四氯化碳对黄鳝肝损伤及其花生四烯酸代谢相关基因表达量的影响

发布时间：2018-03-02 12:02

本文选题：黄鳝　切入点：肝脏发育　出处：《江西农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：为了更好预防和治疗黄鳝在养殖过程中出现的肝病,本文运用了组织学、生理生化和荧光定量PCR技术,研究了黄鳝(Monopterus albus)肝脏发育过程、四氯化碳对黄鳝肝损伤及其肝组织变化、有关生理生化指标和花生四烯酸代谢途径中PLA2、COX-1、COX-2、5-LOX和CYP1A基因表达的影响。结果如下:(1)黄鳝肝脏的发育特点。结果发现黄鳝肝脏在仔鱼出膜前已开始发育,后逐步完善。出膜后1d的仔鳝,消化道右侧可观察到染色较深的肝脏细胞团,3d可见肝血窦,5d的仔鱼肝部开始贮存脂质,7d的仔鱼肝脏积累脂质更明显;肝脏紧贴卵黄囊发育,随着黄鳝肝脏的发育,卵黄囊逐渐消失殆尽并呈现出密切联系,此时肝脏基本结构已成形;出膜3个月后,已大致发育成熟;繁殖期间怀卵雌鳝肝细胞体积较未怀卵的雌鳝大。非繁殖期间的黄鳝肝脏显微结构无明显差别。(2)不同CCl_4浓度对黄鳝肝组织显微结构的影响:试验黄鳝注射CCl_4引起的肝细胞急性变性呈现出以典型的空泡变性为特征的病理变化、细胞核偏位、核仁消失和核膜不完整等,而且随着浓度的增加而增强。(3)不同CCl_4浓度对黄鳝血清、肝脏生化指标影响:实验组血清指标与对照组相比,0.254g/kg体质量和0.508g/kg体质量CCl_4溶液注射6-48 h均能引起血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)、过氧化氢酶(CAT)、乳酸脱氢酶(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达到最大值(P0.01),LDH在24h达到最大值(P0.01)。与对照组相比,0.254g/kg体质量和0.508g/kg体质量CCl_4溶液注射6-24 h均能引起肝脏中LDH活性和MDA含量极显著升高(P0.01);使GPT、GOT、CAT和SOD活性下降,并在12h达到峰值(P0.01),肝脏指标在12h升高或降低值最大(P0.01)。(4)不同CCl_4浓度对黄鳝花生四烯酸代谢途径中PLA2、COX-1、COX-2、5-LOX和CYP1A基因表达的影响:通过基因克隆和荧光定量PCR技术研究发现,黄鳝注射CCl_4溶液6hPLA2基因表达无明显变化,COX-1、COX-2、5-LOX和CYP1A基因表达都被抑制(P0.01);12hPLA2、COX-1、COX-2、5-LOX和CYP1A基因表达呈低浓度诱导高浓度抑制状态;24hPLA2、COX-2和5-LOX基因表达继续呈低浓度诱导高浓度抑制状态,但较12h表达量更接近对照组,COX1和CYP1A基因表达都受到抑制;48-144h,低浓度组PLA2基因表达在96h受到诱导(P0.05)外,PLA2、COX-1、5-LOX和CYP1A基因表达均受到抑制;288hPLA2、COX-1、COX-2、5-LOX和CYP1A基因表达量与对照组相比均无明显变化。
[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.
【学位授予单位】：江西农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S947.9

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