发展“快”“慢”COPD患者脏腑虚损状况与Th17/Treg细胞比例及NO合成酶SNPs相关性研究

There are two penotypes after the onset of COPD,The devolepment speed is 'Fast' i.e. rapid progress in a short condition. The other is a progression of 'Slow',that is relatively stable in the pathogenesis of the disease for a long time . The virtual organs dificiency is the internal factors of COPD,virtual Lunge,spleen and kidney participate in breathing regulator and liquid regulation .Development of the disease process and the different stages of disease severity wasting organs exhibited different circumstances, the present srudy is to investigate the correlation between the development of COPD'Fast','Slow' two phenotypes and virtual lung slpeen and kidney dificiency. NO molecules influence the differentiation of naive T cells through nuclear factor-кB signaling. Th17, Treg cells and inducible NO-mediated mediated COPD airway inflammation, affecting the pace of development of COPD disease. We using flow liquid multiple quantitative techniques, such as PCR - RELP technology to participate in the airway inflammation response of Th17 and Treg cells and participate in immune regulation NO, on patients with COPD development 'fast‘ ’slow‘ different organs is deficient status of Th17 / Treg cells and NO synthetase gene to the inner link between single nucleotide polymorphisms, looking for COPD development 'quick' or 'slow' the cause of the differences provide certain basis.

COPD患者发病后可表现出两种极端表型，一种是发展速度“快”，即在发病短时间内病情迅速进展，另一种是病情发展“慢”，即在发病很长的时间内病情相对稳定。脏腑亏虚是COPD疾病发生发展的内在原因，肺脾肾三脏参予呼吸调节和水液代谢，疾病发展过程中不同的阶段和病情轻重程度表现出脏腑虚损情况不同，本研究欲探讨COPD发展“快”“慢”两种表型与肺脾肾三脏虚损程度是否存在相关性。NO分子通过核因子-кB信号通路影响初始T细胞分化，Th17、Treg细胞及诱导性NO共同介导COPD气道的炎症反应，影响着COPD疾病的发展速度。本实验采用流式液相多重定量(BD CBA)技术、PCR-RELP技术观察Th17、Treg细胞分泌的细胞因子及诱导性NO合成酶的遗传基因，探讨COPD发展“快”“慢”患者不同脏腑虚损状况与Th17/Treg细胞比例及NO合成酶基因的单核苷酸多态性三者之间的内在联系。

COPD患者发病后可表现出两种极端表型，一种是发展速度“快”，即在短时间内病情迅速进展，另一种是病情发展“慢”，即在发病很长的时间内病情相对稳定。肺脾肾三脏参予呼吸调节和水液代谢，脏腑亏虚是COPD疾病发生发展的内在原因，疾病发展过程中不同的阶段和病情轻重程度表现出的脏腑虚损情况不同。本研究通过量表法调查发展“快”“慢”COPD患者的脏腑虚损状况，用CBA流式液相多重蛋白定量技术检测血清中IFN-γ、TNF-α、IL-17、IL-10、IL-6、IL-4、IL-2，用流式细胞术检测外周静脉血中Th17、Treg细胞，计算Th17和Treg细胞的比值，采用Greiss法检测血清中总NO含量，采用蛋白免疫印迹（Western Blot）法检测外周静脉血有核细胞中神经型一氧化氮合酶（nNOS）、诱导型一氧化氮合酶（iNOS）、内皮型一氧化氮合酶（eNOS）的含量，用ELISA法检测血清中三种NOS的含量；采用SNaPshot SNP分型技术对三种一氧化氮合成酶的9个SNP位点进行分型检测。通过研究发现发展“慢”COPD患者以肺脾虚损为主，发展“快”COPD患者以肺脾肾三藏虚损为主，且其外周血中NO含量明显增加，血液有核细胞中的iNOS含量增高，且得到结论COPD患者的iNOS基因rs3729508野生基因型CC和eNOS基因rs7830位点突变基因型（GG、GT）增加COPD病情发展“快”的风险。提示iNOS基因可能是影响COPD病情发展的靶点。