ABSTRACT
Objective:To investigate the changes of T-lymphocyte subsets, T-cell immunoglobulin and mucin domain molecule-1 (TIM-1) and TIM-3 levels, and cytokines in the peripheral blood of patients with active tuberculosis.Methods:From December 2017 to December 2018, 50 tuberculosis patients and 50 cured tuberculosis patients in Zhejiang Hospital of Integrated Chinese and Western Medicine were selected as the tuberculosis group and cured tuberculosis group, respectively. Fifty healthy individuals in the same period were selected as the control group. Flow cytometry was used to detect the T-lymphocyte subsets in the peripheral blood. The mRNA levels of TIM-1, TIM-3, interferon(IFN)-γ and interleukin(IL)-4 in peripheral blood mononuclear cells (PBMC) were detected by quantitative real-time polymerase chain reacticn (PCR). T test was used for statistical analysis. Results:The ratio of CD4 + /CD8 + T lymphocytes in the tuberculosis group (1.21±0.50) decreased significantly, comparing with those in the cured tuberculosis group (1.88±0.62) and the control group (1.92±0.82). The differences were statistically significant ( t=2.148 and 2.207, respectively, both P<0.05). The mRNA levels of TIM-1, TIM-3 and IL-4 in PBMC in the tuberculosis group were 2.16±0.37, 1.59±0.36 and 1.52±0.69, respectively, which were all higher than those in the cured tuberculosis group (1.60±1.23, 1.01±0.52 and 0.91±0.36, respectively) and the healthy control group (1.40±0.27, 0.92±0.34 and 0.79±0.42, respectively). All of these differences were statistically significant ( t=14.120, 11.440, 17.130, 12.090, 12.050 and 17.030, respectively, all P<0.05). However, the IFN-γ mRNA level (0.43±0.11) was lower than that in the cured tuberculosis group (1.74±0.72) and the control group (1.82±1.17), and the differences were both statistically significant ( t=13.880 and 11.430, respectively, both P<0.05). Conclusion:The immune dysfunction in patients with active tuberculosis may be related to the low ratio of CD4 + /CD8 + T lymphocytes, the increased expressions of TIM-1 and TIM-3, and the imbalance of helper T lymphocyte (Th)1/Th2 cytokines.
ABSTRACT
Objective To analyze the expression of miRNA-29a in U937 macrophages infected with Mycobacterium tuberculosis and its regulation of target genes.Methods The target genes of miRNA-29a were predicted with softwares miRnada,RNAhybrid and targetscan.The differentiation of U937 macrophages was induced by phorbol ester(PMA), the induced U937 cells were infected with bacilli calmette guerin (BCG).The expression levels of miRNA-29a and its target genes in U937 cells were detected with real-time fluorescence quantitative PCR(RT-fqPCR).The miRNA-29a over-and low-expression U937 macrophage cell lines were constructed and the levels of miRNA-29a and its garget genes were detected.The SPSS 18.0 software was used to analyze the data.Results As predicted by relevant softwares,the miRNA-29a regulate the expression of VEGFA,NFATC3 and TSC22D3 genes.After BCG infection,the expression of miRNA-29a increased to 1.33 fold(P <0.05), and the expression levels of VEGFA, NFATC3 and TSC22D3 were increased to 5.34,99.25 and 2.12 fold,respectively(P<0.01).In the miRNA-29a over-expressing U937 macrophages,the expression levels of VEGFA,NFATC3 and TSC22D3 were up-regulated to 1.35,1.29 and 3.38 fold,respectively(P<0.05 or <0.01).While in the U937 macrophages with miRNA-29a knock-down,the expression levels of VEGFA, NFATC3 and TSC22D3 were down-regulated to 0.07, 0.08 and 0.55 fold, respectively(P <0.01).Conclusion The results suggest that Mycobacterium tuberculosis infection can increase the expression of miRNA-29a in U937 macrophages,further targeting the regulation of VEGFA, NFATC3 and TSC22D3 gene expression, which may participate in the pathogenesis and development of tuberculosis.