| 熊 锐,熊哲学,唐明娟,韦宜东,李凝旭.达格列净通过增强营养剥夺信号促进糖尿病肾病大鼠足细胞自噬的研究[J].,2023,(12):2201-2208 |
| 达格列净通过增强营养剥夺信号促进糖尿病肾病大鼠足细胞自噬的研究 |
| The Study of Dapagliflozin Promotes Podocyte Autophagy in Diabetic Kidney Disease Rats by Enhancing Nutrient Deprivation Signaling |
| 投稿时间：2022-12-23 修订日期：2023-01-18 |
| DOI：10.13241/j.cnki.pmb.2023.12.001 |
| 中文关键词: 糖尿病肾病 达格列净 营养剥夺信号 自噬 |
| 英文关键词: Diabetic kidney disease Dapagliflozin Nutrient deprivation signaling Autophagy |
| 基金项目:国家重点研发计划基金资助项目（SQ2018YFC200194-02） |
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| 中文摘要: |
| 摘要 目的：探究达格列净是否通过增强营养剥夺信号促进糖尿病大鼠肾脏足细胞自噬，延缓糖尿病肾病进展。方法：采用高脂饮食和低剂量链脲佐菌素（STZ）腹腔注射诱导糖尿病模型。将成模大鼠随机分为糖尿病组（n=6），达格列净组（n=7），另设正常对照组（n=7）。达格列净组和糖尿病组分别给予达格列净和生理盐水灌胃4周。灌胃结束后收集大鼠24 h尿液以及血液、肾脏。记录体重、肾重、肾周脂肪重并测定空腹血糖、空腹血清胰岛素、血肌酐、尿素氮、甘油三酯以及尿白蛋白、尿总蛋白、尿白蛋白/肌酐比值；ELISA法检测血清酮体水平；HE染色、PAS染色以及电镜观察肾脏组织病理学改变；免疫组化和免疫荧光检测各组自噬相关蛋白7（ATG7）和足细胞标记蛋白NPHS1的表达情况； Western blot检测各组肾皮质中ATG7、沉默信息调节因子同源蛋白1（SIRT1）、过氧化物酶体增殖物激活受体γ共激活物-1α（PGC-1α）、成纤维细胞生长因子21（FGF21）、磷酸烯醇丙酮酸羧激酶（PEPCK）、过氧化物酶体增殖物激活受体α（PPARα）的蛋白水平。结果：与糖尿病组相比，达格列净组空腹血糖、24 h尿白蛋白、24 h尿总蛋白、尿白蛋白/肌酐比值降低，肾重/体重比、肾周脂肪重/体重比减小（P<0.05）；HE和PAS染色和电镜观察到糖尿病组较达格列净组肾小球基底膜增厚，足细胞足突增宽、融合（P<0.05）；达格列净组甘油三酯、空腹血清胰岛素以及胰岛素抵抗指数低于糖尿病组，血清酮体高于糖尿病组（P<0.05）；电镜下观察到达格列净组足细胞内自噬溶酶体数密度高于糖尿病组（P<0.05）；免疫组化和免疫荧光显示达格列净组足细胞NPHS1和ATG7的表达高于糖尿病组（P<0.05）； Western blot显示达格列净组肾皮质ATG7、SIRT1/PGC-1α/FGF21信号通路以及PEPCK、PPARα的表达较糖尿病组明显升高（P<0.05）。结论：达格列净增强营养剥夺信号的同时，观察到糖尿病大鼠肾脏足细胞自噬增强，这种自噬的增强可能是通过营养剥夺信号诱导的，其中的机制可能与达格列净上调SIRT1/PGC-1α /FGF21信号通路有关。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate whether dapagliflozin can enhance nutrient deprivation signaling and promote autophagy of kidney cells in diabetic rats, and delay the progression of diabetic nephropathy. Methods: Diabetes model was induced by high-fat diet and low-dose streptozotocin (STZ) intraperitoneal injection. The model rats were randomly divided into diabetes control group (n=6), dapagliflozin group (n=7), and normal control group (n=7). Dapagliflozin group was treated with dapagliflozin by gavage for 4 weeks, while diabetes control group was treated with normal saline by gavage for 4 weeks. At the end of gavage, blood, kidney and the urine of 24 hours were collected. Body weight, kidney weight and perirenal fat weight were recorded, and fasting blood glucose, serum insulin, serum creatinine, urea nitrogen, triglyceride, urinary albumin, total urinary protein and urinary albumin/creatinine ratio were measured. Serum ketone body levels were detected by ELISA. HE staining, PAS staining and electron microscopy were used to observe the pathological changes of kidney. The expressions of autophagy related protein 7 (ATG7) and podocyte marker NPHS1 in each group were detected by immunohistochemistry and immunofluorescence. Western blot was used to detect the protein levels of ATG7, silent mating type information regulation 2 homolog 1 (SIRT1), peroxisome proliferators-activated receptor γ coactivator-1α(PGC-1α), fibroblast growth factor21 (FGF21), phosphoenolpyruvate carboxykinasen (PEPCK) peroxisome proliferators-activated receptor (PPARα) in renal cortex of each group. Results: Compared with diabetes control group, fasting blood glucose, 24 h urinary albumin, 24 h urinary total protein, urinary albumin/creatinine ratio, renal weight/body weight ratio and perirenal fat weight/body weight ratio were decreased in dapagliflozin group (P<0.05). HE staining, PAS staining and electron microscope showed that compared with dapagliflozin group, the glomerular basement membrane was thickened, the podocyte process was widened and fused in diabetes control group (P<0.05). Triglyceride, fasting insulin, homeostasis model assessment-insulin resistance index in dapagliflozin group were lower than those in diabetes control group, and serum ketone body was higher than that in diabetes control group (P<0.05). Electron microscope showed that the numerical density of autolysosomes in dapagliflozin group was higher than that in diabetes control group. Immunohistochemistry and immunofluorescence showed that the expressions of NPHS1 and ATG7 in podocyte of dapagliflozin group were higher than those in diabetes control group. Western blot showed that the expression of ATG7, SIRT1/PGC-1α/FGF21 signaling pathway and PEPCK, PPARα in the renal cortex of dapagliflozin group were significantly higher than those of diabetes control group. Conclusion: Dapagliflozin enhances nutrient deprivation signaling and promote renal podocyte autophagy in diabetic rats, the enhancement of autophagy may be induced by nutrient deprivation signaling, and the mechanism may be related to the up-regulation of SIRT1/PGC-1 α/FGF21 signaling pathway. |
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