内源性硫化氢对血管紧张素Ⅱ受体1在大鼠肝纤维化中表达的影响

发布时间：2018-04-05 11:50

本文选题：肝纤维化　切入点：硫化氢　出处：《苏州大学》2014年硕士论文

【摘要】：目的：探讨内源性H2S对实验性肝纤维化大鼠AT1表达的影响，来推测内源性H2S在大鼠肝纤维化发病中的作用机制。方法：健康Wistar雄性大鼠56只，随机分为4组：正常对照组、模型组、PAG组、NaHS组，对除正常对照组以外各组大鼠建立以CCl4诱导的大鼠实验性肝纤维化模型，并对PAG组腹腔注射胱硫醚-γ-裂解酶(CSE)抑制剂DL-炔丙基甘氨酸（PAG），NaHS组腹腔注射H2S供体硫氢化钠（NaHS），正常对照组及模型组注射等量的生理盐水。各组大鼠于造模后3周、4周后处死，各组大鼠于3、4周分别，用放射免疫法检测大鼠肝纤维化指标：血清透明质酸酶（HA），层粘蛋白（LN），III型前胶原（PcIII），IV型胶原（cIV），全自动生化检测仪检测肝功能变化：血ALT，AST，白蛋白（ALB值），病变肝脏的组织学变化（HE和Masson染色病理切片），采用免疫印记法检测血管紧张素II受体1(AT1)的表达水平，以实时荧光定量RT-PCR测定胱硫醚-γ-裂解酶(CSE)的mRNA含量变化，采用去蛋白的方法测定血清中H2S含量。结果： 1）同一时间点组织学损伤评分在模型组、PAG组、NaHS组均显著高于正常对照组，PAG组明显高于模型组，NaHS组则明显低于模型组和PAG组（P＜0.05）。 2）血清透明质酸酶（HA），层粘蛋白（LN），III型前胶原（PcIII），IV型胶原（cIV），血ALT，AST水平在模型组、PAG组、NaHS组均显著高于正常对照组，PAG组明显高于模型组，NaHS组则低于模型组，白蛋白（ALB值）的变化趋势相反（P＜0.05）。 3）肝脏CSEmRNA表达水平在模型组、PAG组、NaHS组均显著高于正常对照组，PAG组明显高于模型组，，NaHS组则低于模型组（P＜0.05）。 4）肝脏AT1表达水平在模型组、PAG组、NaHS组均高于正常对照组，PAG组明显高于模型组，NaHS组则低于模型组（P＜0.05）。 5）血浆H2S含量在模型组、NaHS组均明显高于正常对照组，PAG组低于模型组，而NaHS组明显高于PAG组和模型组（P＜0.05）。结论：内源性H2S与大鼠四氯化碳实验性肝纤维化有明显相关性，在一定范围内有抗纤维化的作用，且降低了大鼠肝脏AT1的表达，推测可能通过降低AT1的表达的机制来抑制肝纤维化的发生和发展。
[Abstract]:Objective:To explore the effect of endogenous H2S on the expression of AT1 in rats with experimental hepatic fibrosis, and to speculate the mechanism of endogenous H2S in the pathogenesis of hepatic fibrosis in rats.Methods:56 healthy male Wistar rats were randomly divided into 4 groups: normal control group, model group and nahs group. Experimental hepatic fibrosis model induced by CCl4 was established in each group of rats except the normal control group.The PAG group was treated by intraperitoneal injection of cystathionine-gamma lyase (DL-propargylglycine) inhibitor DL-propargyl glycine (PAGG) NaHS group and the normal control group and the model group were injected with the same amount of normal saline.The rats in each group were killed at 3 weeks and 4 weeks after the establishment of the model, and the rats in each group were killed at 3 weeks and 4 weeks, respectively.The indexes of hepatic fibrosis in rats were detected by radioimmunoassay: serum hyaluronidase, laminin (laminin), procollagen type III, procollagen type IV, and liver function were detected by automatic biochemical instrument: serum ALTAST, Albumin's ALB value, pathological liver tissue.The expression of angiotensin II receptor 1 (AT1) was detected by immunological imprinting method.The changes of mRNA content in cystathionine-gamma lyase (CSE) were determined by real-time fluorescence quantitative RT-PCR, and the serum H2S content was determined by deproteinization method.Results:1) at the same time, the histological injury scores in the model group were significantly higher than those in the normal control group (P < 0.05), and were significantly higher than those in the model group and the PAG group (P < 0.05).3) the expression of CSEmRNA in the model group was significantly higher than that in the normal control group (P < 0.05).4) the expression of AT1 in the model group was significantly higher than that in the normal control group (P < 0.05).5) the plasma H2S content in the model group was significantly higher than that in the normal control group (P < 0.05), while that in the NaHS group was significantly higher than that in the PAG group and the model group (P < 0.05).Conclusion:Endogenous H2S has a significant correlation with experimental hepatic fibrosis of carbon tetrachloride in rats, and has anti-fibrosis effect to a certain extent, and decreases the expression of AT1 in rat liver.It is speculated that the pathogenesis and development of hepatic fibrosis may be inhibited by decreasing the expression of AT1.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.2

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