| Objective To investigate the changes of Th22 cell ratio and related biological characteristics in peripheral blood of patients with type 2 diabetes mellitus(T2DM)combined with pulmonary tuberculosis(PTB).Meanwhile,the effects of serum induction and exogenous interleukin-22(IL-22)on the function of THP-1 macrophages were observed in vitro,which provided experimental reference for the clinical study of T2DM complicated with PTB.Methods A total of 113 patients were collected from Linyi People’s Hospital from May2019 to April 2020.According to clinical diagnosis,these patients were divided into three groups:36 patients with type 2 diabetes combined with Pulmonary tuberculosis(T2DM-PTB),38 patients with simple pulmonary tuberculosis(PTB),39 patients with type 2 diabetes and no obvious complications(T2DM).Meanwhile,35 people with normal physical examination were collected as the control group.The research methods were as follows:(1)Flow cytometry was used to detect the ratios of Th22 cells in the peripheral blood of four groups.(2)The expression level of IL-22 cytokine in the serum of four groups were determined by enzyme-linked immunosorbent assay.(3)The mRNA expression levels of Runx1,AHR and C-Maf were detected in Th22 cells of four groups by real-time fluorescent quantitative PCR technology.(4)The ratios of PD-1~+and CD57~+cells in Th22 cells of four groups were determined by flow cytometry.(5)The mRNA expression levels of ABCA1 andβ-defensin in THP-1 macrophages were determined by real-time fluorescent quantitative PCR technology.SPSS 22.0 was used for statistical analysis.Mean±Standard deviation(x±s)was used to represent measurement results.LSD-t test was used for statistical analysis between two pairs,and P<0.05 was statistically significant.The Ethics and Academic Committee of the hospital reviewed and approved this study.All clinical patients in the study were aware of the relevant research process and had signed an informed consent form.Results(1)Analyzing and comparing the ratios of Th22 among four groups,the T2DM-PTB group was significantly higher than the control group(P<0.001),PTB group(P<0.05)and T2DM group(P<0.001);PTB group and the T2DM group were higher than the control group(P<0.001,P<0.05);PTB group was higher than the T2DM group(P<0.001);and the differences were statistically significant.(2)Comparing the expression level of IL-22 among four groups,the T2DM-PTB group was lower than the control group(P<0.001),PTB group(P<0.001)and T2DM group(P<0.001);PTB group and T2DM group were lower than control group(P<0.001,P<0.001);PTB group was higher than T2DM group(P<0.01);and the differences were statistically significant.(3)Compared with the control group,the expression level of AHR mRNA in Th22 cells was down-regulated in T2DM-PTB group,PTB group and T2DM group(P<0.05,P<0.05,P<0.05);compared with the PTB group and T2DM group,the expression level of AHR mRNA in Th22 cells in the T2DM-PTB group was also significantly down-regulated(P<0.05,P<0.05);and the differences were statistically significant;however,there was no significant difference in AHR mRNA expression between the PTB group and the T2DM group in Th22 cells(P>0.05).(4)Compared with the control group,the expressions of C-Maf mRNA in Th22 cells in T2DM-PTB,PTB and T2DM groups were significantly up-regulated(P<0.001,P<0.05,P<0.05);compared with the PTB group and T2DM group,the expression of C-Maf mRNA in Th22 cells in the T2DM-PTB group was significantly upregulated(P<0.01,P<0.01);and the differences were statistically significant;but there were no significant differences in the expression of C-Maf mRNA in Th22 cells between PTB and T2DM groups(P>0.05).(5)The expressions of Runx1 mRNA in Th22 cells among four groups were no significant differences(P>0.05).(6)Compared with the control group,the ratios of PD-1~+and CD57~+cells in Th22 cells were significantly increased in the T2DM-PTB group,PTB group,and T2DM group(P<0.001,P<0.001,P<0.001);compared with the PTB and T2DM groups,the ratio of PD-1~+and CD57~+cells in Th22 cells were significantly increased in T2DM-PTB group(P<0.001,P<0.001);the ratio of PD-1~+and CD57~+cells in Th22 cells in T2DM group was significantly higher than that in PTB Group(P<0.001);and the differences were statistically significant.(7)The mRNA expression of ABCA1 in THP-1 macrophages induced by serum in the T2DM-PTB group was down-regulated compared to the control group,PTB group,and T2DM group(P<0.001,P<0.05,P<0.05);the mRNA expressions of ABCA1 in the PTB group and T2DM group were down-regulated compared to the control group(P<0.01,P<0.01);and the differences were statistically significant;however,there were no statistically significant difference in the expressions of ABCA1 mRNA between the PTB group and the T2DM group(P>0.05).(8)The mRNA ofβ-defensin in THP-1macrophages induced by serum in the T2DM-PTB group was down-regulated compared to the control group,PTB group,and T2DM group(P<0.001,P<0.001,P<0.01);the expression ofβ-defensin mRNA in the T2DM group was lower than that of the control group(P<0.01);and the differences were statistically significant;however,there were no statistically significant difference in the expressions ofβ-defensin mRNA between the PTB group and the T2DM group(P>0.05).(9)IL-22 was injected to the T2DM-PTB group,we found that the mRNA expressions of ABCA1 andβ-defensin in THP-1macrophages induced by serum were significantly up-regulated compared to the control group(P<0.001,P<0.001),the differences were statistically significant.Conclusion(1)The disorder of Th22 cells and its cytokine IL-22 may be closely related to the occurrence and development of T2DM with PTB disease.(2)Th22 cells in patients with type 2 diabetes complicated with pulmonary tuberculosis showed an obvious aging phenotype.Meanwhile,IL-22 cytokines dependent the transcription factors expression was down-regulated and the production of IL-22 was reduced,which ultimately led to the impairment of the efflux function and bactericidal function of macrophages. |
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