大鼠Klotho蛋白水平随血压升高而降低  

Klotho Protein Levels in Rats Decreased with Increased Blood Pressure

作　　者：王彦琛 杨伟[2] 张玮[2] 罗丹 苏显明[2] WANG Yan-Chen;YANG Wei;ZHANG Wei;LUO Dan;SU Xian-Ming(Department of Geriatrics,Xianyang Central Hospital,Xianyang 712000,China;Department of Geriatric Cardiology,First Affiliated Hospital of Xi'an Jiao-Tong University,Xi'an 710061,China)

机构地区：[1]咸阳市中心医院老年病科,陕西咸阳712000 [2]西安交通大学第一附属医院老年心血管内科,陕西西安710061

出　　处：《生物技术通讯》2020年第4期432-438,共7页Letters in Biotechnology

基　　金：陕西省自然科学基金(2016JM8082)。

摘　　要：目的:探讨Klotho(kl)蛋白在大鼠高血压发生发展过程中的变化。方法:用不同浓度的N'-硝基-L-精氨酸干预大鼠肾小管上皮细胞,PCR检测kl基因表达的变化。选取48只体质量200±10 g的7周龄雄性SD大鼠,随机分为高血压组和对照组。高血压组大鼠造模用N'-硝基-L-精氨酸98 mg/d连续灌胃,对照组用等体积生理盐水灌胃,共28 d,2组大鼠分别于停药后第7、14、21、28 d测体质量、尾动脉血压,每组抽取6只体质量相近的大鼠用水合氯醛麻醉后腹主动脉采血,ELASA检测血浆中的kl蛋白、血栓素(TXA2)、内皮素(ET)水平,硝酸还原酶法检测一氧化氮(NO)水平,TBA法检测丙二醛(MDA)含量,WST-1法检测超氧化物歧化酶(SOD)活力。观察停药后第7、14、21、28 d胸主动脉HE染色血管壁病理变化,免疫组化CD31标记观察血管内皮细胞损伤情况。结果:在α=0.05水准上,2组大鼠收缩压在各因素影响下差异有统计学意义(P<0.05)。校正模型检验时各指标的P值均<0.05,说明模型均有统计学意义。血压、时间之间的交互作用对kl蛋白、NO浓度的影响有统计学意义(P<0.05)。HE染色结果显示,高血压组随着高血压的发生主动脉壁弹力纤维增生、排列紊乱,中膜逐渐增厚。免疫组化CD31标记结果显示,高血压组随着高血压的发生内皮细胞受损,逐渐减少、脱落。结论:kl蛋白、NO水平随血压升高逐渐降低,与血压呈负相关;kl蛋白浓度与NO水平变化呈相同趋势,推测kl蛋白的减少通过某种机制使血管内皮分泌NO减少,从而介导了大鼠高血压的发生发展。Objective:To investigate the relationship between klotho(kl)protein in the development of hypertension in rats.Methods:Different concentrations of N’-nitro-L-arginine were used to intervene in renal tubular epithelial cells in rats,and the changes of klotho protein were detected by PCR.Forty-eight male SD rats aged 7 weeks were selected with normal blood pressure were randomly divided into hypertensive group and control group with 24 rats in each group.The hypertension rat model was established in hypertensive group by intragastric administration of N’-nitro-L-arginine methyl ester in drinking water(98 mg/d),and each group measured indicators by the time point(7,14,21 and 28 days)after discontinuation as follows:rat body quality,tail artery blood pressure,blood biochemical index(the level or activity of plasma klotho protein,thromboxane,endothelin,nitric oxide,malondialdehyde,and superoxide dismutase).The pathological changes of thoracic aorta by HE staining,and the damage of vascular endothelial cells by immunohistochemical CD31 markers.Results:At the level of α=0.05,the difference of systolic blood pressure between the two groups was statistically significant under the influence of various factors(P<0.05).P values of all indexes in the calibration model test were all<0.05,all the models were statistically significant.The interaction between blood pressure and time had a statistically significant effect on kl protein concentration and nitric oxide concentration(P<0.05).The results of HE staining showed that the hypertensive group was thickened with the hyperplasia and disorder of the aortic wall elastic fibers.Immunohistochemical CD31 marker results showed that the endothelial cells in the hypertensive group were damaged,gradually decreased and dropped.Conclusion:Klotho protein,nitric oxide level with the decreasing blood pressure increases gradually,negatively correlated with blood pressure.It is speculated that the decrease of kl protein causes the decrease of nitric oxide secretion in vascular endotheliu

关 键 词：高血压 KLOTHO蛋白 发病机制 大鼠 

分 类 号：R544.1[医药卫生—心血管疾病]