黄芪多糖降低2型糖尿病大鼠血糖水平及预防糖尿病相关并发症的实验研究

发布时间：2018-02-04 02:18

  本文关键词： 2型糖尿病 黄芪多糖 物质代谢 并发症　出处：《新乡医学院》2016年硕士论文　论文类型：学位论文

【摘要】：背景糖尿病是由于胰岛素分泌不足或(和)胰岛素抵抗所引起的一种以血糖持续高水平为特征的代谢综合征。持续较高水平的血糖可导致一系列器官或组织的损害,如脂肪代谢紊乱、肾脏、心脏、眼、神经功能损害等。黄芪多糖(Astragalus polysaecharides,APS),目前研究认为其具有抗衰老、增强免疫力、防止脂质过氧化以及改善肾功能等作用。本文通过观察APS对调节糖尿病大鼠血糖水平、改善血脂及肾脏功能的作用,进而分析APS在糖尿病治疗中的临床价值。目的通过腹腔注射40 mg/kg链脲佐菌素(Streptozotocin,STZ)和高脂高糖饲料喂养方式,建立2型糖尿病大鼠模型。密切观察给予不同干预的2型糖尿病大鼠血糖水平、血脂水平以及相关肝肾脏指标变化,进一步探讨分析黄芪多糖在治疗糖尿病及预防并发症中的临床价值。方法选择健康状况良好的6周龄Wistar大鼠共90只,雄性,体重为180±15g,均由新乡医学院医学实验动物中心提供。相同环境下分笼饲养,自由光照,充足饮食,各组进食量无明显差异。饲养室的温度保持在23±2℃,湿度维持在41%±15%。实验开始前,随机抽取60只分为B组和C组,将B组和C组大鼠采用高脂高糖饮食饲养6周,之后给予腹腔注射链脲佐菌素。通过腹腔注射40 mg/kg链脲佐菌素以及高脂高糖饲养方式建立2型糖尿病大鼠模型,造模过程中,T2DM组大鼠13.3%动物死亡(8/60),52只存活大鼠中4只血糖不达标,予以剔除,最终成模48只。并随机分为模型对照组(B组)和APS组(C组);余30只中随机抽取24只为正常对照组(A组)。C组大鼠给予APS 400 mg/kg/d灌胃,A、C组分别给予等量生理盐水。分别于实验第0、2、4、6周分别自各组随机抓取6只大鼠,代谢笼法收集24小时尿液;禁食12小时,晨起取尾静脉血检测空腹血糖值大于等于16.7mmol/l则表示造模成功。然后给予相同饲料喂养,于进食后0.5、1.0、1.5以及2.0小时再次分别检测血糖值。活体心脏采血5ml,制备血清,检测tc、tg、alt、ast、sod以及mda水平。另分别取肝、肾组织,制备石蜡切片或透射电镜切片,染色观察。采用spss19.0统计软件对相关实验数据进行分析,比较3组实验动物不同时间点各指标差异。组内比较采用配对t检验,组间两两比较采用独立样本t检验,α=0.05,p0.05表示差异有统计学意义。结果1.空腹血糖水平:随实验时间延长,b组大鼠glu水平呈缓慢上升趋势,c组大鼠glu水平则自第2周始呈下降趋势。组内比较:b组第4、6周时glu水平与第0周无显著差异(p0.05),而c组相同时间点glu水平则显著低于第0周(p0.05)。组间比较:第0、2周时,c组glu水平显著高于a组(p0.05),但与b组无显著差异(p0.05);而在第4、6周时,c组glu水平显著高于a组(p0.05),且低于b组(p0.05)。2.血脂水平的比较:第2、4、6周时,b、c组大鼠tc、tg水平均较a组明显上升(p0.05)。组内比较:第4、6周时,b、c大鼠tc、tg水平均显著高于第0周(p0.05)。组间比较:第2周时,c组大鼠tc、tg水平均高于a组(p0.05),但与b组无显著差异(p0.05);第4、6周时,c组大鼠tc、tg水平均高于a组(p0.05),且低于b组(p0.05)。3.肾功能指标的比较:尿微量白蛋白:试验期间a组大鼠尿微量白蛋白水平未见明显变化,b组大鼠尿微量白蛋白水平缓慢上升,而c组大鼠各时间点尿微量白蛋白水平均低于b组,且呈下降趋势。统计学分析发现,c组大鼠第2、4、6周尿微量白蛋白水平均显著高于a组(p0.05),但低于b组(p0.05)。肾组织形态学变化:实验第2周时,三组大鼠肾脏组织切片he染色观察未见明显异常。第4周时,b、c组大鼠肾脏组织切片he染色观察可见轻微肾小管损伤,局部炎症表现。第6周时,b组大鼠肾小管囊腔出现增大表现,形态改变;肾小管上皮细胞可见炎性肿大。c组大鼠肾小管上皮细胞可见轻微炎症表现,局部肾小管内可见较为少量蛋白聚集。A组大鼠未见明显异常。4.肝脏指标比较:血清学指标:第0、2周时B、C组大鼠ALT、AST、SOD以及MDA水平比较无显著差异(P0.05)。第4、6周时,C组大鼠ALT、AST以及MDA水平均高于A组,但低于B组;而SOD水平低于A组,但高于B组;差异均有统计学意义(P0.05)。肝组织形态学观察:HE染色:第2、4周时,三组大鼠肝脏组织切片HE染色观察未见明显异常。第0、2、4周时各组大鼠肝索排列规则,肝细胞结构完整,肝小叶结构显示清楚,肝窦存在,镜下未见肝细胞变性、坏死性病变。第6周时,B组大鼠肝组织局部肝细胞肿胀明显,细胞核变小,深染,胞内可见轻度空泡变性;部分肝小叶轮廓发生较为明显的改变,肝窦明显变窄,部分甚至消失不见,小叶间及汇管区可见血管扩张淤血。C组大鼠肝组织切片可见肝细胞轻度肿大,未见明显的肝细胞空泡样变性;肝小叶结构较为清晰,肝窦较前变窄,小叶间以及汇管区血管可见轻度扩张。透射电镜观察:实验第2、4周时,B组和C组大鼠肝细胞超微结构无明显变化,细胞器完整,各细胞器如内质网、线粒体等丰富,糖原轻度减少,基质轻度肿胀,脂滴明显可见。第6周时,B组大鼠肝细胞肿胀变形,线粒体减少且明显肿胀,部分线粒体出现空泡化,膜、嵴均有溶解,断裂现象;粗面内质网扩张、断裂,膜结构损坏。细胞内脂滴明显增加,糖原显著减少。C组大鼠损伤程度较B组为轻。A组大鼠肝细胞内粗面内质网排列整齐,核糖体较多且分布均匀。线粒体结构完整。结论1.黄芪多糖可有效降低2型糖尿病大鼠血糖水平,改善其胰岛素抵抗状态。2.黄芪多糖可降低TC、TG水平,改善2型糖尿病大鼠脂肪代谢紊乱状态。3.APS可增加肝细胞抗氧化酶的活性,促进氧自由基的清除,减少肝细胞损伤。4.黄芪多糖可减轻肝脏、肾脏组织损伤,改善肝、肾组织细胞形态和功能,降低尿微量白蛋白浓度,进而改善肝、肾功能。
