利拉鲁肽通过抑制固醇调节元件结合蛋白-1减轻糖尿病肾病大鼠肾小管异位脂肪沉积  

Liraglutide attenuates renal tubular ectopic fat deposition in diabetic nephropathies rats through inhibiting sterol regulatory element binding protein 1

作　　者：苏可[1] 易波[2] 姚碧晴 夏甜 陈铖[1] Su Ke;Yi Bo;Yao Biqing;Xia Tian;Chen Cheng(Department of Nephrology,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Endocrinology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院肾内科,430060 [2]武汉大学人民医院内分泌科,430060

出　　处：《中华糖尿病杂志》2019年第12期774-781,共8页CHINESE JOURNAL OF DIABETES MELLITUS

基　　金：国家重点研发计划-重大慢病非传染性疾病防控研究专项(2016YFC1305503);国家自然科学基金(81700599);国家科技支撑计划课题(2015BAI12B06);湖北省自然科学基金(2018CFB150)。

摘　　要：目的探讨利拉鲁肽对糖尿病肾病(DN)大鼠异位脂肪沉积的作用及机制。方法40只雄性SD大鼠按照随机数字表法分为三组:正常对照组(10只)、DN溶剂对照组(15只)、DN+利拉鲁肽组治疗组(15只)。利拉鲁肽0.4 mg·kg-1·d-1治疗12周,检测大鼠体重、24 h尿量、24 h尿总蛋白定量、血清胰岛素、血糖、血脂等生化指标,过碘酸雪夫染色光镜观察大鼠肾脏组织病理形态学改变,免疫组化检测大鼠肾组织固醇调节元件结合蛋白-1(SREBP-1)、脂肪分化相关蛋白(ADRP)表达和分布;Western blot检测大鼠肾组织腺苷酸活化蛋白激酶(AMPK)、SREBP-1、ADRP、脂肪酸合成酶(FAS)蛋白表达和p-AMPK磷酸化水平变化。组间比较采用单因素方差分析,两两比较采用最小显著差异法检验。结果DN溶剂对照组大鼠血糖、24 h尿量自造模成功后较糖尿病大鼠造模前显著升高(t=8.630~28.251,均P<0.01)。当利拉鲁肽治疗至12周时DN组大鼠24 h尿总蛋白定量较糖尿病大鼠造模前相比显著升高[(20.3±10.6)比(5.9±2.0)g/24 h,t=3.965,P<0.01];同时出现肾组织系膜增生,甘油三酯(TG)和血清总胆固醇(TC)较正常对照组显著增高(t=3.616、2.363,均P<0.05)。免疫组化结果显示DN溶剂对照组主要表达在肾小管的SREBP-1和ADRP蛋白表达量较正常对照组显著升高(t=7.588、6.160,均P<0.01)。Western blot显示肾组织SREBP-1、FAS、ADRP蛋白表达量较正常对照组显著增加,而AMPK水平下降(t=6.12、8.78、11.16、-12.78,均P<0.01)。糖尿病大鼠经利拉鲁肽治疗12周后,与DN溶剂对照组相比,体重、TG和TC显著下降(t=-3.338、-2.944、-2.390,均P<0.05)、肾组织SREBP-1、FAS、ADRP蛋白表达显著下降,而p-AMPK水平增加(t=-2.833、-4.228、-11.85、7.653,均P<0.05)。结论利拉鲁肽可以抑制DN大鼠肾脏异位脂肪沉积,可能与利拉鲁肽增加AMPK磷酸化活性,抑制SREBP-1转录活性,减少FAS表达,从而抑制脂肪酸合成途径有关。Objective To investigate the effect and mechanism of liraglutide on ectopic fat deposition in rats with diabetic nephropathies(DN).Methods Forty male SD rats were randomly divided into three groups:normal control rats group(n=10),DN vehicle control rats group(n=15)and DN rats with liraglutide group(n=15).DN rats received subcutaneous injection of liraglutide(0.4 mg·kg-1·d-1)for 12 weeks.Body weight,24 h urine volume,24 h total urine protein,serum insulin level,blood glucose,serum triglyceride(TG),total cholesterol(TC),and other biochemical index were detected after 12 week treatment of liraglutide.The histopathological changes of rat kidney were evaluated by periodic acid-schiff staining.The protein expression,distribution of sterol regulatory element binding protein and adipose differentiation-related protein in rat kidney were detected with immunohistochemistry.Renal tissue adenosine monophosphate activated protein kinase(AMPK),sterol regulatory element binding protein 1(SREBP-1),adipose differentiation related protein(ADRP),fatty acid synthase(FAS)and p-AMPK phosphorylation levels protein expression in rats kidney were examined using western blot.One-way ANOVA was used to compare groups,pairwise comparison was tested by the least significant difference method.Results Compared to normal control group,the blood glucose,24 h urine volume,and 24 h total urinary protein levels were increased in the DN vehicle control group(t=8.630-28.251,all P<0.01).When treated with liraglutide for 12 weeks,the total urine protein content of the DN vehicle control rats group was significantly higher than that of the diabetic rats before modeling[(20.3±10.6)vs(5.9±2.0)g/24 h,t=3.965,P<0.01].Glomerular mesangial cell proliferation and matrix expansion,serum TG and TC was significantly increased(t=3.616,2.363,all P<0.05).SREBP-1 and ADRP proteins were mainly distributed in tubular(t=7.588,6.160,all P<0.01),and proteins expression of SREBP-1,FAS and ADRP protein in kidney tissue increased significantly,while AMPK phosphorylation l

关 键 词：糖尿病肾病 脂肪酸合成 固醇调节元件结合蛋白 利拉鲁肽 

分 类 号：R73[医药卫生—肿瘤]