【摘要】：目的：探討含I型血小板結(jié)合蛋白基序的解聚素樣基質(zhì)金屬蛋白酶（ADAMTS-1）在大鼠肺纖維化組織中表達(dá)的動(dòng)態(tài)變化以及苦參素的干預(yù)效果，闡明ADAMTS-1參與肺纖維化的分子機(jī)制及苦參素抗肺纖維化的機(jī)制。 方法：60只SD大鼠隨機(jī)分成4組，即對(duì)照組、模型組、低劑量(50mg/kg)苦參素干預(yù)組，高劑量(100mg/kg)苦參素干預(yù)組。每組15只，各組分別于第7、14、和28天處死5只大鼠，采用HE染色、Masson三聯(lián)染色檢測(cè)肺纖維化的程度，用免疫組化和RT-PCR分別測(cè)定肺組織ADAMTS-1、I型膠原（COLI）的蛋白及mRNA水平。 結(jié)果： 1、HE染色、Masson三聯(lián)染色 HE染色：對(duì)照組大鼠肺泡結(jié)構(gòu)正常、清晰，未見(jiàn)肺泡間隔增寬及異常的膠原沉積。肺纖維化模型組大鼠第7天表現(xiàn)為明顯的急性炎癥期，可見(jiàn)大量炎性細(xì)胞浸潤(rùn)；14天時(shí)肺間隔明顯增寬，膠原沉積增多及斑片狀的纖維化改變；28天可見(jiàn)大量增生斑片狀膠原纖維，肺組織結(jié)構(gòu)紊亂，少許炎性細(xì)胞浸潤(rùn)，穩(wěn)定的肺纖維化形成。 Masson染色：膠原纖維呈現(xiàn)藍(lán)綠色，14天及28天可見(jiàn)大量的膠原纖維聚集及沉積。對(duì)照組大鼠支氣管壁及肺間質(zhì)局部可見(jiàn)散在的膠原纖維。而高劑量苦參素干預(yù)組比低劑量苦參素干預(yù)組及模型組膠原纖維的聚集沉積明顯有所減輕，低劑量苦參素干預(yù)組與模型組相比膠原纖維的分布及量略少。 2. HYP含量檢測(cè) 隨著肺纖維化模型建造時(shí)間的遞增HYP含量越高，模型組HYP含量與對(duì)照組相比差異有顯著性（P0.05），與低劑量苦參素組相比差異無(wú)顯著性（P＞0.05），與高劑量苦參素組相比差異有顯著性（P 0.05）。 3.ADAMTS-1蛋白在肺纖維化中的表達(dá) ADAMTS-1在模型組各時(shí)間點(diǎn)均表達(dá)，其表達(dá)量隨著時(shí)間的遞增呈逐漸下降趨勢(shì)；在低劑量苦參素組干預(yù)組及高劑量苦參素干預(yù)組第7天、14天、28天均表達(dá)，隨著時(shí)間的遞增，劑量增高，其表達(dá)逐漸升高的趨勢(shì)。模型組與對(duì)照組相比ADAMTS-1表達(dá)呈顯著下降，差異有顯著性（P0.05）;模型組與低劑量苦參素干預(yù)組相比，雖然后者ADAMTS-1雖有增高，但無(wú)顯著差異性（P0.05）；模型組與高劑量苦參素干預(yù)組比，后者ADAMTS-1表達(dá)顯著增高，差異有顯著性（P 0.05）;高劑量苦參素與低劑量苦參素組對(duì)比，差異有顯著性（P 0.05）;低劑量苦參素組于對(duì)照組相比，差異有顯著性（P 0.05）。 4.肺纖維化組織RT-PCR檢測(cè)ADAMTS-1及COLI mRNA的表達(dá) ADAMTS-1mRNA在對(duì)照組表達(dá)最高，，模型組各時(shí)間點(diǎn)ADAMTS-1mRNA表達(dá)水平顯著低于對(duì)照組（P0.05）,高劑量苦參素干預(yù)組ADAMTS-1mRNA表達(dá)水平較模型組增加（P 0.05），但仍低于對(duì)照組（P 0.05），然而低劑量苦參素組與模型組比較差異無(wú)顯著性（P0.05）。COLI mRNA在模型組7天，14天、28天時(shí)均有表達(dá)，各組及各時(shí)間點(diǎn)間表達(dá)均差異有顯著性（P0.05）；同時(shí)與對(duì)照組相比差異有顯著性（P0.05），模型組28天時(shí)表達(dá)最高。 結(jié)論： 1. ADAMTS-1在肺纖維化大鼠肺組織中表達(dá)降低。 2. COLI在肺纖維化大鼠肺組織中表達(dá)升高。 3.苦參素可能通過(guò)上調(diào)ADAMTS-1表達(dá)，降低COLI含量而發(fā)揮抗肺纖維化作用。
[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.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R285.5;R563.9

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