雷公藤多苷对糖尿病肾病大鼠抗氧化应激作用的影响

发布时间：2018-03-01 06:10

  本文关键词： 糖尿病肾病 雷公藤多苷 氧化应激　出处：《新乡医学院》2014年硕士论文　论文类型：学位论文

【摘要】：背景糖尿病肾病(diabetic nephropathy, DN)为糖尿病最常见的慢性并发症之一,是引起发达国家终末期肾衰竭的主要原因,肾小球细胞外基质(extracelluar matrix,ECM)增生、基底膜增厚,’肾小管间质纤维化是其基本病理特征,最终可导致肾衰竭。近年来临床和研究证实,DN不仅在成人中的发病率增高,而且在儿童中的总体发病趋势也明显增加,因此儿童糖尿病肾病已成为严重威胁着儿童身心健康的代谢性疾病。但其具体机制尚不明确,近年来研究发现氧化应激(oxitative stress,OS)在DN发生、发展中发挥着重要作用。雷公藤多苷(tripterygium wilfordii polyglycoside, TWP)是从雷公藤植物中提取的总甙,其主要成分为环氧二萜内酯类化合物。研究表明TWP可通过抑制细胞和体液免疫,从多个环节抑制免疫应答过程。李军华等[[24]研究报道,TWP对肠黏膜组织具有抗氧化作用,那么TWP对肾组织是否具有抗氧化作用经查新国内尚未见报道。本研究以此为切入点,在建立DN大鼠模型的基础上,分析TWP对DN大鼠肾脏氧化应激反应的影响,研究TWP对DN大鼠肾脏的保护作用,为TWPDN提供实验数据和理论基础。 目的本研究拟通过建立DN大鼠模型,通过观察TWP对DN大鼠饮食、饮水、毛发、体重、精神状态等一般状况及血糖、肌酐清除率(creatinine clearance,Ccr)、尿素氮(blood urea nitrogen,BUN)、24h尿微量白蛋白排泄率(urinary albumin excretion rate,UAER)等生化指标的影响,分析TWP对DN大鼠肾组织丙二醛(Malondialdehyde, MDA)含量和谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)活性及血清过氧化氢酶(catalase, CAT)活性和血清超氧阴离子(superoxide oxygen anion, O2-)水平的影响,观察TWP对DN大鼠肾脏病理改变的影响,探讨TWP对DN大鼠肾脏保护作用,为TWP治疗DN提供实验数据和理论依据。 方法85只健康雄性(Sprague-Dawley,SD)大鼠,随机分为正常组(13只)和DN大鼠模型制作组(72只),模型制作组大鼠按52mg-kg-1左下腹腔内一次性注射(ip)链脲佐菌素(Streptozotocin,STZ)建立DN大鼠模型。3周后造模成功后,随机将DN大鼠(56只)分为模型对照组和TWP (4.5,9.0、18.0mg·kg-1)药物治疗组,每组14只,TWP治疗组给予灌胃(ig),每天1次,连续8周,等体积饮用水灌胃给予正常组和模型对照组。实验过程中观察大鼠的饮食、尿量、体重、毛发、精神状况等一般情况。实验结束前2d将大鼠置代谢笼,收集24h尿液,用于检测24h尿白蛋白排泄率。用药8周末,大鼠称重,10%水合氯醛麻醉大鼠,心脏取血离心,检测肌酐清除率(creatinine clearance, Ccr)、尿素氮(blood urea nitrogen,BUN),可见光方法检测血清CAT活性,比色法测定血清O2-水平。取双肾,称重,左肾制成10%匀浆,比色法测定GSH-Px活性,硫代巴比妥酸缩合法检MDA含量；右肾切成8mm3小块,甲醛固定,切片HE染色观察病理学改变。 结果 1.成功建立了DN大鼠模型。 2.TWP对DN大鼠一般状况的影响用药4周后,TWP9.0和18.0mg·kg-1组大鼠进食、饮水、尿量、精神等一般状况有所改善,用药8周后上述现象逐渐恢复正常。 3.TWP对各组大鼠体质量、肾质量、肾质量/体质量的影响与正常组相比,模型对照组大鼠肾质量、肾质量/体质量显著升高(P0.01),体质量显著降低(P0.01)；与模型对照组相比,TWP9.0和18.0mg·kg-1治疗组大鼠肾质量、肾质量/体质量显著降低(P0.01),体质量显著升高(P0.01)。 4.TWP对DN大鼠血糖、血BUN、Ccr和24hUAER变化的影响与正常组相比,模型对照组大鼠血糖、24hUAER、血BUN较其同时期显著升高(P0.01),Ccr显著降低(P0.01)；与模型对照组相比,TWP9.0和18.0mg·kg-1组大鼠24hUAER、血BUN、血糖较其同时期显著下降(P0.01),血Ccr显著升高(P0.01)。 5.TWP对DN大鼠血清CAT活性和O2-含量及肾组织GSH-Px活性和MDA含量的影响与正常组相比,模型对照组肾组织中GSH-Px和血清CAT活性显著降低(p0.01),血清O2-和肾组织MDA水平显著上升(p0.01)；与模型对照组相比,TWP9.0和18.0mg·kg-1组血清CAT活性和肾组织中GSH-Px活性明显升高(p0.01),各组间CAT活性和GSH-Px活性差异均有统计学意义(p0.05)；与模型对照组相比,TWP9.0和18.0mg·kg-1组血清O2-水平和肾组织MDA含量明显降低(p0.01)。 6.TWP对DN大鼠肾脏病理学改变的影响与正常组相比,模型组大鼠肾小球肥大、系膜细胞增生、肾小球变形；与模型对照组相比,TWP4.5、9.0、18.0mg·kg-1治疗组上述病理状况明显改善。 结论 1.TWP9.0和18.0mg·kg-1可使DN大鼠BUN、肾重/体重及UAER均显著降低,Ccr明显升高,其病理改变也明显减轻,表明TWP对DN大鼠肾脏有一定保护作用。 2.TWP对DN大鼠肾脏的保护机制之一可能与其增高血清CAT活性和肾组织GSH-Px活性,降低血清O2-水平和肾组织MDA含量,抑制氧化应激、增强机体抗氧化能力有关。
