【摘要】：目的：本研究以氣管注入博萊霉素復(fù)制大鼠肺纖維化模型，通過(guò)對(duì)與肺纖維化相關(guān)指標(biāo)，包括與氧化應(yīng)激相關(guān)的超氧化物歧化酶（SOD）、丙二醛（MDA）、平滑肌肌動(dòng)蛋白-α（α-SMA）、轉(zhuǎn)化生長(zhǎng)因子-β1（TGF-β1）和其下游信號(hào)通路TGF-β/Smads的檢測(cè)，研究探討黃芪注射液對(duì)大鼠肺纖維化模型的影響，為肺纖維化的臨床治療開(kāi)發(fā)新的藥物。 方法：選用Wistar雄性大鼠30只，隨機(jī)均分3組，即空白對(duì)照組（生理鹽水）、模型組（博萊霉素BLM）、治療組（黃芪注射液）。治療組和模型組大鼠分別用氣管內(nèi)注入BLM的方法，，制備肺纖維化模型。造模后第21天開(kāi)始，以腹腔注射給藥方式，治療組給予黃芪注射液5ml/kg，模型組及空白對(duì)照組給予生理鹽水5ml/kg，每日一次，連續(xù)兩周。末次給藥后1h，將大鼠用10%的水合氯醛（3.0ml/kg）溶液麻醉。選用腹主動(dòng)脈取血的方式采集血液樣本，離心15min制備血清，分裝備用，檢測(cè)血清中SOD和MDA含量，觀察各組大鼠氧化和抗氧化作用失衡的嚴(yán)重程度。取出的肺組織制備常規(guī)病理切片，用HE染色法觀察大鼠肺組織正常細(xì)胞和病變細(xì)胞的一般形態(tài)，觀察肺泡炎癥程度，肺泡間隔以及有無(wú)炎性細(xì)胞浸潤(rùn)發(fā)生。通過(guò)Masson染色的方法觀察肺組織中膠原纖維，結(jié)合HE染色結(jié)果共同分析，用以鑒定肺纖維化的程度。同時(shí)將肺組織標(biāo)本，用免疫組化方法和蛋白免疫印跡法（Western blot），檢測(cè)α-SMA、TGF-β1、Smad2/3在肺組織中的表達(dá)。 結(jié)果：從病理切片可以看出，模型組大鼠肺組織中有明顯的炎性細(xì)胞浸潤(rùn)和膠原纖維的增生，治療組情況明顯好轉(zhuǎn)。免疫組化切片和蛋白免疫印跡結(jié)果都可以看出，與對(duì)照組相比，模型組α-SMA、TGF-β1表達(dá)量明顯升高，TGF-β/Smads信號(hào)通路被激活；而治療組α-SMA、TGF-β1含量已被有效控制，抑制了TGF-β/Smads信號(hào)傳導(dǎo)。通過(guò)檢測(cè)血清中SOD活性和MDA含量發(fā)現(xiàn)，與空白組比較模型組血清中SOD活性明顯降低（P0.05），而MDA含量顯著升高（P0.05）；與模型組相比治療組中SOD含量明顯升高（P0.05），MDA含量明顯降低（P0.05）。 結(jié)論：從α-SMA表達(dá)可以看出黃芪注射液可抑制成纖維細(xì)胞向肌成纖維細(xì)胞轉(zhuǎn)化，對(duì)肺纖維化起到一定抑制作用。作用機(jī)制是黃芪注射液通過(guò)抑制TGF-β1的表達(dá)，并參與調(diào)控TGF-β/Smads信號(hào)轉(zhuǎn)導(dǎo)通路的傳導(dǎo)抑制肺纖維化；同時(shí)黃芪注射液通過(guò)增強(qiáng)SOD活力，減低MDA水平，可以提高機(jī)體抗氧化能力，從而達(dá)到抑制肺纖維化的目的。
[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 偽 (偽-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 偽-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 偽-SMA,TGF- 尾 1 in the model group was significantly increased and the TGF- 尾 / Smads signal pathway was activated. In the treatment group, 偽-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 偽-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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R285.5

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 楊永紅;夏春生;;特發(fā)性肺纖維化治療研究進(jìn)展[J];檢驗(yàn)醫(yī)學(xué)與臨床;2011年16期

2 陳珍旭;吳蓉易;張淑玉;李連琨;唐勇;;葛根素對(duì)實(shí)驗(yàn)性大鼠肺纖維化影響[J];臨床肺科雜志;2006年04期

3 王殿華;;骨髓間充質(zhì)干細(xì)胞移植治療急性肺損傷研究進(jìn)展[J];昆明醫(yī)科大學(xué)學(xué)報(bào);2012年06期

4 唐志宇;李天朗;陳嚴(yán);楊曙光;;鱉甲煎丸對(duì)肺纖維化大鼠肺組織中TGF-β_1表達(dá)的影響[J];四川中醫(yī);2011年01期

5 武慧;馮一中;顧振綸;林軍;周文軒;郭次儀;;三七總皂甙對(duì)實(shí)驗(yàn)性肺纖維化大鼠病理變化和轉(zhuǎn)化生長(zhǎng)因子-β1表達(dá)的影響[J];蘇州大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年01期

6 祖金池;崔娜;段斐;;特發(fā)性肺纖維化中的信號(hào)轉(zhuǎn)導(dǎo)通路[J];醫(yī)學(xué)研究與教育;2009年02期

7 段報(bào)喜;黃潤(rùn)月;儲(chǔ)永良;;細(xì)胞因子治療特發(fā)性肺纖維化[J];中國(guó)組織工程研究與臨床康復(fù);2007年10期

8 全燕;夏前明;李福祥;李鴻雁;張定濤;;三七總皂苷對(duì)大鼠肺纖維化及結(jié)締組織生長(zhǎng)因子表達(dá)的影響[J];西南國(guó)防醫(yī)藥;2009年01期

9 王昌明,黃慧,張珍祥,莫碧文,白晶,曾錦榮,蔣述科,任格,王績(jī)英;槲皮素對(duì)博萊霉素致鼠肺纖維化的防治作用[J];中國(guó)藥理學(xué)通報(bào);2000年01期

10 田宏印;黃芪化學(xué)研究及其有效成分[J];云南民族學(xué)院學(xué)報(bào)(自然科學(xué)版);1996年01期

相關(guān)博士學(xué)位論文 前1條

1 楊毅;藶桃抗纖方對(duì)肺泡Ⅱ型上皮細(xì)胞轉(zhuǎn)化的影響及作用機(jī)制[D];湖北中醫(yī)學(xué)院;2009年

本文編號(hào)：2344774