芪黄保肾颗粒对肾衰竭幼鼠肾小管上皮细胞E-cadherin表达的影响

发布时间：2018-01-07 13:20

本文关键词：芪黄保肾颗粒对肾衰竭幼鼠肾小管上皮细胞E-cadherin表达的影响　出处：《黑龙江省中医药科学院》2016年硕士论文　论文类型：学位论文

更多相关文章： 芪黄保肾颗粒 肾衰竭 UU OE-cadherin

【摘要】：目的:通过实验研究观察芪黄保肾颗粒对单侧输尿管结扎术导致肾衰竭幼鼠肾小管上皮细胞E-cadherin的表达,旨在探讨芪黄保肾颗粒在延缓儿童肾衰竭中的作用机理。方法:将75只体重80-120g,4周龄的SD雄性幼鼠随机分为5组:假手术组、模型组、芪黄保肾颗粒低、中、高剂量组(简称低、中、高剂量组),采用单侧输尿管结扎(UUO)法建立肾衰竭幼鼠模型,术前一天12h禁食,饮水自由,术后即开始给药灌胃。假手术组、模型组仅给予蒸馏水进行灌胃,低、中、高剂量组分别按人与动物等效剂量换算法,给予幼鼠芪黄保肾颗粒的用药剂量分别是2g/kg/d、4g/kg/d、6g/kg/d进行灌胃,一次1ml,每日早8:00到晚18:00两次灌胃,连续21天。在造模后的第21天选择每组15只幼鼠收集血液,尿液及肾脏组织后处死,每只幼鼠采取24h留尿及腹主动脉采血法,每次采血2-3ml,检测血肌酐(Scr)、血清尿素氮(BUN)、血清白蛋白(AIb)及24尿蛋白定量(24Pr),采血后迅速摘下梗阻侧肾脏,观察其外观,剥去外膜,生理盐水冲洗干净后,将肾脏组织浸于9%福尔马林药液中充分固化。通过模型幼鼠肾组织HE染色、免疫组化法观察肾脏病理形态学的变化以检测造模是否成功,同时以检测肾组织中E-cadherin的含量,观察芪黄保肾颗粒对E-cadherin表达的影响,用以确定芪黄保肾颗粒是否通过抑制E-cadherin的黏附过程实现延缓TEC损伤进展的。结果:1.与模型组相比较,芪黄保肾颗粒各剂量组幼鼠梗阻侧肾脏肿大、积水均较轻,幼鼠一般状态较好,肾衰症状及体征优于模型组。2.与模型组相比较,芪黄保肾颗粒各剂量组幼鼠的Scr、BUN均低于模型组(P0.05),芪黄保肾颗粒中剂量组幼鼠Scr、BUN水平最低,而中剂量组与高剂量组Scr、BUN水平相当,组间比较没有统计学意义(P0.05)。同时芪黄保肾颗粒组可以提高肾衰幼鼠AIb。3.与模型组比较,免疫组化半定量分析显示肾小管上皮细胞E-cadherin的表达均高于模型组。其中芪黄保肾颗粒中剂量组幼鼠肾小管上皮细胞E-cadherin表达最高(P0.05)。结论:1.芪黄保肾颗粒能明显改善肾衰竭幼鼠的肾脏病理损害,调整机体整体情况,缓解肾衰竭幼鼠的症状,同时延缓肾功能的进展。2.芪黄保肾颗粒能够上调肾衰幼鼠肾脏组织E-cadherin的表达,减少肾小管基底膜的破坏,使细胞外基质成分合成减少,从而抑制肾间质纤维化的发展。3.芪黄保肾颗粒各剂量组在一定程度上均能保护肾衰幼鼠的肾功能,降低肾小管的损害和拮抗肾间质纤维化的发生,其中芪黄保肾颗粒中剂量的效果最佳,为治疗儿童肾衰竭提供了临床依据。
[Abstract]:Objective: to observe the expression of E-cadherin in renal tubular epithelial cells of rats with renal failure induced by unilateral ureteral ligation. Objective: to investigate the mechanism of Qihuang Baoshen granule in delaying children renal failure. Methods: 75 male SD rats weighing 80-120g / 4 weeks were randomly divided into five groups: sham operation group and model group. Qi Huang Bao Shen granule is low, medium, high dose group (referred to as low, medium, high dose group), using UUO method of unilateral ureteral ligation to establish the model of kidney failure young rats, fasting 12 hours before operation, drinking water freedom. The model group was only given distilled water, the low, middle and high dose groups were converted according to the equivalent dose of human and animal. The dosage of Qihuangbaoshen granule was 2 g / kg / d 4 g / kg / d and 6 g / kg / d respectively. From 8:00 to 18:00, the rats were given orally twice daily for 21 days. 15 young rats in each group were selected to collect blood, urine and kidney tissue and were killed after 21 days. Blood samples were collected from 24 hours urine and abdominal aorta of each young rat. Blood creatinine (SCR) and serum urea nitrogen (bun) were detected. Serum albumin (AIb) and 24 urinary protein (24) were quickly removed from the obstructed kidney after blood collection, the appearance of the kidney was observed, the outer membrane was peeled off, and the saline was washed out. The kidney tissue was fully solidified in 9% formalin solution. The pathological changes of kidney were observed by HE staining and immunohistochemical method in order to detect the success of the model. At the same time, to detect the content of E-cadherin in renal tissue, to observe the effect of astragalus baoshen granule on E-cadherin expression. To determine whether astragalus baoshen granules can delay the progress of TEC damage by inhibiting the adhesion of E-cadherin. Results: 1. Compared with the model group. Qihuangbaoshen granule of each dose group of young rats obstruction side kidney swelling, hydronephrosis are light, the general state of young rats is better, renal failure symptoms and signs than model group. 2. Compared with model group. 2. The levels of Scr-bun in each dose group of Qihuangbaoshen granules were lower than those of model group (P0.05A), and the level of Scr-bun was the lowest in the middle dose group of Qihuangbaoshen granule. The level of Scrn bun in middle dose group was similar to that in high dose group. There was no significant difference between the two groups (P 0.05). At the same time, Astragalus baoshen granule group could increase the AIb.3. compared with the model group. The expression of E-cadherin in renal tubular epithelial cells was significantly higher than that in the model group, and the E-cadherin table of renal tubular epithelial cells in the middle dose group of Qihuangbaoshen granules was higher than that in the model group. The highest (. Conclusion 1. Astragalus baoshen granule can obviously improve renal pathological damage in young rats with renal failure. Adjust the overall situation of the body, alleviate the symptoms of young rats with renal failure, and delay the progress of renal function. 2. Qihuangbaoshen granule can up-regulate the expression of E-cadherin in kidney tissue of young rats with renal failure. Reduce the destruction of renal tubule basement membrane, reduce the synthesis of extracellular matrix components, so as to inhibit the development of renal interstitial fibrosis .3.Astragalus baoshen granule can protect the renal function of young rats to a certain extent. It can reduce the damage of renal tubules and antagonize the occurrence of renal interstitial fibrosis, of which the middle dose of Qihuangbaoshen granule is the best, which provides the clinical basis for the treatment of children with renal failure.
【学位授予单位】：黑龙江省中医药科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.5

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