| 温玉,熊伟,刘城,等.基于 AMPK/SIRT1-FoxO1通路探讨蚕沙提取物 DNJ对糖尿病小鼠肾纤维化和糖脂代谢的影响[J].安徽医药,2025,29(6):1114-1120. |
| 基于 AMPK/SIRT1-FoxO1通路探讨蚕沙提取物 DNJ对糖尿病小鼠肾纤维化和糖脂代谢的影响 |
| Investigating the effect of DNJ extracted from silkworm on renal fibrosis and glycolipid metabolism in diabetic mice via the AMPK/SIRT1-FoxO1 pathway |
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| DOI：10.3969/j.issn.1009-6469.2025.06.010 |
| 中文关键词: 蚕沙 糖尿病肾病 1-脱氧诺吉霉素 AMP活化的蛋白质激酶( AMPK)/沉默信息调节因子 1(SIRT1)-叉头框蛋白 O1(FoxO1)通路 肾纤维化 糖脂代谢 |
| 英文关键词: Silk-worm droppings Diabetic nephropathies sis Glycolipid metabolism |
| 基金项目:四川省中医药管理局科学技术研究专项课题( 2023MS232) |
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| 摘要点击次数: 50 |
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| 中文摘要: |
| 目的基于 AMP活化的蛋白质激酶( AMPK)/沉默信息调节因子 1(SIRT1)-叉头框蛋白 O1(FoxO1)通路探讨蚕沙提取物 deoxynojirimycin(DNJ)对糖尿病小鼠肾纤维化和糖脂代谢的影响。方法 2022年 3月至 2023年 3月，将 12只健康雄性 C57BL/KsJ糖尿病( db/db)小鼠按照随机数字表法分为糖尿病组和 DNJ治疗组( 10 mg·kg.1·d.1，口服， 6周)，各 6只；另外 3只相同条件的非糖尿病 C57野生型小鼠作为对照组(对照组小鼠和糖尿病小鼠口服同等剂量的生理盐水)。监测小鼠血糖、脂质代谢以及肾功能标志物水平。收集小鼠肾组织用于组织学染色，包括苏木精 -伊红( HE)染色，过碘酸希夫( PAS)染色和 Masson三色染色。高糖诱导体外细胞模型并用免疫荧光检测平滑肌肌动蛋白 α(α-SMA)/纤连蛋白( FN)表达评估纤维化，噻唑蓝(MTT)检测细胞活性。另外，蛋白质印迹法检测 AMPK/SIRT1-FoxO1通路水平与自噬水平。结果相比糖尿病小鼠的血糖水(21.79±0.90)mmol/L，经 DNJ治疗 6周后，血糖水平降低至( 15.02±0.61)mmol/L，肾损伤程度得到改善。病理学切片可以观察到平由细胞外基质沉积过多引起的肾纤维化的缓解。与糖尿病小鼠的肾功能指标血尿素氮( 5.24±0.62)mmol/L、血清肌酐(27.68±2.51)mmol/L，尿蛋白 /肌酐( 29.57±2.71)mg/g相比， DNJ治疗组小鼠的肾功能指标降低至血尿素氮( 3.73±0.45)mmol/L、血清肌酐( 14.72±1.66)mmol/L、尿蛋白 /肌酐( 21.49±2.18)mg/g。其次，糖尿病组小鼠脂质代谢中总胆固醇( 3.63±0.07)mmol/L以及甘油三酯( 3.40±0.04)mmol/L也在 DNJ治疗 6周后降低至( 2.54±0.10)mmol/L、(2.01±0.04)mmol/L。在体内和体外实验中，对照组磷酸化 AMPK(p-AMPK)/AMPK、SIRT1、Ac-FoxO1/FoxO1、p62、LC3-Ⅰ/LC3-Ⅱ分别为 1.01±0.02、1.00±0.01、1.01±0.01、 0.99±0.02、0.65±0.08，糖尿病组的 p-AMPK/AMPK、SIRT1和 LC3-Ⅰ/LC3-Ⅱ分别降低至 0.41±0.07、0.52±0.06、0.31±0.09，Ac-FoxO1/FoxO1、p62分别升高至 1.83±0.11和 1.57±0.09；但与糖尿病组相比， DNJ治疗组 p-AMPK/AMPK、SIRT1和 LC3-Ⅰ/LC3-Ⅱ分别逆转至 0.81±0.08、0.84±0.09和 1.79±0.11，Ac-FoxO1/FoxO1、p62分别降低至 0.82±0.09和 1.18±0.11。结论 DNJ治疗通过激活 AMPK促进 SIRT1-FoxO1通路缓解糖尿病小鼠的肾纤维化并降低糖脂代谢水平， AMPK可能是 DNJ在糖尿病中发挥肾脏保护作用的有效靶点。 |
| 英文摘要: |
| Objective To explore the effects of deoxynojirimycin (DNJ) from silkworm sand on renal fibrosis and glycolipid metabo-lism in diabetic mice through the AMPK/SIRT1-FoxO1 pathway.Methods From March 2022 to March 2023, 12 male C57BL/KsJ db/db mice were randomly divided into a diabetic group and a DNJ treatment group (10 mg·kg.1·d.1 oral administration for 6 weeks; 6 mice per group). Three non-diabetic C57 wild-type mice served as controls. Both control and diabetic mice received equivalent volumesof saline. Blood glucose, lipid metabolism, and renal function markers were measured. Kidney tissues were collected for hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and Masson trichrome staining. An in