高盐饮食对Wistar大鼠肾损害的影响及替米沙坦和辣椒素干预

发布时间：2018-07-14 16:57

【摘要】：目的：探讨高盐饮食对Wistar大鼠肾损害的影响及替米沙坦和辣椒素的干预作用。方法：Wistar雄性大鼠65只，随机分为正常盐对照组（NSD组，0.5%NaCl的颗粒饲料喂养），高盐组（HSD组，4%NaCl的颗粒饲料喂养），高盐+替米沙坦组（HSD+ARB组，4%NaCl的颗粒饲料喂养+替米沙坦），高盐+辣椒素组（HSD+CAP组，，4%NaCl的颗粒饲料+辣椒素），每两周测定尾动脉压一次，喂养24周。实验结束后，根据尾动脉压再将高盐组分为高盐高血压组（HSD-H组）、高盐正常血压组（HSD-N组）。各组大鼠取双肾， HE染色观察肾脏的形态结构，Masson染色观察肾脏及血管壁的纤维化程度；阶段性收集24小时大鼠尿液测定Na+、K+、微量白蛋白、视黄醇结合蛋白（RBP）、TH糖蛋白（THP）、末次尿总蛋白及肌酐，评价肾功能；检测肾脏皮质Na+-K+-ATP酶、Ca2+-ATP酶活性；免疫组化法检测Na+-K+-ATPaseα1、PPARγ、p-NF-κB p65的表达；运用real time RT-PCR法检测肾皮质Na+-K+-ATPaseα1、PMCA1的mRNA表达；免疫印迹法检测皮质Na+-K+-ATPaseα1、皮髓质PPARγ、皮髓质p-NF-κB p65的蛋白表达。结果：与对照组相比，高盐高血压组较其他各组血压升高（P0.05）。HE和Masson结果显示高盐组肾脏形态结构有不同程度的损害，替米沙坦和辣椒素可明显改善其肾损害。与对照组相比，高盐组、替米沙坦组和辣椒素组24h尿钠钾比值、24小时尿总蛋白增高（P0.05）；与对照组相比，高盐组24小时尿微量白蛋白显著增高（P0.05），替米沙坦组和辣椒素组低于高盐组（P0.05），高盐高血压组24小时尿微量白蛋白高于高盐正常血压组（P0.05）；与对照组相比，高盐组、替米沙坦组和辣椒素组24h尿视黄醇结合蛋白和TH糖蛋白升高（P0.05），高盐组和替米沙坦组肌酐清除率降低（P0.05），而辣椒素组肌酐清除率无改变。高盐组肾皮质Na+-K+-ATP酶活性和Ca2+-ATP酶活性都明显低于对照组（P0.05），而Na+-K+-ATPaseα1、PMCA1的mRNA表达增高。与对照组相比，高盐组、替米沙坦组肾脏皮髓质PPARγ、p-NF-κB p65表达明显增高（P0.05）。结论：（1）长期高盐饮食可导致部分Wistar大鼠血压升高，部分大鼠血压不升高。（2）高盐饮食可独立于血压直接导致肾功能损害。（3）高盐饮食致Wistar大鼠肾功能损伤的机制可能与钠、钙泵活性降低及PPARγ、NF-κB p65激活有关。（4）替米沙坦具有降压和改善肾功能的作用，可能与激活PPARγ及抑制NF-κB p65蛋白表达有关。（5）辣椒素具有降压和改善肾损害的作用。
[Abstract]:Aim: to investigate the effects of high salt diet on renal damage in Wistar rats and the effects of telmisartan and capsaicin. Methods Sixty-five male Wistar rats were used. They were randomly divided into normal salt control group (NSD group), high salt group (HSD group), high salt telmisartan group (HSD ARB group), high salt capsaicin group (HSD CAP group), high salt telmisartan group (HSD ARB group) and high salt capsaicin group (HSD CAP group). The caudal arterial pressure was measured every two weeks. Feeding for 24 weeks. After the experiment, according to the caudal arterial pressure, the high salt components were divided into high salt hypertension group (HSD-H group) and hypersalt normal blood pressure group (HSD-N group). The two kidneys were taken from each group, the morphology and structure of kidney were observed by HE staining and the degree of fibrosis of kidney and vascular wall was observed by Masson staining. Retinol binding protein (RBP) TH glycoprotein (THP), total protein and creatinine in the last urine were used to evaluate renal function, the activity of Na K ATPase Ca 2 + ATPase in renal cortex and the expression of Na K ATPase 伪 1 PPAR 纬 -nf 魏 B p65 in renal cortex were detected by immunohistochemistry. Real time RT-PCR was used to detect the expression of Na -K -ATPase 伪 1 and protein expression of p-NF- 魏 B p65 in cortical Na K ATPase 伪 1, skin medulla PPAR 纬 and medulla p65 in renal cortex. Results: compared with the control group, the blood pressure in the high salt hypertension group was higher than that in the other groups (P0.05). The results of HE and Masson showed that the renal morphology in the high salt group was damaged to some extent, and telmisartan and capsaicin could significantly improve the renal damage. Compared with control group, the ratio of 24 hours urinary sodium and potassium in high salt group, telmisartan group and capsaicin group was higher than that in control group (P0.05). In high salt group, 24 hours urinary microalbumin was significantly increased (P0.05), telmisartan group and capsaicin group were lower than those in high salt group (P0.05), 24 hour urinary microalbumin in high salt hypertension group was higher than that in high salt normal blood pressure group (P0.05), compared with control group, high salt group, In telmisartan group and capsaicin group, 24 hours urinary retinol binding protein and th glycoprotein were increased (P0.05), creatinine clearance was decreased in high salt group and telmisartan group (P0.05), but creatinine clearance rate was not changed in capsaicin group. The Na K ATPase activity and Ca 2 ATPase activity in the renal cortex of high salt group were significantly lower than those in the control group (P 0.05), while the mRNA expression of Na K ATPase 伪 1 and PMCA 1 was increased. Compared with the control group, the expression of PPAR 纬 -NF- 魏 B p65 in renal medulla was significantly increased in high-salt group and telmisartan group (P0.05). Conclusion: (1) Long-term high salt diet can increase blood pressure of some Wistar rats, while some rats' blood pressure does not. (2) High salt diet can directly lead to renal function damage independently of blood pressure. (3) the mechanism of renal function damage in Wistar rats induced by high salt diet may be related to sodium. The decrease of calcium pump activity and the activation of PPAR 纬 -NF- 魏 B p65. (4) telmisartan can reduce blood pressure and improve renal function, which may be related to activation of PPAR 纬 and inhibition of NF- 魏 B p65 protein expression. (5) capsaicin can reduce blood pressure and ameliorate renal damage.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R544.1;R692

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