A Preliminary Study Of TRIM Protein Family In The Diagnosis And Pathogenesis Of Tuberculous Pleurisy

Mycobacterium tuberculosis is a kind of intracellular infectious bacteria.The host's innate immune response has been playing an important role in the fight against tuberculosis infection.Triartite motif(TRIM)family can participate in the innate immune response of the body as an immunomodulatory protein,and can regulate the signal pathway of innate immunity a variety of ways..Its abnormal expression is related to the pathogenesis of many diseases,such as viral infection,tumor and neurodegenerative diseases.Recent studies showed that some members of the TRIM family were differentially expressed in latent infections and active pulmonary tuberculosis,and their expressions were down-regulated in patients with active pulmonary tuberculosis.On the other hand,over-expression of some members of TRIM protein family can promote the migration and proliferation of cancer cells.Objective:To screen the members of TRIM protein family that may differentially express in tuberculous pleural effusion and malignant pleural effusion,and to verify the possibility of TRIM protein family as a diagnostic and differential diagnostic aid for tuberculous pleurisy.Meanwhile,the regulatory role of TRIM4,16,27,32,35,46,47,65 and 68 9 genes in tuberculosis infection was discussed.Methods:Based on previous studies,the expression levels of TRIM4,TRIM 16,TRIM27,TRIM32,TRIM35,TRIM46,TRIM47,TRIM65,TRIM68 in tuberculous pleural effusion and malignant pleural effusion were analyzed by real-time fluorescence quantitative polymerase chain reaction(PCR)for the first time.Subsequently,the model strain Mycobacterium smegmatis(M.sm)was used to infect THP-1 macrophage line in vitro,and the cell infection model was constructed to further verify the expression changes of these nine TRIM genes.Results:This study included 53 subjects,including 32 tuberculous pleural effusion patients and 21 malignant pleural effusion patients.The expression of TIRM16 gene in tuberculous pleural effusion was 0.0077,which was significantly lower than that in malignant pleural effusion(0.0000988),P<0.0001.There was no significant difference in the expression level of other TRIM proteins between the two groups.The receive operating characteristic curve(ROC)of the subjects was analysis of the diagnostic effect of TRIM 16 shows that it has good accuracy.The area under the curve(AUC)was 0.937.In vitro infection cell experiment,it was found that the expression levels of the four genes TRIM35,TRIM46,TRIM65 and TRIM68 showed the same trend,that is,after 30 minutes of infection with Mycobacterium smegmatis,there was a significant downward trend of these four genes.After 2 hours of infection and 3 hours of infection,there was a slight downward trend and then decline gradually,which lasted for 12 hours after infection.The expression level of the four genes TRIM35,TRIM46,TRIM65 and TRIM68 began to rise gradually after 12 hours of infection.Conclusion:The expression of TRIM16 gene in patients with tuberculous pleural effusion was significantly lower than that in patients with malignant pleural effusion.ROC curve analysis showed that TRIM16 had a good diagnostic accuracy.TRIM16 gene may have potential value in the diagnosis and differential diagnosis of tuberculous pleurisy.After Mycobacterium tuberculosis model strain Mycobacterium smegmatis was infected with macrophage cell line THP-1 in vitro,the expression of four genes,TRIM35,TRIM46,TRIM65 and TRIM68,had certain significance,which suggested that it may related to the process of tuberculosis infection.