柠檬苦素通过激活AMPK减轻高脂饲料诱导的小鼠肾损伤

doi:10.11843/j.issn.0366-6964.2023.04.037

摘要/Abstract

摘要： 旨在研究柠檬苦素（Limonin，LM）对高脂饲料诱导的小鼠肾损伤的保护作用。选用体重21~23 g雄性C57BL/6小鼠25只，随机分为5组（n=5）：对照组、LM组、高脂组、高脂+低剂量LM组（60 mg·（kg·d）^-1）和高脂+高剂量LM组（120 mg·（kg·d）^-1）。12周后称量各组小鼠体重；使用自动生化分析仪检测血清甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白（HDL）、低密度脂蛋白（LDL）、肌酐（Cr）、尿素氮（BUN）；使用试剂盒检测肾组织TG、TC；通过苏木精-伊红（HE）、糖原（PAS）染色观察肾组织病理组织学变化；通过免疫组织化学检测肾中脂肪酸合成酶（FAS）、甾醇调节元件结合蛋白1（SREBP1）、脂肪甘油三酯脂肪酶（ATGL）、激素敏感性脂肪酶（HSL）蛋白含量；通过蛋白免疫印迹检测AMP活化蛋白激酶（AMPK）、磷酸化的AMPK （P-AMPK）、FAS、SREBP1、ATGL、HSL及肾损伤分子1（KIM1）的表达；通过分子对接检验LM、阿卡地新（AICA Riboside，AICAR）以及多索吗啡（Dorsomorphin，Compound C）与AMPK蛋白结合能力。结果显示，与高脂组相比，LM干预显著降低了小鼠血清和肾中甘油三酯含量（P<0.05），改善了高脂饲喂引起的肾组织病理学损伤，促进AMPK磷酸化（P<0.05），显著上调ATGL、HSL蛋白的表达（P<0.05），抑制SREBP1、FAS蛋白的表达（P<0.05）。分子对接结果显示LM能与AMPK紧密结合且存在多个与AICAR结合AMPK的相同位点。结果提示，LM通过激活AMPK减轻肾脂肪异位沉积从而改善高脂饲料诱导的小鼠肾损伤。

关键词: 柠檬苦素, 高脂饲料, 肾损伤, AMP活化蛋白激酶

Abstract: The study aimed to investigate the protective effect of Limonin (LM) on kidney injury induced by high-fat diet in mice. Twenty-five male C57BL/6 mice weighed 21-23 g were selected and randomly divided into five groups (n=5) according to the test requirements:control group, LM group, high-fat group, high-fat + low-dose LM group (60 mg·(kg·d)^-1) and high-fat+high-dose LM group (120 mg·(kg·d)^-1). After 12 weeks the body weight of mice were detected. The automatic biochemical analyzer was used to measure triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), creatinine (Cr), and urea nitrogen (BUN) in serum. The kits were used to measure TC, TG in the kidney. Hematoxylin-eosin (HE) and periodic acid-schiff (PAS) staining were performed to observe histological changes of kidney tissues. The protein expression of fatty acid synthase (FAS), sterol regulatory element binding protein 1 (SREBP1), adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL) in the kidney were measured by immunohistochemistry. The protein expression of AMP-activated protein kinase (AMPK), phosphorylated AMPK (P-AMPK), FAS, SREBP1, ATGL, HSL and kidney injury molecule 1 (KIM1) was measured by immunoblot. The bound ability of LM、AICA Riboside (AICAR) and Dorsomorphin(Compound C) with AMPK was examined by molecular docking. The results showed that compared with high-fat group, LM intervention significantly reduced TG content in serum and kidney (P<0.05), improved histopathological kidney injury in mice fed with high-fat diet, promoted AMPK phosphorylation (P<0.05), significantly up-regulated the protein expression of ATGL and HSL (P<0.05), and inhibited the protein expression of SREBP1 and FAS (P<0.05). In addition, molecular docking showed that LM could bind tightly to AMPK proteins and had multiple sites identical to those of AICAR binding AMPK. The results suggest that LM attenuates kidney fat ectopic deposition and improves the high-fat diet-incluced kidney injury in mice by activating AMPK.

Key words: Limonin, high-fat diet, kidney injury, AMP-activated protein kinase

中图分类号:

S854

聂桐, 马隽. 柠檬苦素通过激活AMPK减轻高脂饲料诱导的小鼠肾损伤[J]. 畜牧兽医学报, 2023, 54(4): 1742-1750.

NIE Tong, MA Jun. Limonin Attenuates High-fat Diet-induced Kidney Injury in Mice by Activating AMPK[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1742-1750.

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