【作者】 张晓华；

【导师】 隋殿军；

【作者基本信息】 长春中医药大学 ， 中医内科学， 2013， 博士

【摘要】 心力衰竭(简称心衰)属心血管疾病的急危重证,严重威胁人类健康,因心衰病人多伴有水肿、胸水、腹水等,故心衰也属水液代谢障碍性疾病,对于心衰水液潴留机制的研究,很多学者从多个系统进行过研究,包括肾素-血管紧张素-醛固酮系统(RAAS)、精氨酸加压素系统、交感神经系统、内皮素系统、利钠肽系统(心房利钠肽、脑利钠肽)等等,随着水通道蛋白的发现,尤其是水通道蛋白2的出现,为研究心力衰竭水液潴留机制开辟了一个新的途径,精氨酸血管加压素-血管加压素V2受体-水通道蛋白2系统在心衰水液潴留过程中的作用逐渐被学者们重视及研究,现有学者指出心力衰竭水潴留机制的主要途径为心输血量下降-血管加压素水平升高-肾脏集合管V2受体激活-水通道蛋白2激活和／或基因表达上调-水重吸收增加,水通道蛋白2(AQP2)主要受精氨酸血管加压素(AVP)调控,而AVP通过肾脏V2受体调控AQP2,故AVP-V2受体-AQP2系统在调节机体水液代谢中起着重要作用。益气活血,温阳利水法是目前公认的治疗心衰的大法,是根据心衰的病机：气阳亏虚,血瘀水结而定的,目前中医药对于本法的实验研究主要集中在血流动力学、RAAS系统、利钠肽系统、心肌重塑等方面,对于水潴留机制的研究还处于起步阶段,现有研究主要是单味中药(如茯苓、泽泻、大黄、黄芪、丝瓜络等)对于水通道蛋白2的影响,也有部分学者应用中药复方观察其对水通道蛋白2的影响,而对于整个精氨酸血管加压素-V2受体-水通道蛋白2系统的研究较少,未能从整体上阐明本法对于心衰水潴留的作用机制。目的：研究益气活血温阳利水法对于心衰大鼠精氨酸血管加压素-V2受体-水通道蛋白2系统的影响,以探讨本法对心衰水液潴留的作用机理。方法：选择健康Wistar大鼠,应用腹腔注射阿霉素(3mg/kg,每周1次,连续6周)及睡眠剥夺,造成心衰大鼠模型,然后分空白组、模型组,芪苈强心组、中药高剂量、中剂量、低剂量6组,服药4周后应用ELISA法观察6组大鼠服药后血浆精氨酸血管加压素(AVP)浓度；应用免疫组化、蛋白印迹(Western-Blot)方法检测肾组织(近髓部)血管加压素V2受体(V2R)、水通道蛋白2(AQP2)蛋白表达情况；应用rt-PCR检测肾组织(近髓部)血管加压素V2受体、水通道蛋白2mRNA相对表达量的变化情况。结果：(?)较,有统计学差异；可减弱肾脏组织V2R、AQP2蛋白表达及m RNA相对表达量,结果优于模型组,以中、高剂量组为著。结论：益气活血温阳利水法通过降低血浆中AVP浓度,下调V2R、AQP2蛋白表达、m RNA因子表达来作用于精氨酸血管加压素-V2受体-水通道蛋白2系统的每个环节而起到改善水液潴留之目的。更多还原

【Abstract】 Heart failure (HF) is critical symptom in cardiovascular disease and threatens people’s life. HF is water metabolic obstacle disease with edema, thorax edema and abdomen edema. Scholars study on HF water retention mechanism from many systems including rennin angiotensin aldosternone system (RAAS), arginine vasopressin (AVP), sympathetic nerves system,endothelin system, natriuretic peptide system (ANP and BNP), etc. With the discovery of aquaporin, especially aquaporin2, new research on HFwater retention system has been developed. The effect of arginine vasopressin (AVP)-V2RA-AQP2system in HF water retention process has been gradually noticed and studied. Some scholars think that HF water retention system results from the decrease of heart blood transfusion-the increase of arginine vasopressin level-renal receptor antagon-the activation of aquaporin2and/the increase of gene level-the increase of water reabsorption; and aquaporin2is mainly controlled by AVP, but AVP manipulates AQP2through renal receptor antagon, so AVP-V2receptor-AQP2system takes very important effect in water metabolism.The method of yiqihuoxuewenyanglishui has been generally accepted as the main approach these days according to qiyangkuixu and xueyushuijie. Recently, traditional Chinese medicine and pharmacy focuses on hemodynamics, RAAS, natriuretic peptide and myocardial remodeling, etc. The study on water retention mechanism is in initial stage. Research has just been found in the effect of signal Chinese material medica (such as fuling, zexie, dahuang, huangqi and sigualuo and so on) on AQP2, and compound Chinese material medica can also affect on AQP2. However, with few research on V2R-AQP2, the effect mechanism of yiqiwenyanghuoxuelishui method could not been expounded.Objective:To study the effect of yiqihuoxuewenyanglishui method on the rat of heart failure AVP-V2R-AQP2system in order to explore the effect on HF water retention mechanism by means of this method.Method:Healthy Wistar rats have been chosen, which have been injected adriamycin through abdominal cavity (3mg/kg, once a week, succession of6weeks) with sleep deprivation to make model of HF rats. Then six groups could be divided:blank, model, liqi cardiac tonic, high, medium and low dose Chinese medicine. Four weeks later, six groups of rats with medicine could be observed the concentration of plasma AVP with the method of ELISA, and could be tested the condition of protease of renal organization (spinal cord) V2R, AQP2with immunohistochemical and western-blot; could also be tested relative variety of2m in RNA renal organization (spinal cord) V2R, AQP2with the application of rt-PCR. Result:Method of Yiqihuoxuewenyanglishui could decrease concentration of HF rats’high AVP concentration. Compared with group model, statistical difference could be found. The method could also weaken renal organization V2R, AQP2protease and m RNA relative quantity; so that the result was better than group model especially group medium and high dose.Conclusion:Method of yiqihuoxuewenyanglishui decreases the AVP concentration of plasma and V2R, AQP2protease, mRNA factor to take effect on each section in V2R-AQP2system, and then improve water retention.更多还原