鬼箭羽醇提取物通过阻止氧化应激和抑制TNF-α-NF-κB及TβR1-Smad2/3通路减轻兔肾缺血再灌注损伤  

Ethanolic extracts of Euonymus alatus reduce renal ischemia-reperfusion injury in rabbits by preventing oxidative stress and inhibiting TNF-α-NF-κB and TβR1-Smad2/3 signaling pathways

作　　者：陈明环 王咏兰 李相国 吕慧芬 黄德斌 CHEN Ming-huan;WANG Yong-lan;LI Xiang-guo;LÜ Hui-fen;HUANG De-bin(Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu University,Enshi 445000,China)

机构地区：[1]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000

出　　处：《中国病理生理杂志》2022年第4期688-697,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81560675);风湿性疾病发生与干预湖北省重点实验室(湖北民族大学)开放基金项目(No.PT022003)。

摘　　要：目的:探讨鬼箭羽乙醇提取物(EEEA)对家兔肾缺血再灌注损伤(RIRI)的作用及其机制。方法:将30只家兔随机分成正常组、模型组、低剂量EEEA(EEEA-L)组、高剂量EEEA(EEEA-H)组、地塞米松(Dex)组和联合治疗(Dex+EEEA-H)组,每组5只。各组动物分别灌胃药物24 h后,RIRI造模,再继续用相应药物灌胃13 d。其中,正常和模型组家兔灌胃等体积纯水,EEEA-L和EEEA-H组家兔分别灌胃0.156和0.623 g/kg EEEA,Dex组灌胃0.2 mg/kg Dex,Dex+EEEA-H组灌胃0.2 mg/kg Dex+0.623 g/kg EEEA,每天1次。分别于第0、7和14天耳缘静脉采血2 mL用于检测电解质、肾功能[血尿素氮(BUN)和血清肌酐(SCr)]、超氧化物歧化酶(SOD)、丙二醛(MDA)、白细胞介素2/6(IL-2/6)、转化生长因子β_(1)(TGF-β_(1))、肿瘤坏死因子α(TNF-α)等。第14天采血后,以3%戊巴比妥钠(30 mg/kg)耳缘静脉麻醉,取出肾脏,病理学观察及肾组织Paller评分,RT-qPCR和Western blot检测转化生长因子β受体1(TβR1)、Smad2/3和核因子κB(NF-κB)P65的mRNA和蛋白表达。结果:EEEA各组血清K+、Cl−、BUN、SCr、IL-2/6、TGF-β_(1)、TNF-α和MDA水平及肾组织Paller评分均显著低于模型组,而SOD活性高于模型组(P<0.05),且这种作用高剂量显著强于低剂量(P<0.05);病理学也证实EEEA各组较模型组明显好转,较Dex+EEEA-H组肾小管扩张明显;EEEA各组的SOD和MDA水平与Dex+EEEA-H无显著差异(P>0.05);EEEA各组和Dex+EEEA-H组TβR1和NF-κB P65的mRNA和蛋白表达显著低于模型组(P<0.05),EEEA-H组与Dex+EEEA-H组无显著差异(P>0.05)。结论:EEEA对RIRI家兔的肾功能有保护作用,其机制可能与其多成分多靶位互补作用,阻止了氧化应激反应并抑制了TNF-α-NF-κB和TβR1-Smad2/3信号通路有关。AIM:To explore the effects of ethanolic extracts of Euonymus alatus(EEEA)on renal ischemia-reperfusion injury(RIRI)in rabbits and the mechanism.METHODS:Thirty rabbits were randomly divided into 5 groups(n=5),including normal group,model group,low-dose EEEA(EEEA-L)group,high-dose EEEA(EEEA-H)group,dexamethasone(Dex)group,and combination therapy(Dex+EEEA-H)group.All animals were given corresponding drugs by intragastric gavage for 24 h before RIRI modeling,and then they were continuously gavaged by the corresponding drugs for 13 d,once a day.The rabbits in normal and model groups were given equal volume of pure water,while those in EEEA-L and EEEA-H groups were given 0.156 and 0.623 g/kg EEEA,respectively.The rabbits in Dex group were given 0.2 mg/kg Dex,while those in Dex+EEEA-H group were given 0.2 mg/kg Dex and 0.623 g/kg EEEA.On days 0,7 and 14,2 mL blood was collected from the ear veins of the rabbits for separating serum to detect electrolytes,superoxide dismutase(SOD),malondialdehyde(MDA),interleukin-2/6(IL-2/6),transforming growth factor-β_(1)(TGF-β_(1)),tumor necrosis factor-α(TNF-α)and renal function[blood urea nitrogen(BUN)and serum creatinine(SCr)].After blood collection on the 14th day,the rabbits were anesthetized by injecting from the ear veins with 3%pentobarbital sodium at 30 mg/kg,and the kidneys were removed for pathological observation with renal tissue Paller scoring,and detection of transforming growth factor-βreceptor 1(TβR1),Smad2/3 and nuclear factor-κB(NF-κB)P65 mRNA and protein expression by RT-qPCR and Western blot.RESULTS:Serum K^(+),Cl^(−),BUN,SCr,IL-2/6,TGF-β_(1),TNF-αand MDA levels,and renal tissue Paller scores in EEEA groups were lower than those in model group,while SOD activity was higher than that in model group(P<0.05).These effects were significantly stronger at high dose than low dose(P<0.05).Pathological observation confirmed that the renal tissue in EEEA groups was significantly better than that in model group,and renal tubule dilatation was more obvious than th

关 键 词：鬼箭羽乙醇提取物 肾缺血再灌注损伤 核因子ΚB 肿瘤坏死因子α 转化生长因子β受体1 Smad2/3蛋白 

分 类 号：R285.5[医药卫生—中药学] R692[医药卫生—中医学]