电针对高血压前期Dahl大鼠肾脏CaM/NO通路的影响

摘要

目的 探讨电针对高血压前期Dahl盐敏感大鼠模型肾脏CaM-eNOS-NO信号通路的影响.方法 将30只7周龄雄性Dahl盐敏感大鼠(Dahl salt-sensitive rats,DS)按照随机数字表法随机分为模型组、针刺组和非经非穴组.选择7周龄雄性盐抵抗大鼠(Dahl salt-resistant rats,DR) 10只作为空白组.大鼠统一喂养于SPF动物实验中心,在1星期适应性喂养结束后,改为8％ NaC1高盐饲料喂食.其间采用智能无创血压计监测大鼠血压,当12d后血压值升至120～139/80～89mmHg范围时,确定为高血压前期大鼠模型.停止高盐喂养,改为普通饲料.空白组、模型组采取普通固定15 min,不予治疗;针刺组予以双侧曲池、足三里电针治疗;非经非穴组在双侧髂嵴上i0～15 mm、后正中线旁开20 mm区段内选择1个固定对照点进行电针治疗.每日1次,每星期治疗6次,连续治疗5星期.取大鼠左肾组织,荧光定量PCR法检测CaM、eNOS的mRNA表达,western blot 和免疫组化法对eNOS进行定量和定位检测,Elisa法测定NO含量表达.结果 与空白组比较,模型组CaM、eNOS的mRNA表达显著降低(P＜0.05),eNOS的蛋白含量明显减少(P＜0.05),NO含量也明显降低(P＜0.05).与模型组比较,针刺组CaM、eNOS的mRNA表达显著升高(P＜0.05),eNOS的蛋白含量显著增多(P＜0.05),NO含量也明显升高(P＜0.05).与模型组比较,非经非穴组CaM表达虽明显增高(P＜0.05),但是eNOS中mRNA表达、NO含量均无统计学差异(P＞0.05).结论 电针曲池、足三里对高血压前期DS大鼠肾脏保护作用可能与上调大鼠肾脏CaM-eNOS-NO信号通路有关.%Objective To investigate the effect of electroacupuncture on renal CaM-eNOS-NO signal pathway in Dahl salt-sensitive rats with prehypertension.Methods Thirty 7-week-old male Dahl salt-sensitive rats (DS) were allocated,using a random number table,to model,acupuncture and non-meridian-acupoint groups.Ten 7-week-old male Dahl salt-resistant rats (DR) were used as a blank group.The rats were fed in a SPF animal experiment center.An 8％ NaC1 high salt diet was provided after 1-week adaptive feeding.During the period,rat blood pressure was monitored using an intelligent noninvasive sphygmomanometer.A rat model of prehypertension was successfully made if rat blood pressure rose to 120～ 139/80～89 mmHg after 12 days.At that time,a high salt diet was stopped and a normal diet was provided.The blank and model groups received 15-minute regular fixation and no treatment.The acupuncture group received electroacupuncture at bilateral Quchi and Zusanli.The non-meridianacupoint group received electroacupuncture at fixed control points selected between 10 and 15 mm above bilateral iliac crests 20 mm lateral to the midline.Treatment was given once daily,6 times a week,for five consecutive weeks.The left rat kidney was taken.CaM and eNOS mRNA expressions were determined by quantitative fluorescence PCR.eNOS was quantified and located by western blot and immunohistochemical method.NO content was measured by ELISA.Results As compared with the blank group,CaM and eNOS mRNA expressions,eNOS protein content and NO content decreased significantly in the model group (all P＜0.05).As compared with the model group,CaM and eNOS mRNA expressions,eNOS protein content and NO content increased significantly in the acupuncture group (all P＜0.05).As compared with the model group,CaM expression increased significantly (P＜0.05) but eNOS mRNA expression and NO content had no statistically significant differences (P＞0.05) in the non-meridian-acupoint group.Conclusion The renoprotective effect of electroacupuncture at Quchi and Zusanli may be related to upregulating renal CaM-eNOS-NO signal pathway in DS rats with prehypertension.