基于OS/NF-κB/炎症因子探讨加味真武汤延缓慢性肾功能衰竭并发心脏病变进展的作用机制  

Based on Oxidative Stress/NF-κB Exploration of Mechanism of Modified Zhenwutang in Delaying Progression of Chronic Renal Failure Complicated with Cardiac Disease by Inflammatory Factors

作　　者：张圆圆 靳培培 宋婧雅 郭登洲[2] ZHANG Yuanyuan;JIN Peipei;SONG Jingya;GUO Dengzhou(Graduate School,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Affiliated Hospital of Hebei University of Chinese Medicine/Hebei Provincial Hospital of Traditional Chinese Medicine,Shijiazhuang 050011,China)

机构地区：[1]河北中医药大学研究生学院,石家庄050091 [2]河北中医药大学第一附属医院/河北省中医院,石家庄050011

出　　处：《中国实验方剂学杂志》2023年第16期100-113,共14页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河北省中医药管理局科研计划项目(2023021);河北省自然科学基金项目(H2022423367);河北中医学院博士研究生创新资助项目(XCXZZBS2023009)。

摘　　要：目的:通过观察加味真武汤对腺嘌呤诱导慢性肾功能衰竭大鼠血清及肾组织中超氧化物歧化酶1(SOD1)、丙二醛(MDA)、晚期氧化蛋白产物(AOPP)、核转录因子-κB p65(NF-κB p65)、磷酸化核转录因子-κB p65(p-p65)、白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α表达及心肾组织病理的影响,探讨加味真武汤延缓慢性肾功能衰竭并发心脏病变进展的可能作用机制。方法:将50只SPF级雄性SD大鼠按照随机数字表法分为正常组10只、造模组40只,将大鼠适应性饲养1周后采用腺嘌呤150 mg·kg^(-1)·d^(-1)灌胃的方法建立实验性慢性肾功能衰竭并发心血管病变大鼠模型。造模完成后随机选取正常组和造模组大鼠各3只取材,检测造模是否成功。造模成功后,将造模组大鼠按照随机数字表法分成模型组、加味真武汤低剂量组、加味真武汤中剂量组、加味真武汤高剂量组、盐酸贝那普利组各6只,进行药物灌胃,每日1次,治疗4周。于实验第17周末检测24 h尿蛋白定量(24 h-UTP)、尿肌酐(UCr),第17周末各组大鼠麻醉后取材,腹主动脉取血后离心取上清检测甘油三酯(TG)、总胆固醇(TC)、血钙、血钾、血磷、肌酐(Cr)、尿素氮(BUN);酶联免疫吸附测定法(ELISA)检测血清AOPP、IL-1β、TNF-α表达水平变化;苏木素-伊红(HE)染色法/马松染色(Masson)法观察心肾组织病理改变;透射电镜观察近端肾小管超微结构变化;免疫组织化学(IHC)法观察肾组织SOD1、MDA、AOPP、NF-κB p65、p-p65、IL-1β、TNF-α表达情况;实时荧光定量聚合酶链式反应法(Real-time PCR)观察肾组织SOD1、NF-κB p65、IL-1β、TNF-αmRNA表达水平;蛋白质免疫印迹法(Western blot)检测肾组织SOD1、MDA、NF-κB p65、p-p65表达水平。结果:(1)与正常组比较,模型组实验大鼠24 h-UTP显著增高(P<0.01),UCr显著降低(P<0.01);模型组实验大鼠Cr、BUN、TG、TC、血磷、血钾显著增高(P<0.01);模型组实验大鼠血清中AOObjective:By observing the effect of modified Zhenwutang on the expression of superoxide dismutase 1(SOD1),malondialdehyde(MDA),advanced oxidation protein product(AOPP),nuclear factor kappa-B(NF-κB)p65,p-p65,IL-1β,TNF-αin serum and renal tissue of adenine-induced chronic renal failure rats and the pathology of heart and kidney tissue,the possible mechanism of modified Zhenwutang delaying the progression of chronic renal failure complicated with heart disease was discussed.Method:Fifty SPF male SD rats were divided into normal group 10 and model group 40 according to the random number table method.After one week of adaptive feeding,the experimental chronic renal failure complicated with cardiovascular disease rat model was established by intragastric administration of adenine 150 mg·kg^(-1)·d^(-1).After the model was completed,3 rats in the normal group and the model group were randomly selected to detect whether the model was successful.After successful modeling,the rats in the model group were divided into model group,modified Zhenwutang low-dose group,modified Zhenwutang medium-dose group,modified Zhenwutang high-dose group and Benazepril hydrochloride group according to the random number table method,with 6 rats in each group.Drugs were administered once a day for 4 weeks.At the end of the 17th week of the experiment,24-hour urinary total protein(24 h-UTP)and urine creatinine(UCr)were detected.At the end of the 17th week,the rats in each group were anesthetized and the abdominal aorta was taken.After centrifugation,the supernatant was taken to detect triglyceride(TG),total cholesterol(TC),serum calcium(Ca),serum potassium,serum phosphate,serum creatinine(Scr),blood urea nitrogen(BUN);the expression levels of serum AOPP,IL-1βand TNF-αwere detected by enzyme linked immunosorbent assay(ELISA).The pathological changes of heart and kidney tissues were observed by hematoxylin-eosin(HE)/Masson method.The ultrastructural changes of proximal renal tubules were observed by transmission electron microscopy.The kid

关 键 词：加味真武汤 腺嘌呤 慢性肾功能衰竭 心脏病变 线粒体损伤 氧化应激 炎症反应 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24