葡萄籽原花青素对自发性高血压大鼠肾脏结构和泌尿功能的影响

发布时间：2018-06-12 11:54

本文选题：葡萄籽原花青素 + 自发性高血压大鼠　；参考：《遵义医学院》2016年硕士论文

【摘要】：目的:研究葡萄籽原花青素(GSP)对自发性高血压大鼠(SHR)肾脏结构和泌尿功能的影响并探讨其可能机制。方法:8周龄雄性SHR 24只适应性喂养1 w后随机分成4组(n=6):SHR组(SHR)、GSP低剂量治疗组(GSP-low,50 mg/kg)、GSP高剂量治疗组(GSPhigh,200 mg/kg)和卡托普利阳性对照治疗组(captopril,30 mg/kg);6只同龄雄性Wistar-Kyoto大鼠设为正常对照组(control)。按照实验分组,给大鼠灌胃上述对应剂量药物及溶媒(2 ml/次/d),每2 w测量一次各组大鼠尾动脉收缩压(SBP)。6 w后,用代谢笼收集各组大鼠24 h尿液,测定24 h尿蛋白、视黄醛结合蛋白(RBP)和α1微球蛋白(α1-MG)含量。摘眼球取血,收集血清,测定血尿素氮(BUN),血肌酐(SCr)及胱抑素C(Cys C)含量。摘取大鼠两侧肾脏,肉眼观察肾脏大体形态,称量大鼠体重及左、右肾脏质量,计算肾脏质量指数(RMI)。HE和Masson染色下观察肾脏组织形态和胶原含量变化并测定肾内小动脉中膜与血管内径比值(MT/LD)、肾小球胶原纤维沉积评分(GCDS)及肾小管间质病变评分(TIDS)。测定左肾皮质中超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性、总抗氧化能力(T-AOC)及丙二醛(MDA)含量。蛋白免疫印迹法(Western blot)检测肾皮质中细胞外信号调节激酶1/2(ERK1/2)和磷酸化细胞外信号调节激酶1/2(p-ERK1/2)蛋白表达情况。结果:灌胃给药6 w后,与control组比较,SHR组大鼠尾动脉SBP(2 w、4 w和6 w)、24 h尿蛋白、尿液中RBP和α1-MG含量、BUN、SCr、血清中Cys C含量、肾脏组织中胶原纤维、肾内小动脉MT/LD、GCDS和TIDS、肾皮质中MDA含量、ERK1/2和p-ERK1/2的蛋白表达均明显升高(P0.01,P0.05),而肾皮质中SOD、GSH-Px的活性和T-AOC均明显降低(P0.05,P0.01);肾脏大体形态基本正常,两侧RWI均增大(P0.05),肾小球、肾间质及肾小管均出现损伤性病理改变。与SHR组相比,GSP能显著降低大鼠尾动脉SBP(2 w、4 w和6 w)、24h尿蛋白、尿液中RBP和α1-MG含量、BUN、SCr、血清中Cys C含量(P0.01,P0.05)、肾脏组织中胶原含量、MT/LD、GCDS和TIDS及肾皮质中MDA含量、ERK1/2和p-ERK1/2的蛋白表达(P0.05,P0.01),升高肾皮质中SOD、GSHPx的活性和T-AOC(P0.01),改善肾脏的病理损伤。以高剂量GSP治疗组效果尤为显著(P0.01),与captopril组疗效相当(P0.05)。结论:GSP可以有效改善SHR肾脏结构和泌尿功能的损伤,其机制与GSP具有降低SHR尾动脉SBP、提高SHR抗氧化应激能力和减少肾脏组织中ERK1/2和pERK1/2的蛋白表达的作用有关。
[Abstract]:Aim: to study the effects of grape seed proanthocyanidin (GSP) on renal structure and urinary function in spontaneously hypertensive rats (SHR) and to explore its possible mechanism. Methods Twenty four male SHR (8 weeks old) were randomly divided into 4 groups after one week of adaptive feeding. They were randomly divided into 4 groups: the low dose group of SHRP GSP 50 mg 路kg ~ (-1) GSP) and captopriline 30 mg 路kg ~ (-1) 路kg ~ (-1) of captopriline 30 mg 路kg ~ (-1) 路kg ~ (-1) captopriline 30 mg 路kg ~ (-1) of captopriline 30 mg 路kg ~ (-1 / L) of captoprilium 30 mg 路kg ~ (-1 / L) of captopriline 30 mg 路kg ~ (-1) 路kg ~ (-1) ~ 6 male Wistar Kyoto rats of the same age. According to the experimental group, the rats were gavaged with the corresponding dose of the drug and the solvent for 2 ml/ times / d. The systolic blood pressure of the caudal artery of the rats in each group was measured every 2 weeks. The urine of the rats in each group was collected by metabolic cage for 24 hours, and the urine protein was measured at 24 hours. The contents of Retinal binding protein (RBP) and 伪 1 microglobulin (伪 1 MG). Blood samples were taken from eyeballs and serum samples were collected. The contents of blood urea nitrogen bun, serum creatinine (SCR) and cystatin (cystatin C) were determined. The bilateral kidneys of the rats were removed, the gross morphology of the kidneys was observed with the naked eye, the weight of the rats and the weight of the left and right kidneys were measured. RMI-HE and Masson staining were used to observe the changes of renal tissue morphology and collagen content, and to determine the ratio of membrane to vessel diameter of renal arterioles and the ratio of MTR / LDN, glomerular collagen fiber deposition score (GCDSs) and tubulointerstitial lesion score (TIDSN). The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC) and malondialdehyde (MDA) in left renal cortex were measured. Western blot was used to detect the expression of extracellular signal-regulated kinase 1 / 2 ERK1 / 2 and phosphorylated extracellular signal-regulated kinase 1 / 2 -ERK1 / 2 in renal cortex. Results: after 6 weeks, compared with the control group, the rats in the control group received 24 h urine protein, urinary RBP and 伪 1-MG, Cys C in serum and collagen fiber in the kidney. The expression of MDA in renal cortex, ERK1 / 2 and p-ERK1 / 2 increased significantly, while the activity of SODD-GSH-Px and T-AOC in renal cortex decreased P0.05P0.01. the renal morphology was basically normal, and the bilateral RWI was increased, glomeruli, glomeruli. Interstitial and tubulointerstitial pathological changes were observed. Compared with the SHR group, GSP could significantly decrease the urinary protein levels of SBP in the caudal artery of rats at 4 weeks and 6 weeks, respectively. The contents of RBP and 伪 1-MG in urine, the contents of Cys C in serum, and the contents of collagen in kidney and the protein expression of ERK1 / 2 and p-ERK1 / 2 in renal cortex and the protein expression of ERK1 / 2 and p-ERK1 / 2 in renal cortex were increased, and the activity of GSHPx and T-AOCP0.01in renal cortex were increased, and the pathological damage of kidney was improved. The effect of high dose GSP group was more significant than that of captopril group. ConclusionGSP can effectively improve the damage of renal structure and urinary function in SHR. The mechanism is related to the effect of GSP on decreasing SBP of SHR caudal artery, increasing the ability of anti-oxidative stress in SHR and decreasing the protein expression of ERK1 / 2 and pERK1 / 2 in renal tissue.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R544.1

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