vitro high-glucose model was established, and fibrosis was evaluated via α-SMA/FN immunofluorescence. Cell viability was assessed by MTT assay, while AMPK/SIRT1-FoxO1 pathway proteins and autophagy markers were quantified using Western blotting.Results After 6 weeks of DNJ treatment, blood glucose levels in dia-betic mice decreased significantly from (21.79±0.90) mmol/L to (15.02±0.61) mmol/L. Renal fibrosis improved in DNJ-treated mice, as evidenced by reduced extracellular matrix deposition. The DNJ group showed significant improvements in renal function markers(blood urea nitrogen, serum creatinine, and urine protein/creatinine ratio) and lipid metabolism parameters (total cholesterol and triglyc-erides) compared to the diabetic group (P<0.05). Western blotting revealed that DNJ treatment reversed diabetes-induced alterations in p-AMPK/AMPK, SIRT1, Ac-FoxO1/FoxO1, p62, and LC3-Ⅰ/LC3-Ⅱ ratios in both in vivo and in vitro models (P<0.05). Comparedwith diabetic mice showing blood glucose levels of (21.79±0.90) mmol/L, the DNJ treatment for 6 weeks significantly reduced blood glu-cose levels to (15.02±0.61) mmol/L and improved renal damage. Pathological sections revealed diminished renal fibrosis attributable toexcessive extracellular matrix deposition. Renal function indicators in diabetic mice were recorded as: blood urea nitrogen [(5.24±0.62)mmol/L], serum creatinine [(27.68±2.51) mmol/L], and urine protein/creatinine ratio [(29.57±2.71) mg/g]. Following DNJ treatment,these parameters decreased to (3.73±0.45) mmol/L, (14.72±1.66) mmol/L, and (21.49±2.18) mg/g, respectively. In lipid metabolism, di-abetic mice exhibited total cholesterol [(3.63±0.07) mmol/L] and triglycerides [(3.40±0.04) mmol/L], which decreased to (2.54±0.10)mmol/L and (2.01±0.04) mmol/L, respectively, after DNJ treatment. Both in in vivo and in vitro experiments demonstrated the followingcontrol group ratios: p-AMPK/AMPK (1.01±0.02), SIRT1 (1.00±0.01), Ac-FoxO1/FoxO1 (1.01±0.01), p62 (0.99±0.02), and LC3-Ⅰ/ LC3-Ⅱ (0.65±0.03). In contrast, the diabetic group showed significant decreases in p-AMPK/AMPK (0.41±0.07), SIRT1 (0.52±0.06), and LC3-Ⅰ/LC3-Ⅱ (0.31±0.09), along with increases in Ac-FoxO1/FoxO1 (1.83±0.11) and p62 (1.57±0.09). Notably, DNJ treatment reversed these effects, restoring p-AMPK/AMPK (0.81±0.08), SIRT1 (0.84±0.09), and LC3-Ⅰ/LC3-Ⅱ (1.79±0.11) while reducing Ac-FoxO1/FoxO1 (0.82±0.09) and p62 (1.18±0.11).Conclusions DNJ treatment was found to activate AMPK, thereby promoting the SIRT1-FoxO1 pathway, resulting in alleviating renal fibrosis and reduced glucolipid metabolism levels in diabetic mice. These findingssuggest AMPK as a potential therapeutic target for DNJ-mediated renal protection in diabetes. |
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