槲皮素对糖尿病肾病小鼠肾脏P2X7R/NLRP3信号通路和纤维化的影响  

Quercetin Treats Renal Fibrosis in Diabetic Nephropathy via Regulating P2X7R/NLRP3 Pathway in Mice

作　　者：王兴红[1,2] 孙静 马永超[1,2] 孙缦利 李海霞[3] WANG Xinghong;SUN Jing;MA Yongchao;SUN Manli;LI Haixia(Luohe Medical College,Luohe 462000;Henan Special Medical Food Engineering Technology Research Center,Luohe 462000;The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054)

机构地区：[1]漯河医学高等专科学校,漯河462000 [2]河南省特医食品工程技术研究中心,漯河462000 [3]新疆医科大学第一附属医院,新疆乌鲁木齐830054

出　　处：《中药药理与临床》2023年第6期48-53,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：河南省高等学校重点科研项目(编号:20B310007);河南省重点研发与推广专项(科技攻关)项目(编号:232102310501);河南省科技发展计划项目(科技攻关)(编号:1721023610312);漯河医学高等专科学校创新创业发展能力提升工程科技类项目(编号:2021LYZKJXM008)。

摘　　要：目的:观察槲皮素对糖尿病肾病(DN)小鼠肾脏嘌呤能离子通道型受体7(P2X7R)/核苷酸结合寡聚化结构域样受体蛋白3 (NLRP3)信号通路和肾纤维化的影响,揭示槲皮素对DN小鼠肾保护的分子机制。方法:采用腹腔注射链脲佐菌素30 mg/kg建立DN小鼠模型。随机将小鼠分为正常对照组、模型对照组、吡格列酮10 mg/kg组、槲皮素25、50 mg/kg组,每组10只。10 w后测定各组小鼠血清尿素氮(BUN)、肌酐(Scr)、24 h尿蛋白,处死小鼠取双肾计算肾肥大指数;采用酶联免疫吸附法(ELISA)测定血清和尿中白介素-1β(IL-1β)和IL-18的含量;HE染色和PAS染色观察小鼠肾脏病理形态学变化;免疫组化法测定肾组织P2X7R、NLRP3、凋亡相关斑点样蛋白(ASC)和半胱氨酸天冬氨酸酶-1(Caspase-1)蛋白的阳性表达水平;Westernblot检测肾组织α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(CollagenⅠ)和CollagenⅢ的表达。结果:与正常对照组比较,模型对照组小鼠血清BUN、Scr含量、24 h尿蛋白和肾脏肥大指数显著升高(P<0.01);肾小球体积增大,基底膜增厚,系膜基质增多;血清和尿中IL-1β、IL-18含量,肾组织P2X7R、NLRP3、ASC、Caspase-1、α-SMA、CollagenⅠ和CollagenⅢ蛋白表达明显升高(P<0.05或P<0.01);与模型对照组比较,槲皮素25、50 mg/kg组和吡格列酮10 mg/kg组小鼠血清BUN、Scr含量、24 h尿蛋白和肾脏肥大指数显著降低(P<0.01);血清及尿中IL-1β、IL-18含量,肾组织P2X7R、NLRP3、ASC、Caspase-1、α-SMA、CollagenⅠ和CollagenⅢ蛋白表达明显降低(P<0.05或P<0.01),且肾小球病理变化明显改善。结论:槲皮素对DN小鼠肾脏具有保护作用,可能与调控肾脏P2X7R/NLRP3信号通路,抑制炎症反应及肾纤维化发生有关。Objective:To observe the effects of quercetin on the purinergic ligand-gated ion channel 7 receptor(P2X7R)/nucleotide-binding oligomerization domain(NOD)-like receptor family pyrin domain-containing 3(NLRP3) pathway and renal fibrosis in the mouse model of diabetic nephropathy(DN),and reveal the molecular mechanism of the renal protective effect of quercetin on diabetic mice.Methods:The mouse model of DN was established by injection of streptozotocin.Mice were randomized into a normal control group,a model group,a pioglitazone(10 mg/kg) group,and quercetin(25,50 mg/kg) groups,with 10 mice in each group.Ten weeks after administration of corresponding drugs,the blood urea nitrogen(BUN),serum creatinine(Scr),and 24-hour urine protein were measured.After the mice were sacrificed,the kidneys of each mouse were collected for the calculation of the renal hypertrophy index.The levels of interleukin-1β(IL-1β) and IL-18 in the serum and urine were determined by enzyme-linked immunosorbent assay.The pathological changes of the kidney were observed by hematoxylin-eosin staining and periodic acid-Schiff staining.Immunohistochemistry was employed to measure the expression of P2X7R,NLRP3,apoptosis-associated speck-like protein(ASC),and cysteinyl aspartate-specific proteinase-1(Caspase-1) in the renal tissue.Western blotting was employed to determine the protein levels of α-smooth muscle actin(α-SMA) and Collagens Ι and III.Results:Compared with the normal control group,the model group showed increased BUN,Scr,24-hour urine protein,and renal hypertrophy index(P<0.01),enlarged glomeruli,thickened basement membrane,and increased mesangial matrix.Moreover,the modeling elevated the levels of IL-1β and IL-18 in the serum and urine and up-regulated the protein levels of P2X7R,NLRP3,ASC,Caspase-1,α-SMA,and Collagens I and III(P<0.05 or P<0.01).Compared with the model group,quercetin(25,50 mg/kg) and pioglitazone(10 mg/kg) reduced BUN,Scr,24-hour urine protein,and renal hypertrophy index(P<0.01),lowered the levels of IL-1β and I

关 键 词：槲皮素 糖尿病肾病 嘌呤能离子通道型受体7/核苷酸结合寡聚化结构域样受体蛋白3信号通路 炎症 肾纤维化 

分 类 号：R285.5[医药卫生—中药学]