野黄芩苷通过CCN1介导抑制核转录因子-κB信号通路改善脂多糖诱导的肾损伤  

Scutellarin alleviates lipopolysaccharide-induced renal injury via mediating cysteine-rich protein 61-connective tissue growth factor-nephroblastoma overexpressed gene 1 expression to inhibit nuclear factor-κB signaling pathway

作　　者：刘雪[1,2] 秦振志 管陈 徐翎钰 代杰 杨成宇 周斌 栾弘 赵龙 张伟[1] 罗从娟[1] 徐岩 Liu Xue;Qin Zhenzhi;Guan Chen;Xu Lingyu;Dai Jie;Yang Chengyu;Zhou Bin;Luan Hong;Zhao Long;Zhang Wei;Luo Congjuan;Xu Yan(Department of Nephrology,the Affiliated Hospital of Qingdao University,Qingdao 266003,Shandong,China;Department of Nephrology,Linyi Central Hospital,Linyi 276400,Shandong,China;Qingdao University,Qingdao 266003,Shandong,China)

机构地区：[1]青岛大学附属医院肾内科,山东青岛266003 [2]临沂市中心医院肾内科,山东临沂276400 [3]青岛大学,山东青岛266003

出　　处：《中华危重病急救医学》2022年第4期400-406,共7页Chinese Critical Care Medicine

基　　金：国家自然科学基金(81970582,81770679);山东省青岛市科技惠民示范引导专项(20-3-4-36-nsh)。

摘　　要：目的探讨野黄芩苷(Scu)对脓毒症相关性急性肾损伤(SA-AKI)的保护作用及其机制。方法①体内实验:将36只雄性C57BL/6小鼠按随机数字表法分为生理盐水(NS)对照组、脂多糖(LPS)致SA-AKI模型组(LPS组)、20 mg/kg Scu对照组(Scu 20对照组)及5、10、20 mg/kg Scu预处理组,每组6只。腹腔注射10 mg/kg LPS制备SA-AKI小鼠模型;NS对照组给予等量NS;Scu预处理组于注射LPS前腹腔注射不同剂量Scu,每日1次、持续1周;Scu 20对照组仅给予20 mg/kg Scu,持续1周。各组于LPS处理24 h后处死小鼠取肾脏,观察肾组织损伤情况;采用蛋白质免疫印迹试验(Western blotting)检测核转录因子-κB(NF-κB)信号通路相关分子、凋亡相关蛋白及富含半胱氨酸蛋白61-结缔组织生长因子-肾母细胞瘤过表达基因1(CCN1)的蛋白表达。②体外实验:体外培养人肾小管上皮细胞株HK-2,待细胞融合至80%时用于实验。在未经LPS处理和经过100 g/L的LPS处理后的细胞中,分别转染pcDNA3.1-CCN1过表达CCN1或小干扰RNA(siRNA)抑制CCN1表达,观察CCN1是否参与了NF-κB信号通路激活和细胞凋亡;同时在转染和未转染CCN1 siRNA的HK-2细胞中加入100 g/L的LPS及20μmol/L的Scu,观察Scu对LPS诱导HK-2细胞损伤保护作用的机制。结果①体内实验结果:LPS诱导SA-AKI小鼠肾功能显著下降,肾小管严重受损;Scu可呈剂量依赖性缓解肾功能及组织学损伤。Western blotting条带分析进一步证实,Scu预处理可呈剂量依赖性降低AKI小鼠肾组织中NF-κB信号通路相关分子及CCN1的蛋白表达,并对细胞凋亡具有显著的缓解作用,说明CCN1参与了NF-κB信号通路激活和细胞凋亡。②体外实验结果:在未经LPS处理的HK-2细胞中,过表达CCN1对凋亡相关蛋白及NF-κB信号通路促炎因子表达无显著影响。而在经过LPS处理的HK-2细胞中,过表达CCN1可显著抑制促炎因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)和单核细胞趋化蛋白-1(MCObjective To explore the protective effect and mechanism of scutellarin(Scu)on sepsis associated-acute kidney injury(SA-AKI).Methods①In vivo experiment:36 male C57BL/6 mice were divided into normal saline(NS)control group,lipopolysaccharide(LPS)induced SA-AKI model group(LPS group),20 mg/kg Scu control group(Scu 20 control group),and 5,10,20 mg/kg Scu pretreatment groups by random number table with 6 mice in each group.The SA-AKI model was reproduced by intraperitoneal injection of 10 mg/kg LPS.The NS control group was injected with NS intraperitoneally.The Scu pretreatment groups were intraperitoneally injected with different doses of Scu every day before LPS injection for 1 week.Scu 20 control group was injected with 20 mg/kg Scu for 1 week.After 24 hours of LPS treatment,mice in each group were sacrificed,kidney tissues were collected,and kidney injury was detected by hematoxylin-eosin(HE)staining.Western blotting was used to detect the protein expression levels of nuclear factor-κB(NF-κB)signaling pathway related molecules,apoptosis-related proteins and cysteine-rich protein 61-connective tissue growth factor-nephroblastoma overexpressed gene 1(CCN1).②In vitro experiment:human renal tubular epithelial cell line HK-2 was cultured in vitro and used for experiment when the cells fused to 80%.In the cells without LPS treatment and after 100 g/L LPS treatment,pcDNA3.1-CCN1 and small interfering RNA(siRNA)CCN1 sequence were transfected to overexpress and inhibit CCN1 expression,respectively,to observe whether CCN1 was involved in NF-κB signaling pathway activation and apoptosis.In addition,100g/L LPS and 20μmol/L Scu were added into HK-2 cells transfected with and without CCN1 siRNA to investigate the mechanism of protective effect of Scu on LPS-induced HK-2 cells injury.Results①The results of in vivo experiment:the renal function of SA-AKI mice induced by LPS was significantly decreased,and had kidney histological damage and severely damaged renal tubules.Scu could alleviate renal function and histologic

关 键 词：野黄芩苷 脓毒症相关性急性肾损伤 CCN1 核转录因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]