利拉鲁肽对糖尿病肾病大鼠的肾保护作用及NLRP3炎症小体的影响
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摘要
目的探讨利拉鲁肽对糖尿病肾病(DN)大鼠肾脏的保护作用及对NOD样受体3(NLRP3)炎症小体的影响。方法采用高糖高脂饮食联合小剂量链脲佐菌素建立大鼠DN模型,26只DN大鼠随机分为模型组和观察组,每组13只,同时取正常SD大鼠10只设为正常对照组。观察组予以利拉鲁肽400μg·kg-1皮下注射,每日1次,模型组和正常对照组给予等容量生理盐水皮下注射,连续给药8周。检测各组空腹血糖(FPG)、血尿素氮(BUN)、血肌酐(Scr)、血清白细胞介素1β(IL-1β)和白细胞介素18(IL-18)的水平以及24 h尿蛋白定量(UTP)。比较各组肾组织中NLRP3、凋亡相关斑点样蛋白(ASC)和天冬氨酸特异性半胱氨酸蛋白酶1(caspase-1)的蛋白和m RNA表达水平。结果模型组FPG、Scr、BUN、IL-1β、IL-18和UTP水平均高于正常对照组(P<0.05),观察组FPG、Scr、BUN、IL-1β、IL-18和UTP水平均低于模型组(P<0.05)。模型组肾组织中NLRP3、ASC、caspase-1蛋白和m RNA表达水平均高于正常对照组(P<0.05),观察组肾组织中NLRP3、ASC、caspase-1蛋白和m RNA表达水平均低于模型组(P<0.05)。结论利拉鲁肽对DN大鼠具有肾保护作用,该作用可能通过下调NLRP3炎症小体的表达而实现。
AIM To investigate renal protection of liraglutide on diabetic nephropathy(DN) rats and its effect on Nod-like receptor pyrin domain 3(NLRP3) inflammasome.METHODS DN model was induced by high glucose and high fat diet combined with small dose streptozotocin.Twenty-six DN rats were divided into model group and observation group by using random digital table,with 13 rats in each group.And 10 normal rats were selected as normal control group.The observation group was given liraglutide 400 μg·kg-1 subcutaneous injection,once a day.The model group and normal control group were given normal saline subcutaneous injection.The drug was given for 8 weeks.The fasting plasma glucose(FPG),serum creatinine(Scr),blood urea nitrogen(BUN) and 24 h urinary total protein(UTP) were detected by automatic biochemistry analyzer.The expression of interleukin-1 beta(IL-1β) and interleukin-18(IL-18) in serum was detected by ELISA.The expression of NLRP3,apoptosis-associated speck-like protein containing CARD(ASC),caspase-1 protein and m RNA were detected by Western blot and reverse transcriptase polymerase chain reaction(RT-PCR).RESULTS The levels of FPG,Scr,BUN,IL-1β,IL-18 and UTP in the model group were significantly higher than those in the normal control group(P < 0.05).The levels of FPG,Scr,BUN,IL-1β,IL-18 and UTP in the observation group were significantly lower than those in the model group(P < 0.05).The levels of NLRP3,ASC,caspase-1 protein and m RNA in the renal tissue of the model group were higher than those in the normal control group with statistical difference(P < 0.05).The levels of NLRP3,ASC,caspase-1 protein and m RNA in the renal tissue of the observation group were lower than those in the model group with statistical difference(P < 0.05).CONCLUSION Liraglutide has renal protective effect on DN rats,and the effect may be achieved through down-regulation of NLRP3 inflammasome.
AIM To investigate renal protection of liraglutide on diabetic nephropathy(DN) rats and its effect on Nod-like receptor pyrin domain 3(NLRP3) inflammasome.METHODS DN model was induced by high glucose and high fat diet combined with small dose streptozotocin.Twenty-six DN rats were divided into model group and observation group by using random digital table,with 13 rats in each group.And 10 normal rats were selected as normal control group.The observation group was given liraglutide 400 μg·kg-1 subcutaneous injection,once a day.The model group and normal control group were given normal saline subcutaneous injection.The drug was given for 8 weeks.The fasting plasma glucose(FPG),serum creatinine(Scr),blood urea nitrogen(BUN) and 24 h urinary total protein(UTP) were detected by automatic biochemistry analyzer.The expression of interleukin-1 beta(IL-1β) and interleukin-18(IL-18) in serum was detected by ELISA.The expression of NLRP3,apoptosis-associated speck-like protein containing CARD(ASC),caspase-1 protein and m RNA were detected by Western blot and reverse transcriptase polymerase chain reaction(RT-PCR).RESULTS The levels of FPG,Scr,BUN,IL-1β,IL-18 and UTP in the model group were significantly higher than those in the normal control group(P < 0.05).The levels of FPG,Scr,BUN,IL-1β,IL-18 and UTP in the observation group were significantly lower than those in the model group(P < 0.05).The levels of NLRP3,ASC,caspase-1 protein and m RNA in the renal tissue of the model group were higher than those in the normal control group with statistical difference(P < 0.05).The levels of NLRP3,ASC,caspase-1 protein and m RNA in the renal tissue of the observation group were lower than those in the model group with statistical difference(P < 0.05).CONCLUSION Liraglutide has renal protective effect on DN rats,and the effect may be achieved through down-regulation of NLRP3 inflammasome.
