胶囊渗透压泵控释IGFBPrP1对小鼠肝肺组织的影响

发布时间：2018-01-11 08:28

  本文关键词：胶囊渗透压泵控释IGFBPrP1对小鼠肝肺组织的影响　出处：《山西医科大学》2011年硕士论文　论文类型：学位论文

  更多相关文章： 纤维化 肝脏 肺脏 胰岛素样生长因子结合蛋白相关蛋白1 胶囊渗透压泵

【摘要】：研究背景： 胰岛素样生长因子结合蛋白相关蛋白1 (insulin-like growth-factor binding protein-related protein 1, IGFBPrP1)是一种可溶性细胞黏附糖蛋白,由肿瘤细胞、血管内皮细胞、平滑肌细胞和成纤维细胞等多种细胞分泌,Lopez-Bermejo等和Ma等的研究发现IGFBPrP1蛋白广泛表达于人体各种组织细胞并起着相当重要的作用,能够调节上皮细胞和纤维母细胞生长、增殖、黏附,促进血管形成,刺激内皮细胞产生前列环素,诱导细胞凋亡等多种功能。目前有关IGFBPrP1蛋白功能尚不是完全清楚。导师刘立新课题研究小组发现：用重组IGFBPrP1能使体外培养的肝星状细胞株HSC-T6活化,且在一定的剂量范围内随着IGFBPrP1剂量的增加HSC的活化程度逐渐增强；抗IGFBPrP1抗体通过抑制肝纤维化小鼠肝脏HSC的活化、减少ECM的生成及减少肝细胞的凋亡等途径发挥抗纤维化作用。IGFBPrP1可促进体外培养的肝星状细胞过表达TGF-β1及细胞外基质,增多的细胞外基质是由TGFβ1通过Smad3信号通路介导的。IGFBPrP1可能是肝纤维化发生发展过程中的种新的因子。 IGFBPrP1的生物学功能还有待进一步研究：一方面,腹腔注射动物给药IGFBPrP1半衰期短,有药物浓度高峰,代谢快,不能真正了解IGFBPrP1蛋白的功能；另一方面,IGFBPrP1有致肝纤维化的作用,对其他脏器是否也有影响及其可能机制是什么也尚未明确。为了阐明此问题,设计了本课题。 目的： 比较IGFBPrP1对小鼠肝、肺组织作用的异同,探讨IGFBPrP1对小鼠肝、肺组织的影响及其可能机制。 方法： 24只3周龄SPF级雄性C57 BL/6野生型小鼠,体重(12±2)g,由中国医学科学院实验动物研究所提供。随机分为3组：手术组(8只)用1%水合氯醛麻备小鼠,于小鼠后背部皮下行一0.5厘米左右横切口,组织钳由切口处皮下向小鼠后背部撑出一个胶囊大小的空间(空间应大到胶囊泵体可以有一定移动度,但不能让泵体轻易滑到小鼠侧翼),行手术植入注满100μg/ml IGFBPrP1溶液的胶囊渗透压泵(输药管释出口要朝内塞入皮下不可反向以致释出口刚好在伤口之下)。胶囊泵连续恒定释放IGFBPrP1溶液,泵容积90μ1,泵速0.11μ1/h,持续4周。假手术组(8只)行单纯对照手术切开缝合不植入胶囊泵,正常组(8只)不作处理。在第4周末禁食10h后,乙醚麻醉采集肝、肺组织用10%中性甲醛固定,石蜡包埋,切片。(1)分别对肝、肺组织石蜡切片做HE染色和苦味酸-天狼猩红染色观察肝、肺组织病理变化及胶原纤维形成情况；(2)免疫组织化学染色检测肝、肺组织中IGFBPrP1、III型胶原(Collagen III)、Smad3及p-Smad2/3的表达；(3) TUNEL法检测各组肝组织中肝细胞凋亡情况。 结果： (1)苦味酸-天狼猩红染色结果：手术组肝组织中胶原纤维含量较正常组和假手术组明显增多(0.46%±0.09%vs1.27.%±0.10%,0.45%±0.08%vs1.27.%±0.10%,F=597.797,P0.01),手术组肺组织中胶原纤维含量较其他两组差异不明显(0.35%±0.09%vs0.36%±0.12%,0.37%±0.12%vs 0.36%±0.12%,F=0.344,P0.05)。 (2)免疫组织化学染色结果：手术组肝组织中IGFBPrP1、III型胶原(CollagenIII)及p-Smad2/3的表达均较正常组和假手术组显著增强,Smad3表达未见增强。(IGFBPrP1: 0.26±0.04 vs 1.77±0.64,0.25±0.04 vs 1.77±0.64, F=44.064, P0.01; Collagen III:0.45±0.04 vs 2.59±0.29,0.43±0.05 vs 2.59±0.29, F=431.641,P0.01; p-Smad2/3:0.53±0.07 vs 2.44±0.32,0.51±0.08vs2.44±0.32, F=114.251, P0.01; Smad3:0.29±0.06 vs 0.33±0.04, 0.34±0.04 vs 0.33±0.04, F=1.928, P0.05)。手术组肺组织中IGFBPrP1 III型胶原(Collagen III)、Smad3及P-Smad2/3的表达较其他两组差异不明显(IGFBPrP1:0.32±0.05vs0.30±0.07, 0.29±0.06vs0.30±0.07, F=0.505, P0.05; Collagen III:0.93±0.07 vs 0.89±0.02,0.85±0.01 vs0.89±0.02, F=0.973, P0.05; Smad3:0.26±0.03 vs 0.25±0.04,0.24±0.09 vs 0.25±0.04, F=0.150, P0.05; p-Smad2/3:0.25±0.06 vs 0.26±0.05,0.28±0.06 vs 0.26±0.05, F=0.255, P0.05)。 (3) TUNEL法检测肝细胞凋亡结果：荧光显微镜下和普通光镜下手术组肝组织中凋亡肝细胞较正常组和假手术组明显增多(6.21%土2.83%vs 34.14%±11.04%,7.13%±2.59%vs34.14%士11.04%,F=114.251,P0.01；6.10%士2.23%vs 32.23%±10.60%,5.71%±2.87%vs32.23%±10.60%,F=114.251,P0.01)。 结论： (1)持续恒量的外源性IGFBPrP1可致小鼠肝纤维化,同等条件下对小鼠肺组织影响甚微而不致肺纤维化。 (2)IGFBPrP1对小鼠组织脏器的作用具有选择性。
