發(fā)布時間：2018-03-02 12:02

  本文選題：黃鱔　切入點：肝臟發(fā)育　出處：《江西農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：為了更好預(yù)防和治療黃鱔在養(yǎng)殖過程中出現(xiàn)的肝病,本文運用了組織學(xué)、生理生化和熒光定量PCR技術(shù),研究了黃鱔(Monopterus albus)肝臟發(fā)育過程、四氯化碳對黃鱔肝損傷及其肝組織變化、有關(guān)生理生化指標(biāo)和花生四烯酸代謝途徑中PLA2、COX-1、COX-2、5-LOX和CYP1A基因表達(dá)的影響。結(jié)果如下:(1)黃鱔肝臟的發(fā)育特點。結(jié)果發(fā)現(xiàn)黃鱔肝臟在仔魚出膜前已開始發(fā)育,后逐步完善。出膜后1d的仔鱔,消化道右側(cè)可觀察到染色較深的肝臟細(xì)胞團(tuán),3d可見肝血竇,5d的仔魚肝部開始貯存脂質(zhì),7d的仔魚肝臟積累脂質(zhì)更明顯;肝臟緊貼卵黃囊發(fā)育,隨著黃鱔肝臟的發(fā)育,卵黃囊逐漸消失殆盡并呈現(xiàn)出密切聯(lián)系,此時肝臟基本結(jié)構(gòu)已成形;出膜3個月后,已大致發(fā)育成熟;繁殖期間懷卵雌鱔肝細(xì)胞體積較未懷卵的雌鱔大。非繁殖期間的黃鱔肝臟顯微結(jié)構(gòu)無明顯差別。(2)不同CCl_4濃度對黃鱔肝組織顯微結(jié)構(gòu)的影響:試驗黃鱔注射CCl_4引起的肝細(xì)胞急性變性呈現(xiàn)出以典型的空泡變性為特征的病理變化、細(xì)胞核偏位、核仁消失和核膜不完整等,而且隨著濃度的增加而增強(qiáng)。(3)不同CCl_4濃度對黃鱔血清、肝臟生化指標(biāo)影響:實驗組血清指標(biāo)與對照組相比,0.254g/kg體質(zhì)量和0.508g/kg體質(zhì)量CCl_4溶液注射6-48 h均能引起血清谷丙轉(zhuǎn)氨酶(GPT)、谷草轉(zhuǎn)氨酶(GOT)、過氧化氫酶(CAT)、乳酸脫氫酶(LDH)活性和丙二醛(MDA)含量升高;6-96h使超氧化物歧化酶(SOD)活性下降,并且GPT、GOT、CAT活性和MDA含量在12h達(dá)到最大值(P0.01),LDH在24h達(dá)到最大值(P0.01)。與對照組相比,0.254g/kg體質(zhì)量和0.508g/kg體質(zhì)量CCl_4溶液注射6-24 h均能引起肝臟中LDH活性和MDA含量極顯著升高(P0.01);使GPT、GOT、CAT和SOD活性下降,并在12h達(dá)到峰值(P0.01),肝臟指標(biāo)在12h升高或降低值最大(P0.01)。(4)不同CCl_4濃度對黃鱔花生四烯酸代謝途徑中PLA2、COX-1、COX-2、5-LOX和CYP1A基因表達(dá)的影響:通過基因克隆和熒光定量PCR技術(shù)研究發(fā)現(xiàn),黃鱔注射CCl_4溶液6hPLA2基因表達(dá)無明顯變化,COX-1、COX-2、5-LOX和CYP1A基因表達(dá)都被抑制(P0.01);12hPLA2、COX-1、COX-2、5-LOX和CYP1A基因表達(dá)呈低濃度誘導(dǎo)高濃度抑制狀態(tài);24hPLA2、COX-2和5-LOX基因表達(dá)繼續(xù)呈低濃度誘導(dǎo)高濃度抑制狀態(tài),但較12h表達(dá)量更接近對照組,COX1和CYP1A基因表達(dá)都受到抑制;48-144h,低濃度組PLA2基因表達(dá)在96h受到誘導(dǎo)(P0.05)外,PLA2、COX-1、5-LOX和CYP1A基因表達(dá)均受到抑制;288hPLA2、COX-1、COX-2、5-LOX和CYP1A基因表達(dá)量與對照組相比均無明顯變化。
[Abstract]:In order to prevent and treat liver diseases of Monopterus Albus, the liver development process of Monopterus albus, carbon tetrachloride (CCL 4) and liver tissue changes of Monopterus albus (Monopterus albus) were studied by means of histology, physiology, biochemistry and fluorescence quantitative PCR. The effects of the expression of PLA2OX-1COX-2OX-5-LOX and CYP1A genes on physiological and biochemical indexes and arachidonic acid metabolism pathway. The results were as follows: the developmental characteristics of the liver of Monopterus Albus were as follows. The results showed that the liver of Monopterus Albus had begun to develop before the larval membrane came out. On the right side of digestive tract, it was observed that the liver of the piglets with liver blood sinusoids for 5 days began to store lipid for 7 days, and the liver was close to the yolk sac, and the liver was close to the yolk sac. With the development of the liver of Monopterus Albus, the yolk sac gradually disappeared and showed close relation. The size of liver cells of female eel during reproduction was larger