2、IgG3、IgA、SIgA、IgM与ATG17呈负相关,IgG4与ATG17呈正相关;免疫球蛋白各亚型与ATG1、ATG5、ATG12、ATG13无明显相关性。结论 AS患者存在免疫球蛋白、细胞自噬通路蛋白、细胞自噬基因指标的变化,且各项指标存在一定的相关性。AS患者体液免疫和细胞自噬都与PI3K/AKT/mTOR信号通路相关,AS患者体液免疫增强的同时,PI3K/AKT/mTOR信号通路激活,细胞自噬受到抑制,细胞自噬基因出现不同程度的异常表达。

Objective To analyze serum immunoglobulin(IgG1,IgG2,IgG3,IgG4,IgA,SIgA,and IgM),lymphocyte autophagy protein Beclin1,LC3-II,PI3K,AKT,mTOR,serum autophagy related genes(Atg1,Atg5,Atg12,Atg13,and Atg17)in 39 cases with ankylosing spondylitis and 31 healthy subjects from the Rheumatology Department of Anhui Provincial Hospital of Traditional Chinese Medicine,to observe the changes of the above indexes,and to analyze the related factors.Methods Totally 39 patients with ankylosing spondylitis were classified as the experimental group,and the other 31 healthy patients were chosen as the control group.The levels of serum immunoglobulin(IgG1,IgG2,IGG3,IgG4,IgA,SIGA,and IgM)in the two groups were detected by ELISA,and Western blot was used to detect the lymphocyte of Beclin1,LC3-II,PI3K,Akt and mTOR.PCR was used to detect the autophagy-related genes Atg1,Atg5,Atg12,Atg13,and Atg17 in serum,and the correlation of the indicators was analyzed.Results Compared with those in the healthy control group,IgG1,IgG4,IgA and SIgA were significantly increased in the experimental group(P<0.05 or P<0.01);there were no significant changes in IgG2,IgG3 and IgM in both groups.PI3K and mTOR in the experimental groups were significantly higher than those in the healthy control group,with statistical significance(P<0.01);there were no significant differences in Beclin1,LC3-II and AKT between the two groups.Compared with those in the healthy control group,ATG1,ATG12,ATG13 and ATG17 in the experimental groups were significantly increased,while ATG5 was significantly decreased,with statistical significance(P<0.01).Correlation analysis results showed that IgG1,IgG2,IgG3,IgG4 were positively correlated with mTOR,and SIgA was positively correlated with Beclin1 in AS patients.IgG1,IgA and IgM were positively correlated with AKT.There was no significant correlation between the subtypes of immunoglobulin and LC3-II and PI3K.In AS patients,IgG1,IgG2,IgG3,IgA,SIgA and IgM were negatively correlated with ATG17,while IgG4 was positively correlated with ATG17.There was no significant correlation between subtypes of immunoglobulin and ATG1,ATG5,ATG12,and ATG13.Conclusion There are changes of immunoglobulin,autophagy protein and autophagy gene in patients with AS,and there are some correlations among these indexes.Both humoral immunity and autophagy in AS patients are related to the PI3K/AKT/mTOR signaling pathway.While humoral immunity is enhanced in AS patients,the PI3K/Akt/mTOR signaling pathway is activated,autophagy is inhibited,and autophagy genes are abnormally expressed to varying degrees.