本文選題：豬繁殖與呼吸綜合征　切入點：肺損傷　出處：《華中農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：豬繁殖與呼吸綜合征(PRRS)是一種傳染性極強的豬病,臨床主要表現(xiàn)為流產(chǎn)、早產(chǎn)等繁殖障礙和不同程度的呼吸癥狀,如呼吸困難、肺炎等。病死豬剖檢通�？梢姺尾砍霈F(xiàn)彌漫性間質(zhì)性肺炎并伴有大量出血點,因此肺部病變導(dǎo)致的肺損傷是造成豬死亡的主要因素之PRRSV感染豬的過程中,肺損傷的形成可能與血管內(nèi)皮細胞形態(tài)和功能的改變關(guān)系密切。一氧化氮(NO)是可以改變內(nèi)皮細胞的形態(tài)和功能的因素之一,而內(nèi)皮型一氧化氮合酶(eNOS)控制著血管內(nèi)皮細胞中NO的合成。因此,eNOS及其相關(guān)因子的含量變化可能會與PRRSV感染豬造成肺損傷有所關(guān)聯(lián)。本實驗旨在通過對PRRS中的eNOS及其相關(guān)因子變化的研究,闡明豬PRRS中肺臟炎癥反應(yīng)及肺損傷的相關(guān)分子機理,為PRRS的預(yù)防與治療提供思路。本實驗所用樣品均為大白豬豬肺,分為三組：正常對照豬肺樣本、臨床PRRS豬肺樣與人工攻毒PRRS豬肺樣。通過熒光定量PCR對比了eNOS與iNOS在PRRS病豬中的變化；Western blot對比了eNOS與微囊蛋白-1的含量變化；分別通過硝酸還原酶法與化學(xué)熒光法檢測了肺組織內(nèi)NO和活性氧簇(ROS)含量變化。通過蘇木精-伊紅染色和免疫組化對比血管內(nèi)皮細胞形態(tài)變化和eNOS蛋白含量變化。結(jié)果顯示,與正常對照組相比,臨床組與攻毒組中eNOS的mRNA相對表達量顯著升高(P0.05); iNOS的mRNA變化不顯著(P0.05)；eNOS蛋白表達量顯著升高,Cav-1蛋白表達量臨床組中升高；NO含量顯著降低(P0.05); ROS含量變化不顯著(P0.05)；病理組織學(xué)結(jié)果表明,與正常豬相比,PRRS病豬肺中的大血管結(jié)構(gòu)破損,毛細血管管徑變小并且數(shù)目減少；免疫組化中eNOS蛋白顯著升高(P0.05)。研究結(jié)論：1.臨床感染的PRRS病豬與攻毒的PRRS病豬在實驗結(jié)果上呈現(xiàn)出一定的符合性；2. PRRSV的感染可以導(dǎo)致病豬肺內(nèi)的NO含量下降,而eNOS的mRNA和蛋白均上升,表明PRRSV感染可能導(dǎo)致eNOS的活性降低,臨床PRRS中與eNOS結(jié)合的Cav-1蛋白增加,可能是eNOS的活性降低的因素之一。3.iNOS和ROS在PRRS病豬肺組織中的變化個顯著,說明在PRRS中,iNOS對NO的調(diào)節(jié)作用并不明顯,NO減少的同時并未造成ROS堆積。
[Abstract]:Porcine reproductive and respiratory syndrome (PRRSs) is a highly infectious disease of pigs. Its clinical manifestations are abortion, premature birth and other reproductive disorders, as well as various respiratory symptoms, such as dyspnea. Lung injury caused by lung disease is the main factor leading to pig death. PRRSV infection is the main cause of pig death. The formation of lung injury may be closely related to the changes in the morphology and function of vascular endothelial cells. Nitric oxide (no) is one of the factors that can change the morphology and function of endothelial cells. Endothelial nitric oxide synthase (Enos) controls the synthesis of no in vascular endothelial cells. Therefore, the changes of Enos and its related factors may be associated with lung injury in pigs infected with PRRSV. This study was designed to investigate the effect of eNOS in PRRS on lung injury in pigs. And its related factors, To elucidate the molecular mechanism of pulmonary inflammation and lung injury in pig PRRS, and to provide ideas for the prevention and treatment of PRRS. The samples used in this study were all large white pig lungs, which were divided into three groups: normal control pig lung samples. The changes of eNOS and iNOS in PRRS infected pigs were compared by fluorescence quantitative PCR. The contents of eNOS and microcapsule protein 1 were compared by Western blot. The contents of no and Ros in lung tissue were detected by nitrate reductase method and chemical fluorescence method respectively. The morphological changes and eNOS protein contents of vascular endothelial cells were compared by hematoxylin eosin staining and immunohistochemistry. Compared with the normal control group, The relative expression of eNOS in clinical group and drug attack group was significantly higher than that in control group (P 0.05), but the mRNA expression of iNOS was not significant. The expression of Cav-1 protein was significantly increased in clinical group. The content of no was significantly decreased in clinical group, the content of ROS was not changed significantly (P 0.05); the pathological changes were not significant. Histologically, Compared with the normal pigs, the pulmonary macrovascular structure of PRRS pigs was damaged, and the diameter of capillaries became smaller and the number of capillaries decreased. The expression of eNOS protein increased significantly in immunohistochemistry. Conclusion: 1. The clinical infected PRRS pigs and the infected PRRS infected pigs showed a certain conformance with the experimental results. The infection of PRRSV could lead to the decrease of no content in the lung of the infected pigs. However, both mRNA and protein of eNOS increased, indicating that PRRSV infection may lead to the decrease of eNOS activity, and the increase of Cav-1 protein binding to eNOS in clinical PRRS may be one of the factors that contribute to the decrease of eNOS activity. 3. The changes of eNOS and ROS in the lung tissue of PRRS pigs are significant. The results showed that the effect of iNOS on no in PRRS was not obvious and no was decreased and ROS accumulation was not caused.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.28

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