NMN在糖尿病肾病大鼠肾脏纤维化中的作用及其机制

doi:10.13481/j.1671-587x.20180608

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (06): 1156-1162.doi: 10.13481/j.1671-587x.20180608

NMN在糖尿病肾病大鼠肾脏纤维化中的作用及其机制

崔奇¹, 王曼伊², 农琳琳³, 翟晓雅¹, 冯乐平¹

1. 桂林医学院公共卫生学院食品卫生与营养教研室, 广西桂林 541004;
2. 桂林医学院第一附属临床医院优生优育科, 广西桂林 541004;
3. 桂林医学院基础医学院实验教学中心, 广西桂林 541004

收稿日期:2018-07-16 出版日期:2018-11-28 发布日期:2018-11-28
通讯作者: 冯乐平,教授,硕士研究生导师(Tel:0773-5835125,E-mail:lpfeng1226@163.com) E-mail:lpfeng1226@163.com
作者简介:崔奇(1994-),女,江苏省盐城市人,在读医学硕士,主要从事糖尿病发病机制的细胞生物学方面的研究。
基金资助:
国家自然科学基金资助课题（81460164，31060161）；广西省科技厅自然科学基金资助课题（2015GXNSF）

Effect of NMN on renal fibrosis of diabetic nephropathy rats and its mechanism

CUI Qi¹, WANG Manyi², NONG Linlin³, ZHAI Xiaoya¹, FENG Leping¹

1. Department of Food Hygiene and Nutrition, School of Public Health, Guilin Medical University, Guilin 541004, China;
2. Department of Eugenic and Superior Nuture, First Affiliated Clinical Hospital, Guilin Medical University, Guilin 541004, China;
3. Experimental Teaching Center, School of Basic Medical Sciences, Guilin Medical University, Guilin 541004, China

Received:2018-07-16 Online:2018-11-28 Published:2018-11-28

摘要/Abstract

摘要： 目的：探讨烟酰胺单核苷酸（NMN）对糖尿病肾病（DN）大鼠肾细胞纤维性变的作用，阐明NMN通过沉默信息调节因子1（Sirt1）和AKT途径缓解肾实质细胞纤维化的机制。方法：采用链脲佐菌素（STZ）制备SD大鼠2型糖尿病模型，模型大鼠随机分为实验组（n=30）和对照组（n=10）：实验组大鼠分为糖尿病+NMN组（n=15，连续皮下注射NMN）和糖尿病+PBS组（n=15，大鼠注射200 μL无菌PBS），之后断头处死大鼠，取肾脏组织进行切片和蛋白提取，Westen blotting法和免疫共聚焦法检测2组大鼠肾小球细胞中Sirt1、AKT、p-FoxO3a和Cav-1蛋白表达水平。采用高浓度葡萄糖（200 mmol·L^-1）处理大鼠肾小球系膜HBZY-1细胞3~6 d后，随机将培养细胞分为4组（分别给予0、50、100和200 μmol·L^-1 NMN），以给予5.6mmol·L^-1葡萄糖不给予NMN的细胞作为对照组，继续培养24 h后收集细胞提取蛋白，采用Westen blotting法检测各组HBZY-1细胞中Sirt1、AKT和p-FoxO3a蛋白表达水平。结果：与糖尿病+PBS组比较，糖尿病+NMN组大鼠肾实质细胞中Sirt1和AKT蛋白表达水平明显升高（P<0.01），糖尿病+NMN组大鼠肾实质细胞中p-FoxO3a和Cav-1蛋白表达水平升高（P<0.01）。与对照组（给予5.6 mmol·L^-1葡萄糖）比较，50 μmol·L^-1NMN组HBZY-1细胞中Sirt1和AKT蛋白表达水平升高（P<0.05），100和200 μmol·L^-1NMN组HBZY-1细胞中Sirt1、AKT和p-FoxO3蛋白表达水平升高（P<0.05或P<0.01）。结论：NMN能够通过调节Sirt1与AKT蛋白表达提高DN大鼠肾小球细胞中内源性p-FoxO3a和Cav-1蛋白表达，NMN及其类似物可能具有潜在预防或治疗DN大鼠肾小球纤维化的作用。

关键词: 糖尿病肾病, 烟酰胺单核苷酸, 沉默信息调节因子1, 蛋白激酶B, 叉头转录因子3a

Abstract: Objective: To investigate the effect of niacinamide mononucleotide (NMN) on the fibrosis of renal cells in the rats with diabetic nephropathy(DN), and to elucidate the mechanism of NMN in regulating the fibrosis of renal parenchymal cells through silent information regulator 1(Sirt1) and AKT pathways.Methods: The rat models of type 2 diabetes mellitus were induced by streptozotocin (STZ) and the model rats were randomly divided into experiment group(n=30) and control group(n=10).The rats in experiment group were divided into diabetes+NMN group(n=15) and diabetes+PBS group(n=15).The rats in diabetes+ NMN group were given subcutaneous injection of NMN for 20 d and the rats in diabetes+PBS group were given 200 μL sterile PBS in the same way.Then the rats were decapitated and the kidney tissues were taken for section and protein extraction. The expression levels of Sirt1, AKT, p-FoxO3a and Cav-1 proteins in kidney tissue of the rats in various groups were detected by Western blotting method and immuno-confocal focusing. The glomerular mesangial HBZY-1 cells were treated with high concentration of glucose (200 mmol·L^-1) for 3-6 d, and then the cells were further randomly divided into 4 groups (treated with 0,50,100,and 200 μmol·L^-1 NMN)and the cells only treated with 5.6 mmol·L^-1 glucose were regareded as control group. After 24 h culture, the cells were collected and the expression levels of Sirt1, AKT, and p-FoxO3a proteins in the HBZY-1 cells in various groups were detected by Western blotting method.Results: Compared with diabetes +PBS group,the expression levels of Sirt1 and AKT proteins in the renal parenchyma cells of the rats in diabetes+ NMN group were significantly increased (P<0.01) and the expression levels of p-FoxO3a and Cav-1 proteins in the renal parenchyma cells of the rats in diabetes + NMN group were also increased(P<0.01). Compared with control group, the expression levels of Sirt1 and AKT proteins in the HBZY-1 cells of the rats in 50 μmol·L^-1 NMN group were significantly increased (P<0.01), and the expression levels of Sirt1, AKT,and p-FoxO3a proteins in the HBZY-1 cells in 100 and 200 μmol·L^-1 NMN groups were increased significantly (P<0.05 or P<0.01).Conclusion: NMN can increase the expression levels of endogenous p-FoxO3a and Cav-1 proteins in the glomerular cells of the DN rats by regulating the expression levels of Sirt1 and AKT proteins, indicating that NMN and its analogues may play an important role in the prevention and treatment of the renal fibrosis of the DN rats.

Key words: diabetic nephropathy, nicotinamide mononucleotide, silent information regulator 1, protein kinase B, Forkhead box O3

中图分类号:

R363.1

崔奇, 王曼伊, 农琳琳, 翟晓雅, 冯乐平. NMN在糖尿病肾病大鼠肾脏纤维化中的作用及其机制[J]. 吉林大学学报(医学版), 2018, 44(06): 1156-1162.

CUI Qi, WANG Manyi, NONG Linlin, ZHAI Xiaoya, FENG Leping. Effect of NMN on renal fibrosis of diabetic nephropathy rats and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2018, 44(06): 1156-1162.

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NMN在糖尿病肾病大鼠肾脏纤维化中的作用及其机制

Effect of NMN on renal fibrosis of diabetic nephropathy rats and its mechanism

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