针刺对心衰模型大鼠血浆中精氨酸加压素浓度及水通道蛋白2表达的影响
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摘要
目的观察针刺对心衰模型大鼠血浆中精氨酸加压素(AVP)浓度及水通道蛋白2(AQP2)表达的影响,并探讨其可能的调节机制。方法将大鼠随机分为空白组、模型组及针刺组。除外空白组,其余各组大鼠采用腹腔注射阿霉素(3 mg/kg,每周1次,连续6周)及睡眠剥夺复制心衰模型。空白组与模型组采取与针刺组相同的固定,针刺组予针刺双侧心俞穴、双侧肾俞穴、双侧内关穴治疗。均1次/d,连续治疗15 d。干预结束后,观察大鼠血浆中精氨酸加压素(AVP)浓度变化;采用免疫组化、蛋白印迹方法检测水通道蛋白2(AQP2)蛋白表达;实时定量PCR检测水通道蛋白2(AQP2)在大鼠肾组织的表达量的变化情况。结果针刺治疗组与未治疗的模型组相比,AVP浓度明显降低,并具有统计学意义(P <0.01),针刺治疗组血浆中AVP浓度低于对照组,但无统计学意义(P> 0.05);蛋白印迹结果显示,各组灰度值进行对比,针刺组与模型组相比,差异有统计学意义(P <0.01),模型组与空白组相比,差异有统计学意义(P <0.05);模型组心衰大鼠AQP2-m RNA表达明显升高,与对照组比较,差异有统计学意义(P <0.01)。针刺治疗组AQP2-m RNA明显下降与模型组比较,差异有统计学意义(P <0.01)。结论针刺可减轻大鼠心力衰竭症状,其作用机制可能与下调AVP浓度及AQP2表达而改善心力衰竭水潴留状态有关。
Objective To observe the effect of acupuncture on the concentration of arginine vasopressin(AVP) and the expression of aquaporin 2(AQP2) in the plasma of rats with heart failure, and to explore the possible regulatory mechanism. Methods Rats were randomly divided into blank group, model group and acupuncture group. Except for the blank group, the other groups were given intraperitoneal injection of doxorubicin(3 mg/kg, 1 time/w, continuous6 weeks) and sleep deprivation to replicate the heart failure model. The blank group and the model group were treated with the same fi xation as the acupuncture group. The acupuncture group was treated with bilateral Xinshu, bilateral Shenshu and bilateral Neiguan points. The treatment group was treated 1 time a day for a continuous treatment of15 d. After the intervention, the concentration of arginine vasopressin(AVP) in the rat plasma was observed and theexpression of water channel protein 2(AQP2) protein was detected by immunohistochemistry and Western blotting,and the changes of the expression of aquaporin 2(AQP2) in the renal tissue of rats were detected by real-time quantitative PCR. Results Compared with the untreated model group, the concentration of AVP was significantly lower than that in the untreated group(P < 0.01). The concentration of AVP in the plasma of the acupuncture treatment group was lower than that of the control group, but there was no signifi cant difference(P > 0.05). The results of Western blotting showed that the gray value of each group was compared. There was signifi cant difference between the acupuncture group and the model group(P < 0.01), and the model group was statistically signifi cant compared with the blank group(P < 0.05). The expression of AQP2-mRNA in the model group was signifi cantly higher than that in the control group, and there was a signifi cant difference between the model group and the control group(P < 0.01). The AQP2-mRNA of acupuncture treatment group decreased signifi cantly compared with the model group P <0.01, there was a signifi cant difference. Conclusion Acupuncture can relieve heart failure in rats, and its mechanism may be related to the reduction of AVP concentration and AQP2 expression to improve the state of water retention in patients with heart failure.
