黄芪注射液对博来霉素诱导的大鼠肺纤维化的作用研究

发布时间：2018-11-20 11:19

【摘要】：目的：本研究以气管注入博莱霉素复制大鼠肺纤维化模型，通过对与肺纤维化相关指标，包括与氧化应激相关的超氧化物歧化酶（SOD）、丙二醛（MDA）、平滑肌肌动蛋白-α（α-SMA）、转化生长因子-β1（TGF-β1）和其下游信号通路TGF-β/Smads的检测，研究探讨黄芪注射液对大鼠肺纤维化模型的影响，为肺纤维化的临床治疗开发新的药物。方法：选用Wistar雄性大鼠30只，随机均分3组，即空白对照组（生理盐水）、模型组（博莱霉素BLM）、治疗组（黄芪注射液）。治疗组和模型组大鼠分别用气管内注入BLM的方法，，制备肺纤维化模型。造模后第21天开始，以腹腔注射给药方式，治疗组给予黄芪注射液5ml/kg，模型组及空白对照组给予生理盐水5ml/kg，每日一次，连续两周。末次给药后1h，将大鼠用10%的水合氯醛（3.0ml/kg）溶液麻醉。选用腹主动脉取血的方式采集血液样本，离心15min制备血清，分装备用，检测血清中SOD和MDA含量，观察各组大鼠氧化和抗氧化作用失衡的严重程度。取出的肺组织制备常规病理切片，用HE染色法观察大鼠肺组织正常细胞和病变细胞的一般形态，观察肺泡炎症程度，肺泡间隔以及有无炎性细胞浸润发生。通过Masson染色的方法观察肺组织中胶原纤维，结合HE染色结果共同分析，用以鉴定肺纤维化的程度。同时将肺组织标本，用免疫组化方法和蛋白免疫印迹法（Western blot），检测α-SMA、TGF-β1、Smad2/3在肺组织中的表达。结果：从病理切片可以看出，模型组大鼠肺组织中有明显的炎性细胞浸润和胶原纤维的增生，治疗组情况明显好转。免疫组化切片和蛋白免疫印迹结果都可以看出，与对照组相比，模型组α-SMA、TGF-β1表达量明显升高，TGF-β/Smads信号通路被激活；而治疗组α-SMA、TGF-β1含量已被有效控制，抑制了TGF-β/Smads信号传导。通过检测血清中SOD活性和MDA含量发现，与空白组比较模型组血清中SOD活性明显降低（P0.05），而MDA含量显著升高（P0.05）；与模型组相比治疗组中SOD含量明显升高（P0.05），MDA含量明显降低（P0.05）。结论：从α-SMA表达可以看出黄芪注射液可抑制成纤维细胞向肌成纤维细胞转化，对肺纤维化起到一定抑制作用。作用机制是黄芪注射液通过抑制TGF-β1的表达，并参与调控TGF-β/Smads信号转导通路的传导抑制肺纤维化；同时黄芪注射液通过增强SOD活力，减低MDA水平，可以提高机体抗氧化能力，从而达到抑制肺纤维化的目的。
[Abstract]:Objective: to establish a rat model of pulmonary fibrosis by trachea injection of bleomycin, and to investigate the relationship between pulmonary fibrosis and superoxide dismutase (SOD),) malondialdehyde (MDA),) related to oxidative stress. Smooth muscle actin 伪 (伪-SMA), transforming growth factor- 尾 1 (TGF- 尾 1) and its downstream signal pathway, TGF- 尾 / Smads) were detected to investigate the effects of astragalus injection on pulmonary fibrosis in rats. To develop new drugs for the clinical treatment of pulmonary fibrosis. Methods: thirty male Wistar rats were randomly divided into 3 groups: blank control group (normal saline) and model group (bleomycin BLM), treatment group). Pulmonary fibrosis models were established in the treatment group and the model group by intratracheal injection of BLM. From the 21st day after modeling, the treatment group was given Astragalus membranaceus injection 5 ml / kg by intraperitoneal injection, and the model group and blank control group were given normal saline 5 ml / kg once a day for two weeks. Rats were anesthetized with 10% chloral hydrate (3.0ml/kg) solution 1 h after the last administration. Blood samples were collected from abdominal aorta and centrifuged 15min was used to prepare serum. The contents of SOD and MDA in serum were measured and the severity of imbalance of oxidation and antioxidation was observed in each group. The normal cells and pathological cells were observed by HE staining, the degree of alveolar inflammation, the alveolar septum and the infiltration of inflammatory cells were observed. The collagen fibers in lung tissue were observed by Masson staining and the results of HE staining were analyzed to identify the degree of pulmonary fibrosis. At the same time, the expression of 伪-SMA,TGF- 尾 1 and Smad2 / 3 in lung tissue was detected by immunohistochemistry and Western blot (Western blot),. Results: the pathological section showed that there were obvious inflammatory cell infiltration and collagen fiber proliferation in the lung tissue of the model group, and the condition of the treatment group was improved obviously. Compared with the control group, the expression of 伪-SMA,TGF- 尾 1 in the model group was significantly increased and the TGF- 尾 / Smads signal pathway was activated. In the treatment group, 伪-SMA,TGF- 尾 1 content has been effectively controlled, inhibiting TGF- 尾 / Smads signal transduction. By detecting the activity of SOD and the content of MDA in serum, it was found that compared with the blank group, the activity of SOD in the serum of the model group decreased significantly (P0.05), while the content of MDA increased significantly (P0.05). Compared with the model group, the SOD content in the treatment group was significantly increased (P0.05), MDA content significantly decreased (P0.05). Conclusion: from the expression of 伪-SMA, it can be seen that astragalus injection can inhibit the transformation of fibroblasts to myofibroblasts and inhibit pulmonary fibrosis to some extent. The mechanism is that Astragalus injection inhibits the expression of TGF- 尾 1 and participates in regulating the conduction of TGF- 尾 / Smads signal transduction pathway to inhibit pulmonary fibrosis. At the same time, Astragalus injection can increase the antioxidant ability of the body by enhancing the activity of SOD and reducing the level of MDA, thus achieving the purpose of inhibiting pulmonary fibrosis.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R285.5

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