| Background:Gout is a group of clinical syndromes in which monosodium urate(MSU)crystals are deposited in tissues or organs associated with tissue damaged due to elevated blood uric acid levels.Hyperuricemia is the biochemical basis of gout.One of the characteristics that distinguish gout from other inflammatory diseases is the recurrent and spontaneous remission of acute inflammation.Autophagy is a highly conserved catabolism mechanism,which mediates the degradation of damaged cells by inducing their components to fuse with lysosomes to maintain homeostasis..Autophagy is involved in the occurrence and development of gout,and the current studies mainly focus on autophagy's involvement in NLRP3 inflammas,NF-?B p62 signaling pathway and the production of inflammatory mediator il-1?.Therefore,we hope to find relevant targets that affect autophagy function and participate in gout inflammatory regulation through autophagy upstream pathway.The atgl611-atg12-atg5 complex plays an important role in the formation of autophagosomes.However,the expression of atg1611-atg12-atg5 complex in peripheral blood mononuclear cells of patients with gout and its clinical significance are still unclear.Objective:To clarify the expressions of Beclin 1,ATG16L1,ATG12,ATG5 and LC3B in patients with primary gouty arthritis(GA)at different inflammatory stages,and to preliminarily investigate the expressions and their clinical significanceof Beclin 1,ATG16L1,ATG12,ATG5 and LC3B in patients with primary gouty arthritis,providing a theoretical basis for accurate treatment of gout..Materials and methods:(1)Collect clinical data and laboratory examination indicators of 61 patients with primary gout in the outpatient and inpatient department of rheumatology and immunology in the affiliated hospital of North Sichuan Medical College from September 2018 to September 2019.There were 30 patients with acut gout(AG)and 31 patients with intercritical gout(IG)who must meet the diagnostic criteria for gout established by the American College of Rheumatology in 1977.At the same time,clinical data and laboratory examination indicators of 48 cases of health control(HC)patients who were treated in the Affiliated Hospital of North Sichuan Medical College during the same period were collected.Collect peripheral blood of three groups of study subjects and extract peripheral blood mononuclear cells(PBMCs);(2)Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect expression levels of Beclin 1,ATG16L1,ATG12,ATG5 and LC3B mRNA in PBMCs of the three groups.(3)Protein expression levels of Beclin 1,ATG16L1,ATG12,ATG5 and LC3B in PBMCs of the three groups were detected by Western Blot.(4)SPSS20.0 statistical software package and GraphPad Prism 6 were used for analysis and statistics;Mann-whitney nonparametric test was used to compare the mRNA expression differences of the above genes in the three groups;Variance analysis was used to compare the protein levels differences of the above genes in the three groups;The Spearman correlation analysis was used to analysis the correlation between the above genes and clinical indicators of gout patients.Results:1.Compared with the healthy control group,the levels of white blood cells,neutrophils,monocytes,uric acid,blood creatinine,globulin,triglycerides,cholesterol,low-density lipoprotein cholesterol,hypersensitive C-reactive protein,etc.were significantly higher in patients with gout Increased,the level of high-density lipoprotein cholesterol decreased,the incidence of hypertension and diabetes increased significantly.2.The expression level of autophagy gene transcription level in the three groups of research objects:(1)There was no statistically significant difference in the expression level of Beclinl mRNA between the three groups of research objects,0.015(0.01)vs 0.022(0.01)vs 0.020(0.01),P>0.05.(2)The expression level of ATG16L1 mRNA was significantly lower in the AG group and the IG group than in the HC group,0.0029(0.00)vs 0.0051(0.00)vs 0.0106(0.01),P was all<0.01,and there was no statistical difference between the AG and IG groups Academic significance,P>0.05.(3)The expression level of ATG12 mRNA was significantly lower in the AG group and the IG group than in the HC group,0.0186(0.01)vs 0.0206(0.02)vs 0.0336(0.04),P was all<0.05,and there was no difference between the AG and IG groups Academic significance,P>0.05.(4)The expression level of ATG5 mRNA in AG group and IG group was significantly lower than that in HC group,0.0152(0.01)vs0.0159(0.01)vs 0.0296(0.04),P were all less than 0.01,and there was no statistical difference between AG group and IG group Academic significance,P>0.05.(5)The expression level of LC3B mRNA in the AG group was lower than that in the IG group and the HC group,0.003(0.00)vs 0.003(0.00)vs 0.004(0.00),P were all less than 0.05,and there was no statistically significant difference between the IG and HC groups,P>0.05.3.The expression of autophagy gene protein level in the three groups of subjects:(1)The expression of Beclinl protein level in the AG group was lower than that in the IG group and the HC group,P<0.05,there was no statistically significant difference between the IG group and the HC group,P>0.05.(2)The expression of ATG16L1 protein level in AG and IG groups were both in HC group,P<0.05,and the expression in AG group was lower than that in IG group,P<0.05.(3)The expression of ATG12 protein level in AG group and IG group was lower than that in HC group,P<0.05,there was no significant difference between AG group and IG group,P>0.05.(4)The expression of ATG5 protein level in AG group is lower than that in IG group and HC group,P<0.05,there is no statistically significant difference between IG group and HC group,P>0.05.(5)The expression of LC3B protein level in AG group was higher than that in IG group and HC group,both P<0.05,and the expression level in IG group was higher than that in HC group,P<0.05.4.Correlation analysis of autophagy genes and clinical indicators of research subjects:(1)The expression level of Beclin 1 is negatively correlated with white blood cells and neutrophils,and positively correlated with lymphocytes;(2)ATG16L1 is associated with white blood cells,blood uric acid,blood creatinine,globulin,triglyceride Ester,cholesterol,hypersensitive C-reactive protein and low density lipoprotein cholesterol are negatively correlated and positively correlated with high density lipoprotein cholesterol;(3)ATG12 is negatively correlated with low density lipoprotein cholesterol;(4)ATG5 is correlated with globulin and low density lipoprotein cholesterol There is a negative correlation;(5)There is no correlation between LC3B and clinical indicators.Conclusions:1.Compared with the healthy control group,patients with gout,in addition to increased inflammation indicators,are also more likely to be accompanied by hypertension,diabetes,hyperlipidemia and other diseases.2.The inhibition of autophagy function may be one of the mechanisms of acute attack of gouty arthritis,and the re-activation of autophagy function is the mechanism of spontaneous remission of acute gouty arthritis.3.Beclinl,ATG16,ATG12,and ATG5 may be degraded by autophagolysosomes through ubiquitination after completing the initiation and extension of autophagosomes,or may be the result of regulation by corresponding microRNAs.4.Beclinl,ATG16L1,ATG5 may be involved in the regulation of inflammation in the pathogenesis of AG patients,and ATG16L1 may also be involved in the regulation of lipid metabolism in AG patients. |
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