常春藤素皂苷元介导Axin2/AREG轴抑制炎症缓解小鼠急性肾损伤  

Hederagenin mediates Axin2/AREG axis to inhibit inflammation and alleviate acute kidney injury in mice

作　　者：徐玲慧 梁颖兰 粟宏伟 李健春 李贵平 王丽 邹远霞 XU Linghui;LIANG Yinglan;SU Hongwe;LI Jianchun;Ll Guiping;WANGLi;ZOU Yuanxia(Research Center of Integraterl Chinese anrd Western Melicine,the Affiliated Traditional Chinese.MedicineHospital,SouthwestMedlical Univensity,Lnuzhou 646000,China;Department of Nephrology,the Affiliated Trarditional Chinese Melicine Hospital,Southwest Medlical University,Lnzhou 646000,China;Department ofUmology,the Affiliated Tralitional Chinese Medlicine Hospital,Southwest Medlical University,Luzhou 646000,China;Children's Diagnosis and Treatiment Center,the Affiliated Tnalitional Chinese Merlicine Hospital,Southwest Medlical University,Luzhou 646000,China)

机构地区：[1]西南医科大学附属中医医院中西医结合研究中心,四川泸州646000 [2]西南医科大学附属中医医院肾病科,四川泸州646000 [3]西南医科大学附属中医医院泌尿外科,四川泸州646000 [4]西南医科大学附属中医医院儿童诊疗中心,四川泸州646000

出　　处：《中国实验动物学报》2025年第2期157-168,共12页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家自然科学基金(82104665);四川省科技计划资助(2022YFS0621);西南医科大学科技项目资助(2023ZYYQ01)。

摘　　要：目的 探讨常春藤素皂苷元(hederagenin, HDG)对顺铂(cisplatin, Cis)诱导的急性肾损伤(acute kidney injury, AKI)小鼠的保护作用及潜在机制。方法 将24只雄性C57BL/6J小鼠随机分为对照组、AKI模型组、HDG低剂量组HDG高剂量组,每组各6只。通过腹腔注射Cis 20 mg/kg建立小鼠AKI模型,HDG低剂量组及HDG高剂量组分别给予20、40 mg/kg灌胃HDG,3 d后进行取材。收集小鼠肾进行苏木素-伊红(HE)及糖原(periodic acid-schiff, PAS)染色评估肾病理情况,收集血清检测肌酐、尿素氮的变化,Western Blot检测p-P65、P65、IL-6、TNF-α、IL-1β等炎症相关蛋白的表达。体外使用Cis 200 ng/mL刺激肾小管上皮细胞(TCMK1)建立炎症细胞模型,设置空白组、Cis模型组、HDG低剂量组、HDG高剂量组、Axin2过表达组、HDG+Axin2过表达组,收取细胞蛋白后检测p-P65、P65、IL-6、TNF-α、IL-1β、Axin2、AREG的表达变化。结果与对照组相比,AKI模型组小鼠血清肌酐和尿素氮水平明显升高(P<0.05),并伴随肾小管空泡变性、炎细胞浸润及糖原沉积等病理改变,炎症相关蛋白p-P65、TNF-α、IL-6、IL-1β及Axin2表达均显著上调(P<0.05)。HDG干预后,血清肌酐和尿素氮水平呈剂量依赖性下降(高剂量组降幅>低剂量组降幅,P<0.05),肾组织病理损伤明显减轻,上述炎症因子及Axin2表达同步降低(P<0.05)。体外实验证实HDG可剂量依赖性抑制Gis诱导的TCMK1细胞中Axin2及相关炎症因子表达。转录组测序显示Axin2过表达显著上调双调蛋白(AREG)表达(P<0.05)。机制研究表明,HDG通过抑制Axin2/AREG轴降低p-P65磷酸化水平(P<0.05),而Axin2过表达可逆转HDG对Cis诱导肾小管细胞损伤的保护作用。结论 HDG通过抑制Axin2/AREG轴的激活减轻炎症,缓解AKI小鼠肾损伤。Objective To investigate the protective effect of hederagenin(HDG)on cisplatin(Cis)⁃induced acute kidney injury(AKI)in mice and its potential mechanism.Methods 24 male C57BL/6J mice were randomly divided into a control group,AKI model group,HDG low⁃dose group,and HDG high⁃dose group,with six mice in each group.AKI model was established by intraperitoneal injection of 20 mg/kg cisplatin(Cis).The HDG low⁃dose and HDG high⁃dose groups were given 20,40 mg/kg HDG by intragastric administration,respectively,and samples were collected 3 days later.The kidneys of the mice were collected for hematoxylin⁃eosin(HE)and periodic⁃acid⁃schiff(PAS)staining to evaluate the kidney pathology,and serum was collected to detect changes in serum creatinine(Scr)and blood urea nitrogen(BUN).The expression of p⁃P65,P65,IL⁃6,TNF⁃α,IL⁃1β,and other inflammatory⁃related proteins was detected by Western Blot.A TCMK1(renal tubular epithelial cell)inflammatory cell model was established by Cis(200 ng/mL)stimulation in vitro.Blank group,Cis model group,HDG low⁃dose group,HDG high⁃dose group,Axin2 overexpression group,HDG+Axin2 overexpression group were set up.In the Axin2⁃overexpression group,the expression of p⁃P65,P65,IL⁃6,TNF⁃α,IL⁃1β,Axin2,and AREG was detected among total cell proteins.Results Compared with the control group,AKI model mice exhibited significantly elevated serum creatinine and blood urea nitrogen levels(P<005),accompanied by pathological alterations including vacuolar degeneration of renal tubules,inflammatory cell infiltration,and glycogen deposition,and the expression of inflammation⁃related proteins(p⁃P65,TNF⁃α,IL⁃6,IL⁃1β)and Axin2 was markedly upregulated in AKI mice(P<005).HDG treatment induced a dose⁃dependent reduction in serum creatinine and blood urea nitrogen levels(high⁃dose>low⁃dose,P<005),alleviated renal histopathological damage,and concurrently suppressed the expression of these inflammatory mediators and Axin2(P<005).HDG was confirmed that dose⁃dep

关 键 词：常春藤素皂苷元 急性肾损伤 Axin2 AREG 炎症 

分 类 号：Q95-33[生物学—动物学]