FIZZ1在COPD大鼠肺组织中的表达及黄芪甲苷干预的影响

发布时间：2019-02-19 16:45

【摘要】：目的观察FIZZ1和肿瘤坏死因子α（TNF-α）在慢性阻塞性肺疾病大鼠中表达水平的动态变化，探讨两者在慢性阻塞性肺疾病中的作用，同时探讨黄芪甲苷（AST）治疗COPD的可能机制。方法雄性SD大鼠108只随机分为4组：正常对照组(A组)18只，COPD模型组30只(B组，分为模型二周组、四周组、八周组各10只)，黄芪甲苷低剂量组30只(C组，分为黄芪甲苷低剂量二周组、四周组、八周组各10只)，黄芪甲苷高剂量组30只(D组，分为黄芪甲苷高剂量二周组、四周组、八周组各10只)。B,C,D组采用每天熏吸香烟和气管内2次滴加脂多糖(LPS)联合刺激方法制作大鼠COPD模型，同时C,D组在每次熏吸香烟前30min予以黄芪甲苷1mL灌胃(用1%羧甲基纤维素钠溶解，分别含黄芪甲苷3mg,9mg)，A,B两组同时用助溶剂羧甲基纤维素钠1mL灌胃。经上述处理后，分别于二周、四周、八周3个时间段处死大鼠，常规病理切片观察各组各时间点大鼠气道和肺组织的形态学变化，免疫组织化学法检测肺组织中FIZZ1、TNF-α蛋白水平，RT-PCR法检测肺组织中FIZZ1、TNF-α mRNA的表达情况。结果 1、大鼠肺组织病理变化情况正常对照组气道管壁结构完整，周围无炎症细胞浸润，肺泡结构形态未见破坏；COPD模型二周组气道壁及平滑肌增厚，气道上皮受损，部分纤毛脱落，气道周围及肺泡间隔内有大量炎症细胞浸润；COPD模型四周组与二周组相比，气道壁及平滑肌明显增厚，管腔相对变窄，部分肺泡破坏、融合，肺泡腔扩大，仍有炎性细胞浸润；COPD模型八周组与四周组相比，气道壁及平滑肌增厚更明显，管腔更窄，大部分肺泡破坏、融合，肺泡腔扩大，，肺泡间隔断裂。经黄芪甲苷干预后，与同时间段的COPD模型组相比，气道管腔狭窄程度和肺泡结构破坏程度有减轻，且黄芪甲苷高剂量组减轻越明显。 2、大鼠肺组织FIZZ1、TNF-α mRNA和蛋白的表达水平 FIZZ1、TNF-α mRNA和蛋白在正常肺组织中表达较弱，COPD模型组FIZZ1、TNF-αmRNA和蛋白随着刺激时间的延长，表达呈增加趋势，均高于对照组（P0.05）；COPD模型二周组支气管肺组织中FIZZ1、TNF-α mRNA和蛋白表达水平增加，四周组FIZZ1、TNF-α mRNA和蛋白表达水平升高较明显，两者相比，差异有显著性（P0.05）；八周组其表达水平进一步升高，与四周组相比，差异有显著性（P0.05）；经黄芪甲苷干预后，与同一时间段的COPD模型二周、四周、八周组比较，FIZZ1、TNF-α mRNA和蛋白表达水平均有下降，且黄芪甲苷高剂量组较低剂量组下降较明显，差异有显著性（P0.05）；结论 1、FIZZ1、TNF-α在慢性阻塞性肺疾病大鼠模型中呈动态变化过程，可能参与了COPD的发生发展过程。 2、黄芪甲苷能减轻COPD气道炎症和重塑，延缓COPD的进程，且呈量效相关性，其机制可能与下调FIZZ1、TNF-α的表达有关。
[Abstract]:Objective to observe the dynamic changes of FIZZ1 and tumor necrosis factor 伪 (TNF- 伪) expression in chronic obstructive pulmonary disease (COPD) rats, and to explore the role of them in chronic obstructive pulmonary disease (COPD). To explore the possible mechanism of astragaloside (AST) in the treatment of COPD. Methods 108 male SD rats were randomly divided into 4 groups: normal control group (group A, n = 18), COPD model group (group B, n = 30), model group (n = 10, n = 10) and low-dose group (group C, n = 30). Astragaloside A (Astragaloside A) group (n = 10) was divided into four groups (n = 10) and Astragaloside A group (n = 30, n = 10, n = 10). In group D, the rat model of COPD was established by cigarette smoking and intratracheal administration of lipopolysaccharide (LPS) twice a day, and the rats in group C were treated with 30min (1% carboxymethyl cellulose sodium dissolved) before each cigarette smoking. Astragaloside A (3 mg / g) and group A B were given intragastric administration with sodium carboxymethylcellulose (1mL) as a cosolvent at the same time. After the above treatment, the rats were killed at 3 times of 2, 4 and 8 weeks, respectively. The morphologic changes of airway and lung tissues were observed by routine pathological sections and FIZZ1, in lung tissues were detected by immunohistochemical method. The level of TNF- 伪 protein and the expression of FIZZ1,TNF- 伪 mRNA in lung tissue were detected by RT-PCR method. Results 1. The pathological changes of lung tissue in the control group were characterized by intact airway wall structure, no infiltration of inflammatory cells and no destruction of alveolar structure. In COPD model group, the airway wall and smooth muscle were thickened, airway epithelium was damaged, some cilia were removed, and a large number of inflammatory cells were infiltrated around the airway and in the alveolar septum. Compared with the two-week group, the airway wall and smooth muscle were thicker, the lumen became narrower, some alveoli were destroyed, fused, the alveolar cavity was enlarged, and inflammatory cells were still infiltrated. Compared with the four groups, the airway wall and smooth muscle were thicker, the lumen was narrower, most of the alveoli were destroyed, fused, the alveolar cavity was enlarged and the alveolar septum was broken. Compared with the COPD model group at the same time, the degree of airway stenosis and the destruction of alveolar structure were reduced after the intervention of Astragaloside, and the more obvious was the abatement in the high dose group of Astragaloside A. 2, the expression level of FIZZ1,TNF- 伪 mRNA and protein in rat lung tissue was weak in normal lung tissue. FIZZ1,TNF- 伪 mRNA and protein expression in COPD model group increased with the prolongation of stimulation time. All of them were higher than the control group (P0.05). The expression of FIZZ1,TNF- 伪 mRNA and protein in bronchopulmonary tissue was increased in COPD model group for two weeks, and the expression level of FIZZ1,TNF- 伪 mRNA and protein in four groups was significantly higher than that in control group (P0.05). The expression level of eight-week group was further increased compared with the four-week group the difference was significant (P0.05). After the intervention of Astragaloside A, the expression of FIZZ1,TNF- 伪 mRNA and protein decreased significantly in the high dose group than in the low dose group, compared with the two, four and eight weeks groups of COPD model in the same time period, and the expression of FIZZ1,TNF- 伪 mRNA and protein in the high dose group was significantly lower than that in the low dose group. The difference was significant (P0.05). Conclusion\ 2. Astragaloside A can reduce airway inflammation and remodeling of COPD and delay the progress of COPD, which may be related to the down-regulation of FIZZ1,TNF- 伪 expression.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R563.9

【参考文献】