ADAMTS-1在大鼠肺纤维化组织中的表达及苦参素的干预作用

发布时间：2019-03-17 14:18

【摘要】：目的：探讨含I型血小板结合蛋白基序的解聚素样基质金属蛋白酶（ADAMTS-1）在大鼠肺纤维化组织中表达的动态变化以及苦参素的干预效果，阐明ADAMTS-1参与肺纤维化的分子机制及苦参素抗肺纤维化的机制。方法：60只SD大鼠随机分成4组，即对照组、模型组、低剂量(50mg/kg)苦参素干预组，高剂量(100mg/kg)苦参素干预组。每组15只，各组分别于第7、14、和28天处死5只大鼠，采用HE染色、Masson三联染色检测肺纤维化的程度，用免疫组化和RT-PCR分别测定肺组织ADAMTS-1、I型胶原（COLI）的蛋白及mRNA水平。结果： 1、HE染色、Masson三联染色 HE染色：对照组大鼠肺泡结构正常、清晰，未见肺泡间隔增宽及异常的胶原沉积。肺纤维化模型组大鼠第7天表现为明显的急性炎症期，可见大量炎性细胞浸润；14天时肺间隔明显增宽，胶原沉积增多及斑片状的纤维化改变；28天可见大量增生斑片状胶原纤维，肺组织结构紊乱，少许炎性细胞浸润，稳定的肺纤维化形成。 Masson染色：胶原纤维呈现蓝绿色，14天及28天可见大量的胶原纤维聚集及沉积。对照组大鼠支气管壁及肺间质局部可见散在的胶原纤维。而高剂量苦参素干预组比低剂量苦参素干预组及模型组胶原纤维的聚集沉积明显有所减轻，低剂量苦参素干预组与模型组相比胶原纤维的分布及量略少。 2. HYP含量检测随着肺纤维化模型建造时间的递增HYP含量越高，模型组HYP含量与对照组相比差异有显著性（P0.05），与低剂量苦参素组相比差异无显著性（P＞0.05），与高剂量苦参素组相比差异有显著性（P 0.05）。 3.ADAMTS-1蛋白在肺纤维化中的表达 ADAMTS-1在模型组各时间点均表达，其表达量随着时间的递增呈逐渐下降趋势；在低剂量苦参素组干预组及高剂量苦参素干预组第7天、14天、28天均表达，随着时间的递增，剂量增高，其表达逐渐升高的趋势。模型组与对照组相比ADAMTS-1表达呈显著下降，差异有显著性（P0.05）;模型组与低剂量苦参素干预组相比，虽然后者ADAMTS-1虽有增高，但无显著差异性（P0.05）；模型组与高剂量苦参素干预组比，后者ADAMTS-1表达显著增高，差异有显著性（P 0.05）;高剂量苦参素与低剂量苦参素组对比，差异有显著性（P 0.05）;低剂量苦参素组于对照组相比，差异有显著性（P 0.05）。 4.肺纤维化组织RT-PCR检测ADAMTS-1及COLI mRNA的表达 ADAMTS-1mRNA在对照组表达最高，，模型组各时间点ADAMTS-1mRNA表达水平显著低于对照组（P0.05）,高剂量苦参素干预组ADAMTS-1mRNA表达水平较模型组增加（P 0.05），但仍低于对照组（P 0.05），然而低剂量苦参素组与模型组比较差异无显著性（P0.05）。COLI mRNA在模型组7天，14天、28天时均有表达，各组及各时间点间表达均差异有显著性（P0.05）；同时与对照组相比差异有显著性（P0.05），模型组28天时表达最高。结论： 1. ADAMTS-1在肺纤维化大鼠肺组织中表达降低。 2. COLI在肺纤维化大鼠肺组织中表达升高。 3.苦参素可能通过上调ADAMTS-1表达，降低COLI含量而发挥抗肺纤维化作用。
[Abstract]:Objective: To study the dynamic changes of the expression of the depolymerized substance-like matrix metalloprotease (ADAMTS-1) containing the type I platelet-binding protein motif in the rat pulmonary fibrosis and the effect of the intervention, and to clarify the molecular mechanism of ADAMTS-1 in the pulmonary fibrosis and the mechanism of the anti-pulmonary fibrosis. Methods:60 SD rats were randomly divided into 4 groups: control group, model group, low dose (50 mg/ kg) and high dose (100 mg/ kg). The lung tissue ADAMTS-1, type I collagen (COLI) and mRNA were measured by immunohistochemistry and RT-PCR. Flat. Results:1, HE staining, Mass On-on triple-staining HE staining: the alveolar structure of the control group was normal and clear, and the alveolar space was not found. Abnormal collagen deposition. The rats in the model group of the pulmonary fibrosis showed a clear acute inflammation phase on the 7th day, with a large number of inflammatory cells infiltrates; at 14 days, the lung interval was significantly increased, the collagen deposition increased and the fibrosis of the plaque was changed; a large amount of the sheet-like collagen fiber was visible on the 28-day period, The structural disorder of the lung tissue, the infiltration and the stabilization of a few inflammatory cells The formation of pulmonary fibrosis. Masson staining: The collagen fibers present a large amount of blue-green,14-day, and 28-day The aggregation and deposition of collagen fibers in the control group, the bronchial wall and the interstitial part of the lung in the control group Compared with the model group, there was a significant reduction in the aggregation and deposition of the collagen fibers in the low-dose and low-dose elements of the intervention group and the model group, compared with the model group. The distribution and quantity of the fiber are slightly less .2. The higher the content of HYP in the model group, the higher the content of HYP in the model group, the higher the content of HYP in the model group than that of the control group (P0.05). No significant difference (P> 0.05), compared with the high-dose element group. The difference was significant (P 0.05).3. ADA The expression of MTS-1 protein in the pulmonary fibrosis of ADAMTS-1 was expressed in all the time points of the model group, and the expression of MTS-1 protein gradually decreased with the increasing of time. 4-day,28-day expression, over time The expression of ADAMTS-1 in the model group was significantly lower than that of the control group (P0.05). The expression of ADAMTS-1 in the latter group was significantly higher and the difference was significant (P 0.05). Compared with the group, the difference was significant (P 0.05).4. RT-PCR detection of pulmonary fibrosis The expression of ADAMTS-1 and COLI mRNA was the highest in the control group, and the expression level of ADAMTS-1 mRNA in the model group was significantly lower than that in the control group (P0.05). Compared with the control group (P 0.05), there was no significant difference between the low dose and the model group (P 0.05). The expression of COLI mRNA in the model group at 7,14 and 28 days was significant (P0.05). (a) (a) (a The highest expression was found at 28 days in model group (P0.05). 1. ADAMTS-1 in the lung tissue of rats with pulmonary fibrosis Up to down.2. The expression of COLI in lung tissue of rats with pulmonary fibrosis was increased.
【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】：R285.5;R563.9

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