针刺“肾俞”“太溪”穴对高尿酸血症大鼠尿酸的影响及其机制研究
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摘要
目的:通过观察针刺"肾俞""太溪"穴对高尿酸血症大鼠尿酸和肾脏相关尿酸转运蛋白的调控作用,探讨其降尿酸机制。方法:32只雄性Wistar大鼠随机分为4组,剔除死亡大鼠后,最终纳入统计的大鼠为空白组、肝俞组和肾俞组各6只,模型组7只。采用氧嗪酸钾灌胃制备高尿酸血症大鼠模型。肝俞组和肾俞组分别给予针刺"肝俞""太冲"穴和"肾俞""太溪"穴,每日1次,20min/次,连续针刺6次后休息1d,持续3周。全自动生化分析仪检测大鼠血清尿酸(SUA)、肌酐(SCr)含量,HE染色观察大鼠肾脏组织病理学变化,免疫组织化学法和Western blot法分别检测大鼠肾脏尿酸盐阴离子转运体1(URAT1)和有机阴离子转运体1(OAT1)的蛋白表达。结果:与空白组比较,模型组大鼠SUA含量显著升高(P<0.01),肾脏URAT1表达显著升高(P<0.01),OAT1表达显著下降(P<0.01);镜下观察肾脏切片显示,肾小管扩张,炎性细胞浸润,细胞空化严重。与模型组比较,肝俞组和肾俞组SUA含量均显著下降(P<0.05,P<0.01),URAT1表达下降(P<0.05,P<0.01),肾俞组OAT1表达显著升高(P<0.01)。与肝俞组比较,肾俞组URAT1表达显著下降(P<0.01),OAT1表达显著升高(P<0.05)。与模型组比较,肝俞组和肾俞组肾小管扩张程度减轻,细胞空化现象减少,而肾俞组较肝俞组肾脏损伤程度更轻。结论:针刺"肾俞""太溪"穴可显著降低大鼠SUA含量,其机制可能与抑制大鼠肾脏URAT1表达而减少了尿酸重吸收,增加了肾脏OAT1表达有关。
Objective To observe the effect of acupuncture at"Shenshu"(BL23)-"Taixi"(KI3)on the levels of serum uric acid(SUA)and expression of renal urate-anion transporter 1(URAT1)and organic anion transporter 1(OAT1)proteins in hyperuricemia(HUA)rats,so as to explore its underlying mechanisms in improving HUA.Methods A total of 25 male Wistar rats were divided into 4 groups:control(n=6),HUA model(n=7),BL23-KI3(n=6)and Ganshu(BL18)-Taichong(LR3,BL18-LR3 in short,n=6).The HUA model was established by gavage of Oteracil Potassium(2 g/kg),once daily for 10 days,then once every other day.For rats of the BL23-KI3 group,BL23 and KI3 were stimulated with filiform needles which were rotated for 10 sat a frequency about 100 r/min,and for rats of the BL18-LR3 group,BL18 and LR3 were stimulated with the same methods to those of the BL23-KI3 group.The treatment of both acupuncture groups was conducted once daily,6 times a week(except Sundays)for 3 weeks.The contents of SUA and serum creatinine(SCr)were assayed by using an automatic biochemical analyzer.The pathological changes of the right kidney tissue were observed under light microscope after hematoxylin eosin(H.E.)staining,the immunoactivity of URAT1 and OAT1 of the right kidney tissue was determined by immunohistochemistry,and the expression of URAT1 and OAT1 proteins of the left kidney tissue detected by Western blot(WB).Results After modeling,the content of SUA and the expression of renal URAT1 protein(shown by both immunoactivity and WB)were significantly increased(P<0.01),but that of renal OAT1 protein was obviously decreased in the model group compared with the control group(P<0.01).There was no notably change in the level of SCr in the model group relevant to the control group(P>0.05).Following acupuncture intervention,the SUA content and URAT1 expression in both BL18-LR3 and BL23-KI3 groups were considerably down-regulated(P<0.05,P<0.01),and the expression of OAT1 protein in the BL23-KI3 group(not the BL18-LR3 group)were obviously up-regulated relevant to the model group(P<0.01).The effects of BL23-KI3 were significant superior to those of BL18-LR3 in down-regulating the expression of URAT1 and up-regulating OAT1 protein(P<0.01,P<0.05).Conclusion Acupuncture of"BL23"and"KI3"can effectively down-regulate SUA level in HUA rats,which may be related to its effects in down-regulating the expression of URAT1 and up-regulating the expression of OAT1 in the kidney tissue.