[Abstract]:Background diabetic nephropathy (diabetic nephropathy DN) is one of the most common chronic complications of diabetes, is a major cause of end-stage renal failure in developed countries, glomerular extracellular matrix (extracelluar, matrix, ECM) hyperplasia, basement membrane thickening, renal tubulointerstitial fibrosis is the basic pathologic feature, may eventually lead to kidney failure. Confirmed by clinical and research in recent years, DN not only increased the incidence in adults, and the overall incidence in children has increased significantly, so the children of diabetic nephropathy has become a serious threat to children's physical and mental health of metabolic diseases. But the mechanism is not clear, recent studies have found that oxidative stress (oxitative stress OS), in DN, plays an important role in the development of tripterygium glycosides (Tripterygium wilfordii, polyglycoside, TWP) is extracted from Tripterygium Glycosides in plants, the main Divided into two epoxy terpenoid lactone. The results show that TWP can inhibit cellular and humoral immunity, inhibit the immune response from many aspects. Li Junhua [[24] reported that TWP had antioxidant effect on the intestinal mucosa, then whether TWP has antioxidant on renal tissue for the investigation of new is not reported in China. Based on this as a starting point, based on the establishment of DN rat model, to analyze the effect of TWP on renal oxidative stress response in DN rats and protective effects of TWP on the kidney of DN rats, and provide experimental data and theoretical basis for TWPDN.
Purpose of this study is to establish the rat model of DN, the effect of TWP on DN rat diet, drinking water, hair, body weight, general condition and mental state of blood glucose, creatinine clearance rate (creatinine, clearance, Ccr), blood urea nitrogen (blood urea nitrogen, BUN 24h), urinary albumin excretion rate (urinary albumin excretion rate, UAER) of biochemical indicators, analysis of TWP on the content of renal tissue in DN rats (Malondialdehyde, MDA) content and glutathione peroxidase (glutathione peroxidase, GSH-Px) and the activity of serum catalase (catalase, CAT) and the activity of serum superoxide anion (superoxide oxygen, anion, O2-) level, the effect of TWP the renal pathological changes in DN rats, to investigate the renoprotective effects of TWP in DN rats, provide experimental data and theoretical basis for the treatment of DN TWP.
鏂规硶85鍙�鍋ュ悍闆勬�，

本文编号：1550729