Nutr Res Pract.  2024 Jun;18(3):325-344. 10.4162/nrp.2024.18.3.325.

Kaempferol ameliorates metabolic syndrome by inhibiting inflammation and oxidative stress in high-fat diet-induced obese mice

• Shin SK
• Kwon EY

• KMID: 2558507
• DOI: http://doi.org/10.4162/nrp.2024.18.3.325

Abstract

BACKGROUND/OBJECTIVES
Kaempferol (Ka) is one of the most widely occurring flavonoids found in large amounts in various plants. Ka has anti-obesity, antioxidant, and antiinflammatory effects. Despite the numerous papers documenting the efficacy of Ka, some controversy remains. Therefore, this study examined the impact of Ka using 3T3-L1 and highfat diet-induced obese mice.
MATERIALS/METHODS
3T3-L1 cells were treated with 50 μM Ka from the initiation of 3T3-L1 differentiation at D0 until the completion of differentiation on D8. Thirty male mice (C57BL/6J, 4 weeks old) were divided into 3 groups: normal diet (ND), high-fat diet (HFD), and HFD + 0.02% (w/w) Ka (Ka) group. All mice were fed their respective diets ad libitum for 16 weeks. The mice were sacrificed, and the plasma and hepatic lipid levels, white adipose tissue weight, hepatic glucose level, lipid level, and antioxidant enzyme activities were analyzed, and immunohistochemistry staining was performed.
RESULTS
Ka suppressed the hypertrophy of 3T3-L1 cells, and the Ka-supplemented mice showed a significant decrease in perirenal, retroperitoneal, mesenteric, and subcutaneous fat compared to the HFD group. Ka supplementation in high-fat diet-induced obese mice also improved the overall blood lipid concentration (total cholesterol, non-high-density lipoprotein-cholesterol, phospholipids, and apolipoprotein B). Ka supplementation in highfat-induced obesity mice reduced hepatic steatosis and insulin resistance by modulating the hepatic lipid (glucose-6-phosphate dehydrogenase, fatty acid synthase, malic enzyme, phosphatidate phosphohydrolase, and β-oxidation) activities and glucose (glucokinase, phosphoenolpyruvate carboxykinase, and G6pase)-regulating enzymes. Ka supplementation ameliorated the erythrocyte and hepatic mitochondrial H₂O₂ and inflammation levels (plasma tumor necrosis factor-alpha, monocyte chemoattractant protein-1, interleukin-6, and interferon-gamma and fibrosis of liver and epididymal fat).
CONCLUSION
Ka may be beneficial for preventing diet-induced obesity, inflammation, oxidative stress, and diabetes.

Keyword

Obesity; inflammation; oxidative stress; fatty liver; metabolic syndrome

Figure

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  Fig. 1 Effects of Ka on the cell viability of 3T3-L1 cells (A), ORO staining of 3T3-L1 cells treated with Ka 50 μM (B), mRNA expression of lipogenesis of 3T3-L1 cells treated with Ka 50 μM (C), mRNA expression of the inflammatory factors of 3T3-L1 cells treated with Ka 50 μM (D). Data are expressed as the mean ± SE of the mean. DMSO, dimethyl sulfoxide; Ka, kaempferol; ORO, Oil Red O; Pparγ, peroxisome proliferator activated receptor gamma; FAS, fatty acid synthase; TNFα, tumor necrosis factor-alpha;Cd44, CD44 molecules; Mcp-1, monocyte chemoattractant protein-1. Student’s t-test, n = 3, *P < 0.05, **P < 0.01, ***P < 0.001 vs. DMSO 0.1% (CON).

• 

  Fig. 2 Effects of Ka on body weight (A), food intake; FER (B), WAT weight (C), representative images for H&E staining of epididymal WAT (D), plasma lipid profiles (E) in C57BL/6J mice fed a HFD; The data are expressed as the mean ± SE of the mean. ND, normal diet (AIN-76); Ka, kaempferol; HFD, high-fat diet; HFD + Ka, high-fat diet + 0.02% kaempferol; FER, food efficiency ratio; WAT, white adipose tissue; H&E, hematoxylin and eosin; TC, total cholesterol; HDL-C, high-density lipoprotein cholesterol; HTR, high-density lipoprotein cholesterol/total cholesterol ratio; TG, triglycerides; FFA, free fatty acid; Apo, apolipoprotein; RBC, red blood cell; TBARS, thiobarbituric acid reactive substances. Student’s t-test, *P < 0.05, ***P < 0.001 vs. ND; #P < 0.05, ###P < 0.001 vs. HFD.

• 

  Fig. 3 Effects of Ka on the liver weight (A), representative images of H&E staining of the liver (B), hepatic lipid profiles (C), hepatic lipid-regulating enzyme activities (D), hepatic cholesterol-regulating enzyme activities (E), and hepatic TBARS (F) in C57BL/6J mice fed a HFD; the data are expressed as the mean ± SE of the mean. ND, normal diet (AIN-76); HFD, high-fat diet; Ka, kaempferol; HFD + Ka, high-fat diet + 0.02% kaempferol; H&E, hematoxylin and eosin; TG, triglyceride; FA, fatty acid; FAS, fatty acid synthase; ME, malic enzyme; G6PD, glucose-6-phosphate dehydrogenase; PAP, phosphatidate phosphohydrolase; CPT, carnitine palmitoyltransferase; HMGCR, 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase; ACAT, acyl-coenzyme A:cholesterol acyltransferase; TBARS, thiobarbituric acid reactive substances; MDA, malondialdehyde. Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001 vs. ND; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. HFD.

• 

  Fig. 4 Effects of Ka on the plasma glucose, insulin, glucagon and HOMA-IR (A), IPGTT (B), hepatic glycogen and glucose-regulating enzymes activities (C), representative images for insulin and glucagon of the pancreas (D) in C57BL/6J mice fed a HFD; The data are expressed as the mean ± SE of the mean.ND, normal diet (AIN-76); HFD, high-fat diet; Ka, kaempferol; HFD + Ka, high-fat diet + 0.02% kaempferol; HOMA-IR, homeostatic model assessment of insulin resistance; AUC, area under curve; GK, glucokinase; PEPCK, phosphoenolpyruvate carboxykinase; IPGTT, intraperitoneal glucose tolerance test. Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001 vs. ND; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. HFD.

• 

  Fig. 5 Effects of Ka on adipokines and hormones (A), plasma inflammatory factors (B), representative images of CD68, MCP-1, MT staining of liver (C) and epididymal WAT (D) in C57BL/6J mice fed a HFD; The data are expressed as the mean ± SE of the mean. ND, normal diet (AIN-76); HFD, high-fat diet; Ka, kaempferol; HFD + Ka, high-fat diet + 0.02% kaempferol; PAI-1, plasminogen activator inhibitor-1; GIP, gastric inhibitory polypeptide; TNFα, tumor necrosis factor-alpha; MCP-1, monocyte chemoattractant protein-1; IL, interleukin; IFN-γ, interferon-gamma; CD, CD molecules; MT, Masson’s trichrome; WAT, white adipose tissue. Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001 vs. ND; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. HFD.

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