葡萄籽多酚对心肌梗死大鼠NF-κB信号通路及肾功能的影响

Effects of Grape Seed Polyphenols on NF-κB Signaling Way and Renal Function in Rats with Myocardial Infarction

目的:观察葡萄籽多酚对心肌梗死大鼠核转录因子κB(NF-κB)信号通路及肾功能的影响。方法:将无特定病原体(SPF)级SD雄雌各半50只大鼠按照体质量随机分为5组,A组(假手术组)、B组(心肌梗死模型组)、C组(心肌梗死模型+低剂量葡萄籽多酚组)、D组(心肌梗死模型+高剂量葡萄籽多酚组)及E组(心肌梗死模型+血府逐瘀胶囊组),C组及D组模型大鼠灌胃由蒸馏水稀释的100 mg/kg及300 mg/kg葡萄籽多酚,E组模型大鼠灌胃250 mg/kg的血府逐瘀胶囊。连续14 d,每日1次。A组及B组大鼠灌胃相同时间、相同次数、相同体积的生理盐水。末次灌胃后,采用超声测定各组大鼠心功能;收集大鼠尿液及血液,采用比色法检测血清肌酐(Cr)、尿素氮(BUN)水平;黄基水杨酸-硫酸钠比浊法检测24 h尿蛋白(UP24);采用苏木精-伊红(HE)染色观察大鼠心肌组织及肾组织病理改变;采用免疫印迹法及实时荧光定量聚合酶链式反应(qRT-PCR)检测肾组织NF-κB、NF-κBp65及IκB激酶β(IκKβ)表达。结果:与A组比较,B组左室舒张末期压力(LVEDP)升高,平均动脉压(MAP)及左室收缩压(LVSP)降低,血Cr、BUN和尿UP24升高(P<0.05);与B组比较,C组、D组及E组大鼠心功能LVEDP降低,MAP及LVSP升高,血Cr、BUN和尿UP24降低(P<0.05)。心肌组织病理:A组大鼠心肌纤维整齐,形态椭圆形,细胞纹理清晰;B组细胞质出现血管水肿出血,心肌组织纤维扩张,出现肌丝溶解;C组、D组及E组细胞质水肿减少,间隙改善,结构接近正常。肾脏组织病理:A组肾脏组织结构完整,肾小管和肾小球结构良好;B组肾小球间质增多,肾间质增宽,炎症浸润严重,肾小管严重萎缩;C组、D组及E组肾组织病理显著改善,肾小球硬度降低,炎症浸润改善。B组肾组织NF-κB、NF-κBp65及IκKβ含量高于A组(P<0.05);C组、D组及E组NF-κB、NF-κBp65及IκKβ含量低于B组(P<0.05)。结论:葡萄籽多酚可改善心肌梗死大鼠心脏功能,提高肾功能,缓解心肌及肾脏组织病理损伤,其机制与抑制NF-κB、NF-κBp65及IκKβ活性相关。

Objective:To explore the effects of grape seed polyphenols on nuclear transcription factorκB(NF-κB)signaling way and renal function in rats with myocardial infarction.Methods:Fifty rats of SD male and female with specific pathogen free(SPF)were randomly divided into five groups according to body weight:group A(sham-operated group),group B(myocardial infarction model group),group C(myocardial infarction model group+low-dose grape seed polyphenol group),group D(myocardial infarction model group+high-dose grape seed polyphenol group)and group E(myocardial infarction model group+Xuefu Zhuyu Capsules group).The rats in the group C and the group D were gavaged with grape seed polyphenols diluted with distilled water at 100 mg/kg and 300 mg/kg for 14 days,once daily,and the rats in group E were gavaged with 250 mg/kg Xuefu Zhuyu Capsules for 14 days,once daily.The rats in the group A and the group B were gavaged with equal volumes of saline at the same time and the same number of times.After final gavage,cardiac function was measured by ultrasound in each group.The serum creatinine(Cr)and blood urea nitrogen(BUN)were detected by colorimetric method in urine and blood.The 24 h urine protein(UP24)was detected by yellow-based salicylic acid-sodium sulfate turbidimetric method.The myocardial,and renal histopathological changes in rats were detected by hematoxylin-eosin(HE)staining.The expressions of NF-κB,NF-κBp65 and IκB kinaseβ(IκKβ)in the renal tissues were detected by immunoblotting and quantitative real-time fluorescence polymerase chain reaction(qRT-PCR).Results:Compared with group A,left ventricular end-diastolic pressure(LVEDP)in the group B was increased,mean arterial pressure(MAP)and left ventricular systolic pressure(LVSP)were decreased,blood Cr,BUN,and urine UP24 were increased(P<0.05).Compared with group B,LVEDP in the group C,group D,and group E was decreased,MAP and LVSP were increased,blood Cr,BUN and urine UP24 were decreased(P<0.05).Myocardial histopathology:group A showed neat myocardial fibers with oval morphology and clear cell texture.Group B showed vascular edema and hemorrhage in cytoplasm showed as follow,dilated myocardial tissue fibers and myofilament lysis,group C,group D,and group E showed decreased cytoplasmic edema,improved gap and near normal structure.Renal histopathology:group A showed intact renal histology,consisting of good tubular and glomerular structures,group B had increased glomerular interstitium,widened interstitium,severe inflammatory infiltration and severe tubular atrophy,and renal histopathology in group C,group D,and group E showed significantly improved.The contents of NF-κB,NF-κBp65 and IκKβin the renal tissues of group B were higher than that in group A(P<0.05),the contents of NF-κB,NF-κBp65 and IκKβin group C,group D,and group E were lower than that in group B(P<0.05).Conclusion:Grape seed polyphenols could improve cardiac function,enhanced renal function and alleviated myocardial and renal histopathological damage in rats with myocardial infarction.Its mechanism was related to inhibition of NF-κB,NF-κBp65 and IκKβactivities.