引文
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[9]夏小慧,李伟,周冬梅,等.利拉鲁肽对糖尿病大鼠肾组织的保护作用及相关机制[J].中华肾脏病杂志,2016,32(3):200-205.
[10]杨烨,张园园,田燕燕,等.活性维生素D3对糖尿病肾病大鼠转化生长因子β1及相关因子的调节作用[J].中华内分泌代谢杂志,2015,31(1):66-70.
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[12]ZHANG MH,FENG L,ZHU MM,et al.The anti-inflammation effect of Moutan Cortex on advanced glycation end productsinduced rat mesangial cells dysfunction and high-glucose-fat diet and streptozotocin-induced diabetic nephropathy rats[J].J Ethnopharmacol,2014,151(1):591-600.
[13]WEN H,MIAO EA,TING JP.New mechanisms of NOD-like receptor-associated inflammasome activation[J].Immunity,2013,39(3):432-441.
[14]von SCHOLTEN BJ,HANSEN TW,GOETZE JP,et al.Glucgonlike peptide 1 receptor agonist(GLP-1 RA):long-term effect on kidney function in patients with 2 diabetes[J].J Diabetes Complications,2015,29(5):670-674.
[15]刘静芹,何玉秀,张桂玲,等.利拉鲁肽对糖尿病的心肌保护效应及机制研究进展[J].中国新药与临床杂志,2016,35(9):625-630.
[16]温智峰,蒲蔚荣,刘香春,等.利拉鲁肽对不同海拔地区糖调节受损肥胖患者血管内皮功能障碍的影响[J].中国新药与临床杂志,2016,35(8):576-580.
[17]李里,卢松.利拉鲁肽对糖尿病大鼠肾脏病变的影响[J].药物研究,2014,23(14):20-22.
[18]WANG C,PAN Y,ZHANG QY,et al.Quercetin and allopurinol ameliorate kidney injury in STZ-treated rats with regulation of renal NLRP3 inflammasome activation and lipid accumulation[J].PLo S One,2012,7(6):e38285.
[19]SHAHZAD K,BOCK F,DONG W,et al.Nlrp3-inflammasome activation in non-myeloid-derived cells aggravates diabetic nephropathy[J].Kidney Int,2015,87(1):74-84.
[20]ZHANG C,BOINI KM,XIA M,et al.Activation of Nod-like receptor protein 3 inflammasomes turns on podocyte injury and glomerular sclerosis in hyperhomocysteinemia[J].Hypertension,2012,60(1):154-162.
[21]王海芳,曹情雯,牛银波,等.利拉鲁肽通过调控HSP70-TLR4轴抑制db/db小鼠肾脏的炎症性损伤[J].免疫学杂志,2017,33(3):224-231.
[22]于莉莉,张国俊,宋向凤.经典炎症小体的激活机制与功能[J].中华微生物学和免疫学杂志,2015,35(1):73-78.
[23]张曼曼,张蓓蓓,王林,等.Toll样受体4及相关炎性信号分子在高脂饮食诱导的肥胖小鼠肾脏组织水平升高[J].细胞与分子免疫学杂志,2015,31(9):1170-1174.
[24]丁丽红.蛋白尿引起肾小管间质损伤新机制——NLRP3炎性体及TLR2、TLR4信号通路激活[D].南京:东南大学,2015.
[25]江思瑜,梁婵,吴晓光,等.利拉鲁肽对糖尿病大鼠肾损伤过程中My D88蛋白表达及炎症因子的影响[J].中国药理学与毒理学杂志,2016,30(10):1052.
[2]中华医学会糖尿病学分会微血管并发症学组.糖尿病肾病防治专家共识(2014年版)[J].中国糖尿病杂志,2014,6(11):792-801.
[3]WHITING D,GUARIGUATA L,WEIL C,et al.IDF daibetes atlas:global estimates of the prevalence of diabetes for 2011 and2030[J].Diabetes Res Clin Pract,2011,94(3):311-321.
