葛根提取物通过调控NOD样受体蛋白3/半胱氨酸天冬氨酸蛋白酶1通路减轻糖尿病大鼠肾损伤的研究

Pueraria lobata extract alleviates kidney injury in diabetic rats by regulating NLRP3/Caspase-1 pathway

目的探讨葛根提取物(EPL)减轻糖尿病大鼠肾损伤的作用机制。方法 60只Wistar大鼠随机分为空白对照(NC)组、模型(Model)组、EPL组、NOD样受体蛋白3(NLRP3)激活剂组(NLRP3)、NLRP3激活剂+EPL(NLRP3+EPL)组,每组各12只。腹腔注射STZ建立糖尿病肾损伤模型大鼠。ELISA法检测血清肌酐(Scr)、BUN、血尿酸(SUA)、24 h UAlb、肾组织丙二醛(MDA)、超氧化物歧化酶(SOD)、白介素1β(IL-1β)和IL-18。HE染色观察肾组织病理学变化。化学荧光法检测肾组织活性氧(ROS)含量。qRT-PCR检测肾组织NLRP3和凋亡相关斑点样蛋白(ASC)mRNA表达。Western blot检测肾组织NLRP3、ASC和半胱氨酸天冬氨酸蛋白酶1(Caspase-1)蛋白表达。结果各组体重和肾组织SOD活力结果显示,Model组低于NC组,EPL组高于Model组,NLRP3+EPL组低于NLRP3组(P<0. 05);FBG、Scr、BUN、SUA、24 h UAlb、MDA、ROS、IL-1β、IL-1、Caspase-1蛋白、NLRP3和ASC蛋白和mRNA水平结果显示,Model组高于NC、EPL组,NLRP3+EPL组高于NLRP3组(P<0. 05)。结论 EPL可减轻糖尿病大鼠肾损伤,可能与抑制NLRP3/Caspase-1通路活性有关。

Objective To explore the possible mechanism of Pueraria lobata extract(EPL)in alleviating renal injury in diabetic rats.Methods group(NC,n=12),model group(Model,n=12),EPL group(n=12),NOD-like receptor protein 3(NLRP3)activator group(NLRP3,n=12)and NLRP3 activator+EPL group(NLRP3+EPL,n=12).Diabetic nephropathy rat model was established by intraperitoneal injection of streptozotocin(STZ). Serum creatinine(Scr),urea nitrogen(BUN),uric acid(SUA),24-hour urinary microprotein content(24 h UAlb)and malondialdehyde(MDA),superoxide dismutase(SOD),interleukin-1β(IL-1β)and interleukin-18(IL-18)were measured by enzyme-linked immunosorbent assay(ELISA). Histopathological changes of kidney were evaluated by Hematoxylin-eosin staining. The content of reactive oxygen species(ROS)in kidney tissues was determined by chemical fluorescence method. The m RNA expression of NLRP3 and ASC in kidney tissue was detected by qRT-PCR. The expression of NLRP3,ASC and Caspase-1 in renal tissue was detected by Western blot.in NC group,higher in EPL group than in Model group,and lower in NLRP3+EPL group than in NLRP3 group(P<0. 05). FBG,Scr,BUN,SUA,24 h UAlb,MDA,ROS,IL-1β,IL-18,Caspase-1 protein,the mRNA and protein of NLRP3 and ASC were higher in Model group than in NC group,lower in EPL group than in Model group,and higher in NLRP3+EPL group than in NLRP3 group(P<0. 05).Conclusion EPL can alleviate renal injury in diabetic rats,and the mechanism may be related to the inhibition of NLRP3/Caspase-1 pathway activity.