1,25-二羟维生素D(1,25-(OH)2D)和CYP27A1 mRNA表达与肺结核的相关性研究

The Correlation Between 1,25-Dihydroxyvitamin D(1,25-(OH),D)and CYP27A1 mRNA Expression with Pulmonary Tuberculosis

目的 探讨肺结核(PTB)患者中1,25-二羟维生素D(1,25-(OH)_(2)D)水平和CYP27A1 mRNA表达水平的变化,并分析其与炎症指标、免疫细胞亚群及细菌负荷的相关性。方法 本研究纳入90名确诊的PTB患者和45名健康对照者,进行了全面的临床评估和样本收集。采用高效液相色谱-串联质谱法(HPLC-MS/MS)测定1,25-(OH)_(2)D水平,定量实时反转录聚合酶链反应(qRT-PCR)技术测定CYP27A1 mRNA表达水平。酶联免疫吸附试验(ELISA)用于测定血清中的炎症指标,包括C-反应蛋白(CRP)、红细胞沉降率(ESR)和白细胞介素-6(IL-6)。流式细胞仪用于分析免疫细胞亚群,包括CD4+T细胞、CD8+T细胞、自然杀伤(NK)细胞和调节性T细胞(Treg)。细菌负荷通过痰液中的结核分枝杆菌(Mtb)计数来评估。结果 研究发现,与健康对照组相比,PTB患者组的1,25-(OH)_(2)D水平显著降低(P<0.01),而CYP27A1 mRNA表达水平则显著升高(P<0.01)。1,25-(OH)_(2)D水平与CRP、ESR和IL-6水平呈负相关(分别为r=-0.701,P<0.01;r=-0.465,P<0.01;r=-0.715,P<0.01),与CD4+T、CD8+T、NK、Treg细胞计数呈正相关(分别为r=0.356,P<0.01;r=0.213,P<0.05;r=0.313,P<0.05;r=0.430,P<0.01)。CYP27A1 mRNA水平与炎症指标呈正相关(分别为r=0.774,P<0.01;r=0.619,P<0.01;r=0.799,P<0.01),与CD8+T细胞计数呈负相关(r=-0.230,P<0.05)。此外,1,25-(OH)_(2)D水平与细菌负荷呈负相关(r=-0.327,P<0.01),而CYP27A1 mRNA水平与细菌负荷呈正相关(r=0.430,P<0.01)。结论 1,25-(OH)_(2)D和CYP27A1 mRNA在PTB患者中的表现与健康对照组存在显著差异,并与炎症、免疫和细菌负荷具有密切的相关性。这些生物标志物可能在肺结核的发病机制和治疗反应评估中发挥作用,为未来的研究和临床应用提供了新的视角。

Objective To investigate the levels of 1,25-dihydroxyvitamin D(1,25-(OH),D)and the expression of CYP27A1 mRNA in patients with pulmonary tuberculosis(PTB)and to analyze their correlations with inflammatory markers,immune cell subsets,and bacterial load.M.ethodsThis study included 90 diagnosed PTB patients and 45 healthy controls,all of whom underwent comprehensive clinical evaluation and sample collection.The levels of 1,25-(OH),D were determined using high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS),and the expression levels of CYP27A1 mRNA were measured by quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR).Enzyme-linked immunosorbent assay(ELISA)was used to measure serum inflammatory markers,including C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),and interleukin-6(IL-6).Flow cytometry was utilized to analyze immune cell subsets,including CD4^(+)T cells,CD8^(+)T cells,natural killer(NK)cells,and regulatory T cells(Treg).Bacterial load was assessed by counting Mycobacterium tuberculosis(Mtb)in sputum.Results The study found that compared to the healthy control group,the levels of 1,25-(OH),D in the PTB patient group were significantly reduced(P<0.01),while the expression levels of CYP27A1 mRNA were significantly increased(P<0.01).The levels of 1,25-(OH),D were negatively correlated with CRP,ESR,and IL-6 levels(r=-0.701,P<0.01;r=-0.465,P<0.01;r=-0.715,P<0.01,respectively)and positively correlated with the counts of CD4^(+)T,CD8^(+)T,NK,and Treg cells(r=0.356,P<0.01;r=0.213,P<0.05;r=0.313,P<0.05;r=0.430,P<0.01,respectively).CYP27A1 mRNA levels were positively correlated with inflammatory markers(r=0.774,P<0.01;r=0.619,P<0.01;r=0.799,P<0.01,respectively)and negatively correlated with the count of CD8^(+)T cells(r=-0.230,P<0.05).Moreover,the levels of 1,25-(OH),D were negatively correlated with bacterial load(r=-0.327,P<0.01),while CYP27A1 mRNA levels were positively correlated with bacterial load(r=0.430,P<0.01).Conclusion The study reveals significant differences in the expression of 1,25-(OH),D and CYP27A1 mRNA in PTB patients compared to healthy controls and demonstrates a close correlation with inflammation,immunity,and bacterial load.These biomarkers may play a role in the pathogenesis of pulmonary tuberculosis and in the evaluation of treatment response,offering new perspectives for future research and clinical applications.