[4]许山荣,钟一红,陈波,等.上海市郊区2型糖尿病患者肾脏疾病及其危险因素研究[J].中华内科杂志,2012,51(1):18-23.
[5]ROBBINS GR,WEN H,TING JP.Inflammasomes and metabolic disorders:old genes in modern diseases[J].Mol Cell,2014,54(2):297-308.
[6]HANEKLAUS M,O'NEILL LA.NLRP3 at the interface of metabolism and inflammation[J].Immunol Rev,2015,265(1):53-62.
[7]MAN SM,KANNEGANTI TD.Gasdermin D:the long-awaited executioner of pyroptosis[J].Cell Res,2015,25(11):1183-1184.
[8]王定坤,陈广,陆付耳.NLRP3炎症小体与2型糖尿病的研究进展[J].生理科学进展,2014,45(2):111-114.
[9]夏小慧,李伟,周冬梅,等.利拉鲁肽对糖尿病大鼠肾组织的保护作用及相关机制[J].中华肾脏病杂志,2016,32(3):200-205.
[10]杨烨,张园园,田燕燕,等.活性维生素D3对糖尿病肾病大鼠转化生长因子β1及相关因子的调节作用[J].中华内分泌代谢杂志,2015,31(1):66-70.
[11]HARJUTSALO V,GROOP PH.Epidemiology and risk factors for diabetic kidney disease[J].Adv Chronic Kidney Dis,2014,21(3):260-266.
[12]ZHANG MH,FENG L,ZHU MM,et al.The anti-inflammation effect of Moutan Cortex on advanced glycation end productsinduced rat mesangial cells dysfunction and high-glucose-fat diet and streptozotocin-induced diabetic nephropathy rats[J].J Ethnopharmacol,2014,151(1):591-600.
[13]WEN H,MIAO EA,TING JP.New mechanisms of NOD-like receptor-associated inflammasome activation[J].Immunity,2013,39(3):432-441.
[14]von SCHOLTEN BJ,HANSEN TW,GOETZE JP,et al.Glucgonlike peptide 1 receptor agonist(GLP-1 RA):long-term effect on kidney function in patients with 2 diabetes[J].J Diabetes Complications,2015,29(5):670-674.
[15]刘静芹,何玉秀,张桂玲,等.利拉鲁肽对糖尿病的心肌保护效应及机制研究进展[J].中国新药与临床杂志,2016,35(9):625-630.
[16]温智峰,蒲蔚荣,刘香春,等.利拉鲁肽对不同海拔地区糖调节受损肥胖患者血管内皮功能障碍的影响[J].中国新药与临床杂志,2016,35(8):576-580.
[17]李里,卢松.利拉鲁肽对糖尿病大鼠肾脏病变的影响[J].药物研究,2014,23(14):20-22.
[18]WANG C,PAN Y,ZHANG QY,et al.Quercetin and allopurinol ameliorate kidney injury in STZ-treated rats with regulation of renal NLRP3 inflammasome activation and lipid accumulation[J].PLo S One,2012,7(6):e38285.
[19]SHAHZAD K,BOCK F,DONG W,et al.Nlrp3-inflammasome activation in non-myeloid-derived cells aggravates diabetic nephropathy[J].Kidney Int,2015,87(1):74-84.
[20]ZHANG C,BOINI KM,XIA M,et al.Activation of Nod-like receptor protein 3 inflammasomes turns on podocyte injury and glomerular sclerosis in hyperhomocysteinemia[J].Hypertension,2012,60(1):154-162.
[21]王海芳,曹情雯,牛银波,等.利拉鲁肽通过调控HSP70-TLR4轴抑制db/db小鼠肾脏的炎症性损伤[J].免疫学杂志,2017,33(3):224-231.
[22]于莉莉,张国俊,宋向凤.经典炎症小体的激活机制与功能[J].中华微生物学和免疫学杂志,2015,35(1):73-78.
[23]张曼曼,张蓓蓓,王林,等.Toll样受体4及相关炎性信号分子在高脂饮食诱导的肥胖小鼠肾脏组织水平升高[J].细胞与分子免疫学杂志,2015,31(9):1170-1174.
[24]丁丽红.蛋白尿引起肾小管间质损伤新机制——NLRP3炎性体及TLR2、TLR4信号通路激活[D].南京:东南大学,2015.
[25]江思瑜,梁婵,吴晓光,等.利拉鲁肽对糖尿病大鼠肾损伤过程中My D88蛋白表达及炎症因子的影响[J].中国药理学与毒理学杂志,2016,30(10):1052.