Objective To observe the effect of acupuncture on the concentration of arginine vasopressin(AVP) and the expression of aquaporin 2(AQP2) in the plasma of rats with heart failure, and to explore the possible regulatory mechanism. Methods Rats were randomly divided into blank group, model group and acupuncture group. Except for the blank group, the other groups were given intraperitoneal injection of doxorubicin(3 mg/kg, 1 time/w, continuous6 weeks) and sleep deprivation to replicate the heart failure model. The blank group and the model group were treated with the same fi xation as the acupuncture group. The acupuncture group was treated with bilateral Xinshu, bilateral Shenshu and bilateral Neiguan points. The treatment group was treated 1 time a day for a continuous treatment of15 d. After the intervention, the concentration of arginine vasopressin(AVP) in the rat plasma was observed and theexpression of water channel protein 2(AQP2) protein was detected by immunohistochemistry and Western blotting,and the changes of the expression of aquaporin 2(AQP2) in the renal tissue of rats were detected by real-time quantitative PCR. Results Compared with the untreated model group, the concentration of AVP was significantly lower than that in the untreated group(P < 0.01). The concentration of AVP in the plasma of the acupuncture treatment group was lower than that of the control group, but there was no signifi cant difference(P > 0.05). The results of Western blotting showed that the gray value of each group was compared. There was signifi cant difference between the acupuncture group and the model group(P < 0.01), and the model group was statistically signifi cant compared with the blank group(P < 0.05). The expression of AQP2-mRNA in the model group was signifi cantly higher than that in the control group, and there was a signifi cant difference between the model group and the control group(P < 0.01). The AQP2-mRNA of acupuncture treatment group decreased signifi cantly compared with the model group P <0.01, there was a signifi cant difference. Conclusion Acupuncture can relieve heart failure in rats, and its mechanism may be related to the reduction of AVP concentration and AQP2 expression to improve the state of water retention in patients with heart failure.
引文
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[2]张秋梅,孙增玉,张艳,等.参草通脉颗粒影响慢性心衰大鼠心肌重构的作用机制研究[J].中华中医药学刊, 2018,36(4):808-811.
[3]郭长升,赵京生,王香荣,等.心得宁口服液对大鼠慢性心衰模型左心室指数,全心指数及心脏动力学的影响[J].中华中医药学刊, 2018, 36(3):672-674.
[4]李忠仁.实验针灸学[M].北京:中国中医药出版社, 2003.
[5]HARTFORDM, WIKLUNDO,MATTSSONHL,etal.C-reactive protein, interleukin-6, secretory phospholipase A 2group II A and intercellular adhesion molecule-1 in the prediction of late outcome events after acute coronary syndromes[J]. Journal of Internal Medicine, 2007, 262(5):526-536.
[6]徐陵琦,陈慧娟.急、慢性抑制CaMKⅡ的活性对心衰小鼠心功能和心力储备的影响[J].中国临床研究, 2018,31(2):158-162.
[7]张晓华,刘淑荣,钱锋,等.强心康颗粒对慢性心衰大鼠心肌细胞病理形态学腺苷酸转位酶, PGC-1αmRNA表达的影响[J].中国实验方剂学杂志, 2018, 24(9):121-126.
[8]李思耐,林谦,高群,等.黄芪党参对心肌梗死后心衰小鼠心肌复极时间的干预研究[J].中国中西医结合杂志, 2018,38(2):232-236.
[9]马笑凡,王守富,崔伟锋,等.从痰湿(瘀)辨治慢性心衰[J].实用中医内科杂志, 2017, 31(11):1-2.
[10]孙刚,张建玲,王治义,等.心衰宁治疗慢性心力衰竭心肾阳虚型临床研究[J].时珍国医国药, 2017, 28(11):2686.
[11]KATO T,NIIZUMAS,INUZUKA Y,etal. Analysisof metabolicremodelingincompensatedleftventricular hypertrophy and heart failure[J]. Circulation-Heart Failure,2010, 3(3):420-430.
[12]杨玥欣,沈夏波,武雅静,等.高脂血症大鼠下颌下腺水通道蛋白2和4的表达[J].中国老年学杂志, 2018,38(6):1482-1484.
[13]沈明.Obestatin对心力衰竭大鼠肾脏水通道蛋白2短时调控作用的研究[D].上海:第二军医大学, 2017.
[14]方庆.当归芍药散对肾病综合征大鼠一氧化氮和水通道蛋白2表达的影响及机制研究[D].合肥:安徽中医药大学, 2017.
[15]王艳霞,张可,礼海,等.心衰参七强心方联合西药对慢性心衰患者血清脑利钠肽及左室射血分数的影响[J].长春中医药大学学报, 2014, 30(6):1082-1084.