than that of unfertilized eel. There was no significant difference in the microstructure of liver of ricefield eel during non-reproduction. (2) the effect of different CCl_4 concentration on the microstructure of liver tissue of Monopterus Albus: the effect of CCl_4 injection on the microstructure of liver tissue of Monopterus Albus. The acute degeneration of hepatocytes showed pathological changes characterized by typical vacuolar degeneration. Nuclear aberration, nucleolus disappearance and incomplete nuclear membrane, etc., and increased with the increase of concentration. (3) different concentrations of CCl_4 were applied to the serum of Monopterus Albus. The effect of liver biochemical indexes: compared with the control group, the serum indexes of the experimental group were 0.254 g / kg body weight and 0.508 g / kg body mass CCl_4 solution injection for 6-48 h, which could cause the activity of serum glutamate-pyruvic transaminase, glutamic oxalacetic aminotransferase, catalase, catalase and lactate dehydrogenase. The activity of superoxide dismutase (SOD) decreased when the content of malondialdehyde (MDA) and malondialdehyde (MDA) increased for 6 to 96 h. In addition, the activity of cat and the content of MDA reached the maximum value at 12 h. Compared with the control group, 0.254 g / kg CCl_4 solution and 0.508g / kg body weight CCl_4 solution for 6-24 h could significantly increase the activity of LDH and the content of MDA in liver, and decrease the activities of cat and SOD in the liver. The effects of different concentrations of CCl_4 on the expression of PLA2COX-1COX-2OX-2OX-5-LOX and CYP1A genes in arachidonic acid metabolism pathway of Monopterus Albus were studied by gene cloning and fluorescence quantitative PCR. There was no significant change in the expression of PLA2 gene at 6 h after injection of CCl_4 solution into Monopterus Albus. Both the expression of COX-2 and 5-LOX genes were inhibited by P0.01HPLA2OX-2COX-2OX-5-LOX and the expression of COX-2 and 5-LOX genes continued to be inhibited by low concentration and high concentration, and the expression of COX-2 and 5-LOX continued to be inhibited by low concentration and high concentration. However, the expression of COX1 and CYP1A genes in the control group was inhibited more closely than that in the control group for 48 to 144 hours, and the expression of PLA2 gene in the low concentration group was induced at 96 hours (P0.05). The expression of PLA2OX-1OX-5-LOX and CYP1A genes were inhibited at 288h, and the expression of PLA2COX-1COX-25-LOX and CYP1A genes did not change significantly compared with the control group.
【學(xué)位授予單位】：江西農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S947.9

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