Objective To observe the effect of acupuncture at"Shenshu"(BL23)-"Taixi"(KI3)on the levels of serum uric acid(SUA)and expression of renal urate-anion transporter 1(URAT1)and organic anion transporter 1(OAT1)proteins in hyperuricemia(HUA)rats,so as to explore its underlying mechanisms in improving HUA.Methods A total of 25 male Wistar rats were divided into 4 groups:control(n=6),HUA model(n=7),BL23-KI3(n=6)and Ganshu(BL18)-Taichong(LR3,BL18-LR3 in short,n=6).The HUA model was established by gavage of Oteracil Potassium(2 g/kg),once daily for 10 days,then once every other day.For rats of the BL23-KI3 group,BL23 and KI3 were stimulated with filiform needles which were rotated for 10 sat a frequency about 100 r/min,and for rats of the BL18-LR3 group,BL18 and LR3 were stimulated with the same methods to those of the BL23-KI3 group.The treatment of both acupuncture groups was conducted once daily,6 times a week(except Sundays)for 3 weeks.The contents of SUA and serum creatinine(SCr)were assayed by using an automatic biochemical analyzer.The pathological changes of the right kidney tissue were observed under light microscope after hematoxylin eosin(H.E.)staining,the immunoactivity of URAT1 and OAT1 of the right kidney tissue was determined by immunohistochemistry,and the expression of URAT1 and OAT1 proteins of the left kidney tissue detected by Western blot(WB).Results After modeling,the content of SUA and the expression of renal URAT1 protein(shown by both immunoactivity and WB)were significantly increased(P<0.01),but that of renal OAT1 protein was obviously decreased in the model group compared with the control group(P<0.01).There was no notably change in the level of SCr in the model group relevant to the control group(P>0.05).Following acupuncture intervention,the SUA content and URAT1 expression in both BL18-LR3 and BL23-KI3 groups were considerably down-regulated(P<0.05,P<0.01),and the expression of OAT1 protein in the BL23-KI3 group(not the BL18-LR3 group)were obviously up-regulated relevant to the model group(P<0.01).The effects of BL23-KI3 were significant superior to those of BL18-LR3 in down-regulating the expression of URAT1 and up-regulating OAT1 protein(P<0.01,P<0.05).Conclusion Acupuncture of"BL23"and"KI3"can effectively down-regulate SUA level in HUA rats,which may be related to its effects in down-regulating the expression of URAT1 and up-regulating the expression of OAT1 in the kidney tissue.
引文
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[5]高尿酸血症相关疾病诊疗多学科共识专家组.中国高尿酸血症相关疾病诊疗多学科专家共识[J].中华内科杂志,2017,56(3):235-249.
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[8]李柯,于彩娜.济生肾气丸对高尿酸血症大鼠的影响及作用机制[J].中华中医药学刊,2017,35(7):1882-1885.
[9]高小涛,高俊玲,佟晓波.大鼠高尿酸血症模型构造概况[J].临床医药文献电子杂志,2017,4(11):1996.
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[12]嘉士健,雷行华,廖建坤.针灸结合中药治疗高尿酸血症48例[J].实用中医药杂志,2014,30(10):935.
[13]陆欣玲,李瑞玲,沈卫东,等.针刺治疗高尿酸血症的临床效果[J].中国医药导报,2017,14(25):102-105.
[14]杨雪,王薇,张传宝,等.我国肌酐检测系统性能分析[J].检验医学,2012,27(12):989-994.
[15]姚来隽.血尿酸检测指导诊断高尿酸血症及痛风的观察[J].内蒙古中医药,2016,35(3):96-97.
[16]杨桂梅,黄胜华,李江,等.腺嘌呤和氧嗪酸制作高尿酸血症大鼠模型比较[J].实验动物科学,2011,28(2):23-26.
[17] ENOMOTO A,KIMURA H,CHAIROUNGDUA A.et al.Molecular identification of a renal urate-anion exchanger that regulates blood urate levels[J].Nature,2002,417(6887):447-452.