[Abstract]:Research background:
Insulin-like growth factor binding protein related protein 1 (insulin-like growth-factor binding protein-related protein 1, IGFBPrP1) is a soluble cell adhesion glycoprotein by tumor cells, vascular endothelial cells, smooth muscle cells and fibroblasts into a variety of cell secretion, Lopez-Bermejo and Ma, and plays an important role in widespread expression the IGFBPrP1 protein in various human tissues and cells, can regulate the growth of epithelial cells and fibroblast proliferation, adhesion, angiogenesis, endothelial cells before the column ring element functions, inducing cell apoptosis. The IGFBPrP1 protein function is not entirely clear. Professor Liu Lixin research group found that recombinant IGFBPrP1 can make in vitro hepatic stellate cells activation of HSC-T6, and in a certain dose range with increasing dose of IGFBPrP1 The activation level of HSC increased gradually; the anti IGFBPrP1 antibody by inhibiting the activation of hepatic fibrosis in HSC mice, reduced the production of ECM and reduce the apoptosis of liver cells exerts anti fibrosis effect of.IGFBPrP1 can promote hepatic stellate cells in vitro expression of TGF- beta 1 and extracellular matrix, the extracellular matrix is increased by TGF beta 1 via Smad3 signal pathway mediated by.IGFBPrP1 may be in the process of the development of a new factor of hepatic fibrosis.
The biological function of IGFBPrP1 remains to be further studied: on the one hand, intraperitoneal injection of animal drug IGFBPrP1 has short half-life, peak drug concentration, fast metabolism and the function of IGFBPrP1 protein can not really understand; on the other hand, IGFBPrP1 induced hepatic fibrosis, to other organs have the effect and the possible mechanism is also not yet clear what in order to clarify this issue, the design of this topic.
Objective:
The effects of IGFBPrP1 on the liver and lung tissue of mice were compared, and the effect of IGFBPrP1 on the liver and lung tissue of mice and its possible mechanism were discussed.
Method锛，

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