摘要
目的:基于肾素原受体(PRR)介导的丝裂原活化蛋白激酶(MAPK)信号通路,研究参附强心丸对心肾综合征大鼠心肾凋亡的保护作用机制。方法:Wistar大鼠经肾脏急性缺血再灌注损伤合并腹主动脉缩窄术,制备大鼠心肾综合征(CRS)模型。将CRS大鼠随机分成CRS模型组(ig 10 m L·kg-1纯净水),CRS+参附强心丸组(SFQX组,ig参附强心丸13.2g·kg-1),CRS+柄区肽(HRP)组(iv HRP 10 mg·kg-1),假手术组(ig等体积纯净水)。术后8周给药,1次/d,持续4周。实验结束后,测定血清脑钠肽(BNP),尿素氮(BUN)和肌酐(Cr),小动物超声心动仪测定小动物超声监测舒张末室间隔厚度(IVS),舒张末期左心室后壁厚度(LVPW),左心室射血分数(LVEF),实时荧光PCR测定左心室和肾脏PRR mRNA表达,免疫印迹法(Western blot)测定丝裂原活化蛋白激酶(MAPK)信号通路,包括细胞外信号调控的蛋白激酶(ERK1/2),c-Jun氨基末端激酶(JNK),P38蛋白表达,原位末端标记法(TUNEL)测定左心室和肾脏组织细胞凋亡。结果:与假手术组比较,CRS大鼠血清BNP,BUN,Cr明显升高(P<0.05),IVS,LVPW显著增加(P<0.01),LVEF显著降低(P<0.01),左心室质量指数显著增加(P<0.01),左肾脏质量指数显著减小(P<0.01),组织PRR mRNA高表达(P<0.01),ERK1/2,p38,JNK蛋白表达升高(P<0.01),心肌和肾脏细胞凋亡率达32.5%,63.2%。参附强心丸13.2 g·kg-1可明显减轻CRS大鼠心肌肥厚,抑制IVS,LVPW肥厚,增加EF,血清BUN,Cr明显降低,降低受损组织PRR mRNA表达和ERK1/2,p38蛋白表达,降低心肌和肾脏细胞凋亡率。结论:参附强心丸可增强CRS大鼠心肾功能,通过降低心肾组织PRR mRNA表达,抑制MAPK信号通路ERK1/2,JNK,P38磷酸化,降低心肾细胞凋亡。
Objective： To explore the protective mechanism of Shenfu Qiangxin pill（ SFQX） on heart and kidney cells apoptosis in rats with cardio-renal syndrome（ CRS） based on the（ pro） rennin receptor（ PRR）-mediated mitogen-activated protein kinase（ MAPK） signal pathways. Method： Wistar rats underwent reperfusion injury combined with abdominal aortic constriction after renal ischaemia to prepare CRS models. The CRS rats were divided randomly into three groups： CRS model group（ 10 m L·kg- 1pure water,ig）; CRS ＋ SFQX group（ SFQX group,13. 2 g·kg- 1SFQX,ig）; CRS ＋ Handle region peptide（ HRP） group（ 10 mg·kg- 1HRP,iv）. The rats in sham operation group were ig given with the same volume of pure water. Treatments were given 8 weeks after surgery,1 time / day,for 4 weeks. After the experiment,the rats were detected for brain natriuretic peptide（ BNP）,urea nitrogen（ BUN）,and creatinine（ Cr） levels; ultrasound cardiograph for small animals was used to detect end diastolic interventricular septum thickness（ IVS）,left ventricular end diastolic posterior wall thickness（ LVPW） and left ventricular ejection fraction（ LVEF）; real time fluorescent PCR was used to measure PRR mRNA expressions of left ventricle and kidney; Western blot assay was used to determine MAPK signal pathways,including extracellular signal-regulated kinase 1 /2（ ERK1 /2）,C-Jun amino terminal kinase（ JNK）,and P38 protein expressions; Terminal deoxynucleotidyl transferase d UTP nick and labeling（ TUNEL） was used to detect cells apoptosis of left ventricular and renal tissues. Result： Compared to the sham operation group,serum BNP,BUN and Cr levels were significantly increased（ P〈0. 05）; IVS,LVPW levels were significantly increased（ P〈0. 01）; LVEF levels were significantly decreased（ P〈0. 01）; mass index of left ventricle was significantly increased（ P〈0. 01）; mass index of left kidney was significantly decreased in CRS rats（ P〈0. 01）. Expression of PRR mRNA was increased（ P〈0. 01）; protein expressions of ERK1 /2,p38 and JNK were increased（ P〈0. 01）. Myocardial and renal cell apoptosis rate was 32. 5% and 63. 2% respectively. SFQX 13. 2 g·kg- 1could significantly reduce cardiac hypertrophy in CRS rats,inhibit IVS and LVPW,increase EF,significantly decrease BUN and Cr,decrease PRR mRNA expression and ERK1 /2,P38 protein expression of injured tissues,and decrease the myocardial and renal cell apoptosis rate. Conclusion： SFQX could improve the heart and kidney function of CRS rats,and reduce myocardial and renal cell apoptosis through reducing PRR mRNA expression of myocardial and renal cells and inhibiting ERK1 /2,JNK and P38 phosphorylation in MAPK signal pathways.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2016年第3期121-126,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金青年基金项目(81202801)
