摘要
目的:探讨中等强度有氧运动对抑制核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎性小体活化,减轻2型糖尿病小鼠肾脏炎症的作用及其对肾脏的保护作用机制。方法:20只雄性db/db小鼠随机分为糖尿病肾病模型组(DC组,n=10)和有氧运动干预组(DE组,n=10),10只雄性db/m小鼠作为正常对照组(NC组,n=10)。NC组与DC组小鼠正常饲养,DE组小鼠进行跑台运动(0°,5.2米/分钟),60分钟/天,5天/周,共8周。分别于运动干预前、运动干预4周、运动干预8周检测各组小鼠体重、空腹血糖和24小时尿微量白蛋白水平。8周运动干预后,取材测双肾质量、血肌酐和血尿素氮水平;通过肾组织匀浆,测定总超氧化物歧化酶活性和丙二醛水平;利用HE和PAS染色观察肾组织病理变化;采用Western blot法检测肾组织NLRP3、凋亡相关斑点样蛋白(ASC)、含半胱氨酸的天冬氨酸蛋白酶1(Caspase-1)、白介素-1β(IL-1β)、白介素-18(IL-18)、尼克酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)和肾小球裂孔隔膜蛋白分子2(NPHS2)蛋白表达水平。结果:(1)8周正常饲养后,与NC组相比,DC组小鼠体重、空腹血糖、双肾质量、24小时尿微量白蛋白、血肌酐和血尿素氮水平显著上升(P<0.01),肾组织内总超氧化物歧化酶活性明显降低(P<0.01),氧化终产物丙二醛水平显著增加(P<0.01),肾脏病理损伤明显。NLRP3、ASC、Caspase-1、IL-1β、IL-18和NOX4蛋白表达升高(P<0.01),NPHS2蛋白表达下降(P<0.01)。(2)8周运动干预后,与DC组相比,DE组小鼠体重、双肾质量、24小时尿微量白蛋白、血肌酐和血尿素氮指标显著降低(P<0.05,P<0.01),但血糖水平无明显变化。肾组织内总超氧化物歧化酶活性显著升高(P<0.01),氧化终产物丙二醛水平显著下降(P<0.01),肾脏病理损伤减轻。此外,NLRP3、ASC、Caspase-1、IL-1β、IL-18和NOX4蛋白表达显著降低(P<0.01),NPHS2蛋白表达显著升高(P<0.01)。结论:8周中等强度有氧运动可以改善肾功能,减轻肾脏氧化应激水平,抑制NLRP3炎性小体活化,降低炎症反应,减轻2型糖尿病小鼠肾脏损伤。
Objective To determine if moderate-intensity aerobic exercise can prevent nucleotide binding and oligomerization domain-like receptor family pyrin domain-containing protein 3(NLRP3)inflammasome activation and attenuate kidney inflammation,as well as elucidate its mechanism of renal protection in type-2 diabetic mice.Methods Twenty male db/db mice were randomly divided into a diabetic kidney disease model group(DC group,n=10)and an aerobic exercise intervention group(DE group,n=10),while ten db/m mice were selected as a normal control group(NC group,n=10).The mice in the NC and DC groups were fed normally,while those in the DE group performed60-min treadmill exercise(0°,5.2 m/min)every day,5 days a week for 8 weeks.The body weight,fasting blood glucose,and 24-hour urine microalbuminwere measured before as well as after 4-and 8-week intervention.After 8 weeks of exercise intervention,the mice were sacrificed,and the kidney weight,serum creatinine,and blood urea nitrogen levels were measured.The total superoxide dismutase activities and malondialdehyde levels were determined using the renal tissue homogenization.Pathological changes in renal tissue were observed using the Hematoxylin-eosin staining and PAS staining.The protein expression of NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate specific proteinase 1(Caspase-1),interleukin-1β(IL-1β),interleukin-18(IL-18),Nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)and nephrotic syndrome type 2(NPHS2)were detected using the Western blotting.Results(1)Compared with the NC group,the average body weight,fasting blood glucose,kidney weight,urine microalbumin,serum creatinine,and blood urea nitrogen index levels of the DC group increased significantly after 8 weeks of normal feeding(P<0.01).However,the total superoxide dismutase activities in renal tissues decreased significantly,and the malondialdehyde levels increased significantly.The Western blotting results showed that compared with the NC group,the protein expression of NLRP3,ASC,Caspase-1,IL-1β,IL-18,and NOX4 in the DC group increased significantly(P<0.01),but that of NPHS2 decreased significantly(P<0.01).(2)Compared with group DC,the average body weight,kidney weight,urine microalbumin,serum creatinine,and blood urea nitrogen index levels of group DE decreased significantly after 8 weeks of exercise intervention(P<0.05),but the average fasting blood glucose level remained unchanged.Moreover,the total superoxide dismutase activities in renal tissues increased significantly,while the malondialdehyde levels decreased significantly.The average protein expression of NLRP3,ASC,Caspase-1,IL-1β,IL-18,and NOX4 decreased significantly in the DE group(P<0.01),but the NPHS2 protein expression increased significantly(P<0.01).Conclusions Eight weeks of moderate-intensity aerobic exercise can improve the renal function and decrease the renal oxidative stress,NLRP3 inflammasome activation,inflammatory response and renal injury in type-2 diabetic mice.
作者
周中源
史媛媛
王梦
项洁
Zhou Zhongyuan;Shi Yuanyuan;Wang Meng;Xiang Jie(Xuzhou Medical University,Xuzhou 221000,China;Department of Rehabilitation,The Affiliated Hospital of Xuzhou Medical University,Xuzhou 221000,China)
出处
《中国运动医学杂志》
CAS
CSCD
北大核心
2021年第12期962-969,共8页
Chinese Journal of Sports Medicine
基金
国家重点研发计划课题(2020YFC2006604)
江苏省研究生创新工程项目(KYCX21_2674)
关键词
糖尿病肾病
中等强度有氧运动
NLRP3
炎症
氧化应激
diabetic kidney disease
moderate-intensity aerobic exercise
NLRP3
inflammation
oxi-dative stress
作者简介
第一作者:周中源,Email:myszza@163.com;通讯作者:项洁,Email:18052268386@163.com
