基于代谢组学益气通窍饮对慢性鼻-鼻窦炎黏膜重塑影响的动态研究

Chronic rhinitis-sinusitis is a nasal of chronic inflammatory disease, a higher incidence and recurrence rates.It is the most common and one of common chronic systemic diseases. Metabonomics technologies used in this study, an ultra-high performance liquid-mass spectrometry metabolomic analyzer as the core, the body's metabolic pattern recognition and the organic combination of modern analytical technology, the determination of lung qi deficiency type cold CRS patients with blank control group in the nasal mucosa, peripheral blood and specificity of endogenous metabolites in urine, and oral “Yiqitongqiao Yin ”after a nonspecific changes of the endogenous metabolites group; Used at the same time HE and special staining to evaluate local tissue mucosa CRS nasal mucosa remodeling, immune histochemical method to detect patients before treatment affect organizational reshaping of cytokines; Looking for nasal mucosa, blood, urine metabolites targets associated with mucosal remodeling, dynamic observation of related metabolic product in the change of “YiqitongqiaoYin” after the intervention. From the level of metabolomics reveals the material basis of TCM nasal disease differentiation, the correlation between specific endogenous metabolites and mucosa remodeling; Explore the therapeutic effect of“ Yiqitongqiao Yin ”of CRS mechanism, which will provide a scientific basis for the widespread use of traditional Chinese medicine treatment of CRS.

慢性鼻-鼻窦炎系常见病, 中医辨证以肺脾气虚为本居多，前期研究表明“益气通窍饮”具有较好的治疗作用，但其病理代谢变化及中药作用机制尚未明了。因此提出假说“益气通窍饮”可能通过调节代谢产物治疗CRS。本研究应用代谢组学技术，以超高液相-质谱联用代谢组分析仪为核心手段，将机体代谢模式识别与现代分析技术有机结合，测定肺脾气虚型CRS病人与胆腑郁热型及空白对照组鼻腔黏膜、外周血及尿液中特异性内源性代谢产物的差异，及口服“益气通窍饮”后特异性内源性代谢物的变化情况；同时使用HE及特殊染色对CRS鼻腔局部组织黏膜重塑进行评价，免疫组织化学法检测治疗前病人影响组织重塑的细胞因子；寻找其代谢物与黏膜重塑相关的靶点，动态观察代谢产物在“益气通窍饮”干预后的变化。从代谢组学水平进一步揭示CRS病理变化及中药作用机制的辨证物质基础、特异性内源性代谢物与黏膜重塑的相关性，为阐明中医鼻病辨证理论及治疗提供科学依据。

慢性鼻-鼻窦炎系常见病, 中医辨证以肺脾气虚为本且居多，本研究募集了32例中医辨证为肺脾气虚型的CRSsNP患者、20例胆腑郁热型RS患者，以及20例正常对照受试者。肺脾气虚型CRSsNP患者口服益气通窍饮八周后症状体征显著改变，总有效率为93.75%。与空白对照组相比，胆腑郁热组及肺脾气虚组鼻腔黏膜腺体增生，分泌旺盛，杯状细胞增生，基底膜区及黏膜内的血管腺体周围胶原沉积增加，基底膜厚度明显增厚，鼻腔黏膜存在明显的重塑改变。免疫组织化学染色结果显示胆腑郁热组及肺脾气虚组鼻腔黏膜中TGF-β1、MMP7、TIMP-1有不同程度的阳性表达。经益气通窍饮治疗后，肺脾气虚组鼻腔黏膜腺体、杯状细胞较疗前数量减少，胶原纤维实验后沉积减少，而基底膜厚度实验前后无明显变化，TGF-β1因子呈灶状弱阳性表达，MMP7蛋白呈弥漫弱阳性表达，TIMP-1呈中等阳性表达。在胆腑郁热型组与空白对照组差异代谢物中，筛选出血清、尿液、黏膜样本中具有重复性的代谢产物：Oxoglutaric acid、LysoPC(20:4(8Z,11Z,14Z,17Z))、PC(18:2(9Z,12Z)/16:0)、PC(18:0/18:2(9Z,12Z))、PE(P-18:1(11Z)/22:6(4Z,7Z,10Z,13Z,16Z,19Z))、LysoPE(18:0/0:0)含量呈升高趋势，参与D -谷氨酰胺和D -谷氨酸代谢；精氨酸生物合成；丁酸甲酯代谢；TCA循环；丙氨酸、天门冬氨酸和谷氨酸代谢，说明胆腑郁热型RS会引起上述代谢物和代谢通路的改变。在肺脾气虚型组与空白对照组差异代谢物中：L-Glutamine含量呈下降趋势，PC(18:2(9Z,12Z)/16:0)；LysoPE(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/0:0)含量呈上升趋势，参与D -谷氨酰胺和D -谷氨酸代谢；精氨酸生物合成；丙氨酸、天门冬氨酸和谷氨酸代谢；氨酰基-tRNA 合成。肺脾气虚型CRSsNP组经益气通窍饮治疗四周后代谢轮廓向空白对照组靠近，并且随着用药时间的增加，在用药八周后代谢指纹图谱变动趋势更为明显，两组间的差异性代谢产物均显示回调，表明益气通窍饮可能是通过调节上述代谢物和代谢通路以改善鼻部症状。