[18]吕森森.hURAT1基因3’非编码区基因变异筛查及与原发性高尿酸血症关联性研究[D].青岛:青岛大学,2009:10.
[19] SHEIKH M, MOVASSAGHI S, KHALEDI M,et al.Hyperuricemia in systemic lupus erythematosus:is it associated with the neuropsychiatric manifestations of the disease[J].Rev Bras De Reumatol,2016,56(6):471-477.
[20] ERALY S A,VALLON V,RIEG T,et al.Multiple organic anion transporters contribute to net renal excretion of uric acid[J].Physiol Genomics,2008,33(2):180-192.
[21]刘孟渊.中药干预尿酸转运蛋白及基因表达的研究进展[J].上海中医药杂志,2018,52(8):93-97.
[2] KUWABARA M,HISATOME I,NIWA K,et al,Uric acid is a strong risk marker for developing hypertension from prehypertension:a 5-year Japanese cohort study[J].Hypertension,2018,71(1):78-86.
[3] BORGHI C,RODRIGUEZ-ARTALEJO F,DE BACKER G,et al.Serum uric acid levels are associated with cardiovascular risk score:apost hoc analysis of the EURIKA study[J].Intern J Cardiol,2018,25(3):167-173.
[4]赵卫云,张慧敏,李辉,等,高尿酸血症与代谢综合征及其组分的相关性[J].心脏杂志,2016,28(4):457-459.
[5]高尿酸血症相关疾病诊疗多学科共识专家组.中国高尿酸血症相关疾病诊疗多学科专家共识[J].中华内科杂志,2017,56(3):235-249.
[6] RICHES P L,WRIGHT A F,RALSTON S H.Recent insights into the pathogenesis of hyperuricaemia and gout[J].Hum Mol Genet,2009,18(R2):R177-R184.
[7]王心怡.针刺俞、原、募穴对高尿酸血症大鼠SUA、XOD、ALP的影响[D].北京:北京中医药大学,2017.
[8]李柯,于彩娜.济生肾气丸对高尿酸血症大鼠的影响及作用机制[J].中华中医药学刊,2017,35(7):1882-1885.
[9]高小涛,高俊玲,佟晓波.大鼠高尿酸血症模型构造概况[J].临床医药文献电子杂志,2017,4(11):1996.
[10]于曙光,徐斌.实验针灸学[M].北京:中国中医药出版社,2013.
[11] HU Q H,JIAO R Q,WANG X,et al.Simiao pill ameliorates urate underexcretion and renal dysfunction in hyperuricemic mice[J].J Ethnopharmacol,2010,128(3):685-692.
[12]嘉士健,雷行华,廖建坤.针灸结合中药治疗高尿酸血症48例[J].实用中医药杂志,2014,30(10):935.
[13]陆欣玲,李瑞玲,沈卫东,等.针刺治疗高尿酸血症的临床效果[J].中国医药导报,2017,14(25):102-105.
[14]杨雪,王薇,张传宝,等.我国肌酐检测系统性能分析[J].检验医学,2012,27(12):989-994.
[15]姚来隽.血尿酸检测指导诊断高尿酸血症及痛风的观察[J].内蒙古中医药,2016,35(3):96-97.
[16]杨桂梅,黄胜华,李江,等.腺嘌呤和氧嗪酸制作高尿酸血症大鼠模型比较[J].实验动物科学,2011,28(2):23-26.
[17] ENOMOTO A,KIMURA H,CHAIROUNGDUA A.et al.Molecular identification of a renal urate-anion exchanger that regulates blood urate levels[J].Nature,2002,417(6887):447-452.
[18]吕森森.hURAT1基因3’非编码区基因变异筛查及与原发性高尿酸血症关联性研究[D].青岛:青岛大学,2009:10.
[19] SHEIKH M, MOVASSAGHI S, KHALEDI M,et al.Hyperuricemia in systemic lupus erythematosus:is it associated with the neuropsychiatric manifestations of the disease[J].Rev Bras De Reumatol,2016,56(6):471-477.
[20] ERALY S A,VALLON V,RIEG T,et al.Multiple organic anion transporters contribute to net renal excretion of uric acid[J].Physiol Genomics,2008,33(2):180-192.
[21]刘孟渊.中药干预尿酸转运蛋白及基因表达的研究进展[J].上海中医药杂志,2018,52(8):93-97.