發(fā)布時(shí)間：2018-03-09 14:44

  本文選題：氨氣　切入點(diǎn)：豬　出處：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：氨氣是規(guī)模化豬場(chǎng)中普遍存在的有毒有害氣體之一,較高濃度的氨氣會(huì)給仔豬和工作人員帶來(lái)健康隱患,而目前關(guān)于氨氣影響仔豬健康的研究尚不足。本研究選取7周齡14 kg的保育仔豬共12頭,隨機(jī)分為4組,用56 mg/m3左右的氨氣對(duì)3個(gè)試驗(yàn)組分別刺激4天、8天和12天,對(duì)照組則不作處理。刺激結(jié)束后采集仔豬肺,鼻黏膜和肝臟組織,提取總RNA,然后對(duì)這4個(gè)時(shí)期的樣品進(jìn)行轉(zhuǎn)錄組測(cè)序(RNA-seq),并探討氨氣刺激對(duì)3個(gè)組織轉(zhuǎn)錄譜的影響。主要結(jié)果:1.分別鑒定了3個(gè)組織氨氣刺激前后的差異表達(dá)基因(默認(rèn)和較小天數(shù)組比),肺和鼻黏膜一共6個(gè)比較(NC與3個(gè)處理組間的比較,3個(gè)處理組間的兩兩比較),肝臟只有1個(gè)比較(D12比NC)。統(tǒng)計(jì)顯示:肺的差異表達(dá)基因一共有289個(gè),鼻黏膜則有825個(gè),肝臟有589個(gè)(367個(gè)上調(diào)表達(dá),222個(gè)下調(diào)表達(dá))。2.構(gòu)建了肺和鼻黏膜差異表達(dá)基因的表達(dá)模式,并對(duì)每個(gè)模式的基因進(jìn)行了功能富集分析。肺的差異表達(dá)基因一共被分為18類,其中SCGB3A2,MMPs,BPIFA1,IL8,ARG2,COMP,PAI-1,COLs,CLCA1,ELANE等與肺纖維化發(fā)生和氣道黏液分泌相關(guān)的基因大部分處于cluster 1,cluster 2和cluster 3中。鼻黏膜的差異表達(dá)基因被分為了20類,其中cluster 1,cluster 2,cluster 3和cluster 6中CCDCs,CFAPs,TEKTs,DNAHs,LRRCs,SPAG17,COL11A2,ACAN,SNTN與黏膜功能、結(jié)構(gòu)有關(guān);cluster 4中SFTPs,BPIFBs與呼吸功能及防御反應(yīng)相關(guān);cluster 7中MASP1,LBP,C4BPA,IL1B,REG3G,S100As與急性炎癥和黏液分泌有關(guān);cluster 8和cluster 9的PAI-1,MMPs,KRTs還與上皮修復(fù)有關(guān)。3.肝臟差異表達(dá)基因顯著富集的通路(p≤0.05)與免疫,癌癥,肝病,氨基酸代謝及生理節(jié)奏有關(guān)。蛋白互作分析表明PIK3CA,TNF,NFKBIA,THBS1,JUN,HIF1A和FOS可能是重要的調(diào)控因子。4.鑒定了NC組和D12組間的差異可變剪切及差異表達(dá)轉(zhuǎn)錄本。結(jié)果顯示CD40、SAT1、ETFB、CLK1、CHTOP和TNFRSF1A在鼻黏膜或者肺NC組和D12處理組間具有顯著差異(FDR≤0.05)的可變剪接事件。同時(shí),分別在肺、鼻黏膜和肝臟上發(fā)現(xiàn)了數(shù)量不等的差異表達(dá)(FDR≤0.05)的轉(zhuǎn)錄本。功能分析表明以上基因大部分與免疫,細(xì)胞凋亡和癌癥有關(guān)。5.用22個(gè)樣品的轉(zhuǎn)錄組測(cè)序數(shù)據(jù)鑒定了903條新lincRNA,并用權(quán)重基因共表達(dá)網(wǎng)絡(luò)整合分析(Weighted gene co-expression network analysis,WGCNA)差異表達(dá)的linc基因和編碼基因(共3072個(gè))。結(jié)果顯示LIPG在肝臟D12特異性高表達(dá),TF在肝臟NC組特異性高表達(dá);而新lincRNA XLOC_004910則在鼻黏膜D12特異性高表達(dá),其臨近的9個(gè)編碼基因(上下游100 kb)則與免疫,糖/激素代謝,造血有關(guān),暗示該lincRNA可能參與了這些進(jìn)程。主要結(jié)論:氨氣刺激引起仔豬鼻黏膜、肺和肝臟組織大量基因上調(diào)表達(dá),其中刺激12天后差異表達(dá)基因數(shù)目最多。這些差異表達(dá)基因主要參與了呼吸道重塑、氧化應(yīng)激、炎癥和免疫應(yīng)答等生物學(xué)進(jìn)程。同時(shí)發(fā)現(xiàn)基因的選擇性剪切和lincRNA在仔豬應(yīng)對(duì)氨氣刺激過(guò)程中也可能發(fā)揮重要的調(diào)控作用。
[Abstract]:Ammonia is one of the most common toxic and harmful gases in large-scale pig farms. A higher concentration of ammonia will bring health risks to piglets and workers. In this study, 12 piglets aged 7 weeks and 14 kg were randomly divided into 4 groups, which were stimulated with ammonia for about 56 mg/m3 for 4 days, 8 days and 12 days, respectively. The control group was not treated. The lungs, nasal mucosa and liver tissues of piglets were collected after stimulation. Total RNAs were extracted, and then transcriptome sequencing was carried out on the samples in these four periods. The effects of ammonia stimulation on three tissue transcriptional profiles were investigated. The main results were as follows: 1. The differentially expressed genes were identified before and after ammonia stimulation in three tissues. The default and smaller days of array comparison, lung and nasal mucosa a total of 6 comparison between the comparison between NC and 3 treatment groups, three treatment groups of pairwise comparison, liver only one comparison of D12 than NCC.statistics show: lung differentially expressed genes a total of 289, 825 nasal mucosa, 589 liver, 367 upregulated expression, 222 down-regulated expression genes were constructed to express differentially expressed genes in lung and nasal mucosa. The differentially expressed genes in the lung were divided into 18 types. Most of the genes related to pulmonary fibrosis and airway mucus secretion were found in cluster 1 cluster 2 and cluster 3. The differentially expressed genes in nasal mucosa were classified into 20 categories, including cluster 1 cluster 2 and cluster 6. Structurally related to cluster 4, SFTPslBPIFBs are associated with respiratory function and defense response in cluster 7. MASP1LBPU C4BPABPAIL1BNREG3GN S100As is associated with acute inflammation and mucus secretion. The PAI-1MMPsKRTs associated with epithelial repair are also associated with epithelial repair. The pathway of liver differentially expressed gene enrichment is less than 0.05) and immunity, cancer, cancer, and cancer. Liver disease, Amino acid metabolism and physiological rhythm are related. Protein interaction analysis shows that PIK3CAA TNFKBIATHBS1THBS1JUNHIF1A and FOS may be important regulatory factors. 4. The differential variable shear and differential expression transcripts between NC group and D12 group were identified. The results showed that CD40SAT1ETFBBK1 CHTOP and TNFRSF1A were present in nasal mucus. There was a significant difference between the membrane or lung NC group and the D12 treatment group in the variable splicing events (FDR 鈮，

本文編號(hào)：1588977