Kaempferol attenuates diet-induced obesity and hepatic steatosis in C57BL/6J mice fed an Indian diet-mimicking regimen

Figure 1 Optimization of in-house prepared Indian high fat diet in C57BL/6J mice. A: Change in body weight of C57BL/6J mice after consuming the in-house formulated Indian high fat diet (HFD); B: The concentration of transforming growth factor beta 1. Data represented in mean ± SD (n = 3), ^aP < 0.05 (control vs Indian HFD); C: Representative images of hematoxylin and eosin-stained liver sections of mice fed with normal pellet diet and Indian HFD after 8.5 weeks of induction. All photomicrographs: Hematoxylin and eosin stain; magnification 40 × (scale bar = 100 μm); D: Non-alcoholic fatty liver disease activity score conducted in liver samples of mice fed with normal pellet and Indian HFD group. Data represented in mean ± SD (n = 3), ^aP < 0.001 (control, micro-vesicular steatosis vs Indian HFD, micro-vesicular steatosis); ^bP < 0.001 (control, macro-vesicular steatosis vs Indian HFD, macro-vesicular steatosis); ^cP < 0.001 (control, lobular inflammation vs Indian HFD, lobular inflammation). HF: High fat; HFD: High fat diet; NAFLD: Non-alcoholic fatty liver disease; TGF-β1: Transforming growth factor beta 1.

Figure 2 Effect of kaempferol in Indian high fat diet model induced metabolic dysfunction-associated steatotic liver disease. A: Weekly body weight changes in mice fed the Indian high fat diet (HFD) for 8.5 weeks, followed by treatment with kaempferol or vitamin E on alternate days for 4 consecutive weeks. The kaempferol-treated group (blue line) demonstrates a marked reduction in body weight compared to the positive control group (green line); B: Non-alcoholic fatty liver disease activity score conducted in liver samples from control group, Indian HFD group, kaempferol group and vitamin E group confirms the development of metabolic dysfunction-associated steatotic liver disease in Indian HFD group and its alleviation in kaempferol treatment group; C-F: The histology images from control group, Indian HFD group, kaempferol group and vitamin E groups. All photomicrographs: Hematoxylin and eosin stain; magnification 40 × (scale bar = 100 μm). All data’s represented in mean ± SD (n = 3), ^aP < 0.001 (control, micro-vesicular steatosis vs Indian HFD, micro-vesicular steatosis); ^bP < 0.0001 (control, macro-vesicular steatosis vs Indian HFD, macro-vesicular steatosis); ^cP < 0.0001 (control, lobular inflammation vs Indian HFD, lobular inflammation); ^dP < 0.001 (control, hepatocellular hypertrophy vs Indian HFD, hepatocellular hypertrophy); ^eP < 0.0001 (Indian HFD, macro-vesicular steatosis vs kaempferol-treated, macro-vesicular steatosis); ^fP < 0.001 (Indian HFD, lobular inflammation vs kaempferol-treated, lobular inflammation); ^gP < 0.01 (Indian HFD, hepatocellular hypertrophy vs kaempferol-treated, hepatocellular hypertrophy). KMF: Kaempferol; Vit: Vitamin E.

Figure 3 Kaempferol alleviates the liver function parameters and triglyceride levels in an ex-vivo precision-cut liver slice model. A: For alanine aminotransferase, data represented in mean ± SD (n = 3), ^aP < 0.0001 [control, vs palmitic acid (PA)-treated, 240 μM]; ^bP < 0.0001 (PA-treated, 240 μM vs kaempferol, 25 μM); ^cP < 0.0001 (PA-treated, 240 μM vs kaempferol, 50 μM); B: For aspartate aminotransferase, data represented in mean ± SD (n = 3), ^aP < 0.001 [control vs PA-treated, 240 μM]; ^bP < 0.001 (PA-treated, 240 μM vs kaempferol, 25 μM); ^cP < 0.001 (PA-treated, 240 μM vs kaempferol, 50 μM); C: For alkaline phosphatase, data represented in mean ± SD (n = 3), ^aP < 0.05 [control vs PA-treated, 240 μM]; ^bP < 0.05 (PA-treated, 240 μM vs kaempferol, 25 μM); ^cP < 0.05 (PA-treated, 240 μM vs kaempferol, 50 μM); D: For triglycerides, data represented in mean ± SD (n = 3), ^aP < 0.001 [control vs PA-treated, 240 μM]; ^bP < 0.001 (PA-treated, 240 μM vs kaempferol, 25 μM); ^cP < 0.001 (PA-treated, 240 μM vs kaempferol, 50 μM); ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; PA: Palmitic acid; KMF: Kaempferol.