[Abstract]:The background of hypoinsulinism or (and) insulin resistance caused by a sustained high level blood glucose metabolic syndrome characterized. Sustained high levels of blood sugar can lead to a series of organ or tissue damage, such as fat metabolism, kidney, heart, eye, nerve function damage. Astragalus Polysaccharide (Astragalus polysaecharides, APS), the present study finds that it has anti-aging, enhance immunity, prevent lipid peroxidation and improve renal function. The effect of APS on the regulation of blood glucose levels in diabetic rats, improve blood lipid and renal function, and analyze the clinical value of APS in the treatment of diabetes by intraperitoneal injection. 40 mg/kg streptozotocin (Streptozotocin, STZ) and high fat diet, type 2 diabetic rat model was established. Close observation of type 2 diabetes mellitus treated with the intervention The blood glucose level, blood lipid level and liver kidney index changes, to further explore the clinical value of Astragalus Polysaccharide in the treatment and prevention of diabetes complications. Methods 6 week old Wistar rats in good health were 90, male, weight was 180 + 15g, were provided by the Xinxiang Medical University medical experimental animal center of the same. Under the environment of the breeding, free light, adequate intake of each diet, no significant difference. The room temperature was maintained at 23 - 2 DEG C, humidity is maintained at the beginning of 41% + 15%. before the experiment, 60 rats were randomly selected and divided into B group and C group, B group and C group rats were fed with high fat high sugar diet for 6 weeks, after intraperitoneal injection of streptozotocin. Type 2 diabetic rat model was established by intraperitoneal injection of 40 mg/kg STZ and high fat and high sugar feeding method, the modeling process, the rats in group T2DM (8/60), 13.3% animal deaths 52 survival 4 rats in blood glucose is not up, to be removed, and finally die. Only 48 were randomly divided into model control group (B group) and APS group (group C); more than 30 of 24 rats were randomly selected as normal control group (group A).C rats were given intragastric administration of APS 400 mg/kg/d, A, C group were given normal saline respectively. In the experiment of 0,2,4,6 weeks respectively were random samples of 6 rats were collected 24 hours urine metabolic cage method; fasting for 12 hours, the morning from tail vein blood glucose value is greater than or equal to 16.7mmol/l saidmodeling success. Then given the same diet, in after eating 0.5,1.0,1.5 and again 2 hours respectively to detect blood glucose levels. In vivo heart blood 5ml, serum preparation, detection of TC, TG, alt, AST, SOD and MDA respectively. The other level of liver, kidney, preparation of paraffin sections and electron microscopy, staining. The experimental data were analyzed by spss19.0 software, 姣旇緝3缁勫疄楠屽姩鐗╀笉鍚屾椂闂寸偣鍚勬寚鏍囧樊寮，

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