一枝蒿有效部位对胃食管反流致气道高反应的作用机制研究

For the situation of GERD high prevalence in Xin Jiang, take the pathway of 5-HT-vagal nerve-afferent fibers neuropeptide secretion as a research point, to establish GER model of guinea-pig, selecting Xin Jiang ethnic medicine characteristics Artemisia rupestris L. for interventional experimental study. From bronchial reactivity, 5-HT and TRPV1 expression, neuropeptide secretion, inflammatory factor, etc. four target spot, to observe the pulmonary resistance, compliance and esophagus, trachea and lung tissue pathological changes, to measure leukocytes and classification in BALF. ELISA determinate NKA, NKB and SP content in BALF, and the content of NGF, TNF-α in the trachea, to detect the content of 5-HT, 5-HIAA in the trachea and esophagus by HPLC, to determinate TRPV1, 5HT4R, SERT of mRNA and protein expression level in esophagus and trachea through RT-PCR and Western Blot. Immunohistochemical technique and RT-PCR detect the expression of inflammation factors, such as TNF-α, IL-1β, IL-6, IL-8, PAF, ROS in the esophagus, trachea and lung tissue. To explore the mechanism of Artemisia rupestris L. active fraction for the multipoint control treatment of GER, and expect to provide new ideas and experimental basis for GER prevention and treatment.

针对新疆GERD患病率高的现状，以5-HT-迷走神经-传入纤维神经肽分泌途径作为研究切入点，建立豚鼠GER模型，选择具有新疆民族药特色的一枝蒿进行干预性实验研究。从气道反应性、5-HT和TRPV1表达、神经肽分泌、炎症因子等4个环节，观察肺阻力和顺应性及食管、气管和肺组织病理变化；测定BALF中白细胞及分类；ELISA测定BALF中NKA、NKB、SP含量，以及气管中NGF、TNF-α含量；HPLC检测食管、气管中5-HT和5-HIAA含量；RT-PCR和Western Blot测定食管、气管中TRPV1、5HT4R、SERT的mRNA和蛋白表达水平；免疫组化技术和RT-PCR检测食管、气管和肺组织中炎症因子TNF-α、IL-1β、IL-6、IL-8、PAF、ROS的表达。以期阐明一枝蒿有效部位多点调控治疗GER的作用机理，可望为防治GER提供新思路和实验依据。

针对新疆GERD患病率高的现状，以5-HT-迷走神经-传入纤维神经肽分泌途径作为研究切入点，建立GERD大鼠模型，选择新疆特色的一枝蒿进行干预性实验研究。从胃肠蠕动、气道反应性、神经肽分泌、5-HT和TRPV1表达、炎症因子等环节，采用肺功能激发试验测定气道阻力和肺顺应性，观察食管、气管和肺组织的病理变化；测定血液学白细胞及分类计数；ELISA测定大鼠肺泡灌洗液(BALF)中NKA、NKB、SP含量；检测血清中5-HT和5-HIAA含量；RT-PCR和Western Blot测定食管中TRPV1、5HT4R、SERT的mRNA和蛋白表达水平；RT-PCR检测肺脏组织中TNF-α、IL-1β、IL-6、LPCAT2等炎症因子的表达。结果显示：(1)一枝蒿提取物(AE)高剂量组能明显促进小鼠的胃排空功能；AE各剂量组均能促进小肠推进功能；(2)在乙酰甲胆碱激发后，AE各剂量组气道阻力增长百分比和气道顺应性百分比明显小于模型组(P<0.05)；(3)AE中、高剂量组可明显降低大鼠BALF中NKA、NKB含量(P<0.05)；AE高剂量组可明显增加肺泡灌洗液中SP含量(P<0.05)；(4)与模型组比较，AE高剂量组能明显升高GER大鼠血清中的5-HT含量和食管组织中5HT4R、SERT的蛋白表达 (P<0.05)，并降低食管中TRPV1 mRNA和蛋白的表达水平(P<0.05)；(5)AE中、高剂量组血液学白细胞明显降低(P<0.05)；AE高剂量组可明显降低肺脏和食管组织中TNF-α、IL-6、LPCAT2 的mRNA表达(P<0.05)。研究表明一枝蒿提取物能改善GER大鼠气道阻力和肺顺应性，可通过降低食管中TRPV1表达，抑制C纤维兴奋后末梢释放NKA、NKB神经肽，降低食管对酸的敏感性和食管超敏反应，使气道反应性降低，从而改善呼吸道症状。本研究初步阐明了一枝蒿多点调控治疗GERD的作用机理，为防治GERD提供新思路和实验依据。