丁苯酞通过下调NF-κB信号通路抑制细胞焦亡减轻大鼠肾缺血-再灌注损伤  

Dl-3-N-butylphthalide alleviates renal ischemia-reperfusion injury by down-regulating NF-κB signaling pathway and inhibiting cell pyroptosis in rat models

作　　者：张瑞波 申开文 王强 袁强 沈俊 Zhang Ruibo;Shen Kaiwen;Wang Qiang;Yuan Qiang;Shen Jun(Department of Urology Surgery,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China)

机构地区：[1]贵州医科大学附属医院泌尿外科,贵阳550004

出　　处：《器官移植》2023年第4期539-546,共8页Organ Transplantation

基　　金：贵州省卫生健康委科学技术基金(gzwkj2021-220);国家自然科学基金培育项目[gyfynsfc(2020)-30];贵州医科大学附属医院2022年博士科研启动基金(gyfybsky-2022-31)。

摘　　要：目的探讨丁苯酞对大鼠肾缺血-再灌注损伤(IRI)的作用机制。方法将40只SD大鼠随机分为假手术组(Sham组)、模型组(IRI组)、NF-κB抑制剂吡咯烷二硫代氨基甲酸酯(PDTC)组、丁苯酞低剂量组(NBP-L组)及丁苯酞高剂量组(NBP-H组),每组8只。检测各组大鼠血清肌酐(Scr)、血清胱抑素C(Cys-C)、血尿素氮(BUN)和血清白细胞介素(IL)-1β、IL-18水平,苏木素-伊红(HE)染色观察各组肾组织病理损伤情况,采用蛋白质印迹法和免疫组织化学法检测肾组织中炎症因子、核因子(NF)-κB信号通路及细胞焦亡相关蛋白表达水平。结果与Sham组比较,IRI组肾组织损伤较为严重,Scr、Cys-C、BUN和血清IL-1β、IL-18水平均升高,蛋白质印迹法结果显示NOD样受体蛋白(NLRP3)、Gasdermin D(GSDMD)、半胱氨酸天冬氨酸蛋白酶(Caspase)-1、IL-18、IL-1β、NF-κB p65、p-NF-κB p65蛋白相对表达量均增加,免疫组织化学染色结果显示NF-κB p65、p-NF-κB p65、IL-1β、IL-18和NLRP3蛋白表达均增多。与IRI组比较,PDTC组、NBP-L组和NBP-H组肾组织的损伤程度均减轻,Scr、Cys-C、BUN和血清IL-18、IL-1β水平均下降,蛋白质印迹法结果显示NLRP3、GSDMD、Caspase-1、IL-1β、IL-18、NF-κB p65、p-NF-κB p65蛋白表达均减少,免疫组织化学染色结果显示NF-κB p65、p-NF-κB p65、IL-1β、IL-18和NLRP3蛋白表达均下降。与NBP-L组比较,NBP-H组肾组织的损伤程度减轻,Scr、Cys-C、BUN和血清IL-18、IL-1β水平均下降,蛋白质印迹法结果显示NLRP3、GSDMD、Caspase-1、IL-1β、IL-18、NF-κB p65、p-NF-κB p65蛋白表达均减少,免疫组织化学染色结果显示NF-κB p65、p-NF-κB p65、IL-1β、IL-18和NLRP3蛋白表达均下降。结论丁苯酞可下调NF-κB/NLRP3信号通路的活性,降低肾IRI后焦亡相关蛋白的表达水平及炎症因子水平,进而抑制细胞焦亡,减轻肾IRI。Objective To elucidate the mechanism of dl-3-N-butylphthalide(NBP)on renal ischemia-reperfusion injury(IRI)in rat models.Methods Forty SD rats were randomly divided into the sham operation group(Sham group),model group(IRI group),NF-κB inhibitor pyrrolidine dithiocarbamate group(PDTC group),low-dose NBP group(NBP-L group)and high-dose NBP group(NBP-H group),with 8 rats in each group.Serum creatinine(Scr),serum cystatin C(Cys-C),blood urea nitrogen(BUN)and serum interleukin(IL)-1βand IL-18 levels were detected in all groups.Pathological injury of renal tissues in each group was observed by Hematoxylin-eosin(HE)staining.The expression levels of inflammatory factors and nuclear factor(NF)-κB signaling pathway and cell pyroptosis-related proteins in renal tissues were measured by Western blot and immunohistochemical staining.Results Compared with the Sham group,renal tissue injury was more severe,and the levels of Scr,Cys-C,BUN and serum IL-1βand IL-18 were all up-regulated in the IRI group.Western blot showed that the relative expression levels of NOD-like receptor protein(NLRP3),Gasdermin D(GSDMD),cysteinyl aspartate specific proteinase(Caspase)-1,IL-18,IL-1β,NF-κB p65 and p-NF-κB p65 proteins were all up-regulated,and immunohistochemical staining revealed that the expression levels of NF-κB p65 and p-NF-κB p65,IL-1β,IL-18 and NLRP3 proteins were all up-regulated in the IRI group.Compared with the IRI group,renal tissue injury was alleviated,and the levels of Scr,Cys-C,BUN and serum IL-18 and IL-1βwere down-regulated in the PDTC,NBP-L and NBP-H groups.Western blot showed that the expression levels of NLRP3,GSDMD,Caspase-1,IL-1β,IL-18,NF-κB p65 and p-NF-κB p65 proteins were down-regulated,and immunohistochemical staining indicated that the expression levels of NF-κB p65,p-NF-κB p65,IL-1β,IL-18 and NLRP3 proteins were down-regulated in the PDTC,NBP-L and NBP-H groups,respectively.Compared with the NBP-L group,renal tissue injury was mitigated,and the levels of Scr,Cys-C,BUN,serum IL-18 and IL-1βwere

关 键 词：丁苯酞 缺血-再灌注损伤 急性肾损伤 细胞焦亡 炎症 NOD样受体蛋白3(NLRP3) 血清肌酐 血尿素氮 核因子(NF)-κB信号通路 血清胱抑素C(Cys-C) 白细胞介素-1β 白细胞介素-18 

分 类 号：R617[医药卫生—外科学] R816.8[医药卫生—临床医学]