AIH相关因子对HepG2细胞ASGPR合成的影响

发布时间：2018-04-09 08:41

本文选题：去唾液酸蛋白受体　切入点：自身免疫性肝炎　出处：《郑州大学》2015年硕士论文

【摘要】：背景:自身免疫性肝炎(autoimmune hepatitis,AIH)是与机体自身免疫相关的,病因和发病机制尚未研究清楚的肝脏疾病。研究认为,自身免疫性肝炎肝损伤主要机制涉及两个方面:T细胞介导的细胞毒性作用和抗体依赖性细胞介导的细胞毒性作用。但以哪种研究机制为主,尚未定论。而细胞因子的变化贯穿整个损伤机制。近年来,大量研究发现,肝细胞表面的去唾液酸蛋白受体(Asialoglycoprotein receptor,ASGPR)与自身免疫性肝炎发病有很大关联。Hep G2细胞的去唾液酸蛋白受体在IL-1、IL-6等不同细胞因子的刺激下合成的数量和配体结合活性会发生变化。目前,关于自身免疫性肝炎相关白介素IL-2、IL-12、TNF-α对去唾液酸蛋白受体合成的影响报道较少。去唾液酸蛋白受体在自身免疫性肝炎相关白介素的影响下,对其合成的影响分析,无疑能提高去唾液酸蛋白受体对自身免疫性肝炎肝损伤机制的认识。目的:研究自身免疫性肝炎相关白介素IL-2、TNF-α、IL-12对Hep G2细胞合成ASGPR的影响,从而为自身免疫性肝炎的肝损伤机制提供新线索。方法:1.采用RT-PCR方法检测Hep G2细胞在TNF-a、IL-2和IL-12刺激2h、20h、24h后的ASGPR1m RNA和ASGPR2 m RNA表达量变化。2.Western Blot检测Hep G2细胞在TNF-α、IL-2、IL-12刺激后2h、20h、24h后合成表面ASGPR蛋白变化。3.免疫组织化学法检测AIH患者肝细胞表面ASGPR表达情况。结果:1.TNF-α可同步作用于ASGPR1m RNA和ASGPR2 m RNA,2h与20h表达量不同,差异有统计学意义(P1=0.002,P2=0.000);IL-2不能同步作用ASGPR1m RNA和ASGPR2 m RNA,ASGPR1m RNA呈先下降后上升趋势,20h与24h表达量差异有统计学意义(P=0.046),ASGPR2 m RNA呈先上升后下降趋势,2h与20h、2h与24h表达量差异均有统计学意义(P=0.001、P=0.000);IL-12不能同步作用ASGPR1m RNA和ASGPR2 m RNA,ASGPR1m RNA呈先下降趋势,2h与24h表达量差异有统计学意义(P=0.042),ASGPR2 m RNA呈先上升后下降趋势,表达量无差异(χ2=3.289,P=0.193)。2.Hep G2细胞表面ASGPR蛋白表达量:在TNF-α刺激下。24h内呈现上升趋势(P0.05);在IL-2刺激下,24h内呈现下降趋势(P0.05);在IL-12刺激下,呈先上升后下降趋势(P0.05)。3.AIH患者肝细胞ASGPR表达减少,胆小管表面出现表达。结论:1.细胞因子TNF-α、IL-2、IL-12可直接影响ASGPR合成。肝细胞表面ASGPRS数量的变化可能参与到AIH肝损伤机制。2.TNF-α在24h内基本可同步作用于ASGPR1m RNA和ASGPR2 m RNA,IL-12、IL-2则不能同步作用于ASGPR1m RNA和ASGPR2 m RNA。
[Abstract]:Background: autoimmune hepatitis autoimmune hepatitis (AIH) is a liver disease associated with autoimmune, etiology and pathogenesis.It is suggested that the mechanism of liver injury in autoimmune hepatitis involves two aspects: the cytotoxicity mediated by T cells and the cytotoxicity mediated by antibody dependent cells.However, which kind of research mechanism is the main, has not yet been concluded.The change of cytokines throughout the damage mechanism.In recent years, a large number of studies have found thatThe sialic acid protein receptor Asialoglycoprotein receptor (ASGPRR) on the surface of hepatocytes is closely related to the pathogenesis of autoimmune hepatitis. The amount and ligand binding activity of sialic acid protein receptor in Hep G2 cells stimulated by different cytokines such as IL 1 and IL 6 may change.At present, there are few reports about the effect of IL-12 TNF- 伪 on sialic acid receptor synthesis in autoimmune hepatitis.The effect of sialic acid protein receptor on the synthesis of sialic acid protein receptor under the influence of autoimmune hepatitis related interleukin can undoubtedly improve the understanding of the mechanism of liver injury caused by sialic acid protein receptor in autoimmune hepatitis.Aim: to study the effect of interleukin-2 (IL-2TNF- 伪) IL-12 on the synthesis of ASGPR in Hep G2 cells, and to provide a new clue for the mechanism of liver injury in autoimmune hepatitis.Method 1: 1.The expression of ASGPR on hepatocytes in patients with AIH was detected by immunohistochemical method.Results 1. TNF- 伪 could simultaneously act on ASGPR1m RNA and ASGPR2 m RNAs for 2 h and 20 h respectively.There was expression on the surface of the bile duct.Conclusion 1.Cytokine TNF- 伪 and IL-2 IL-12 can directly affect the synthesis of ASGPR.The changes of ASGPRS number on the surface of hepatocytes may be involved in the mechanism of AIH liver injury. 2. TNF- 伪 can act synchronously on ASGPR1m RNA and ASGPR2 m RNA-IL-12 / IL-2 within 24 hours, but not on ASGPR1m RNA and ASGPR2 m RNA.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R575.1

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