Alleviating Effect of Astaxanthin on Liver Injury Induced by Aflatoxin B 1 and Its Mechanism

doi:10.13560/j.cnki.biotech.bull.1985.2025-0199

Abstract

Abstract:

Objective To investigate the protective effects and molecular mechanisms of astaxanthin (AST) on aflatoxin B1 (AFB1)-induced liver injury. Method AFB1-induced mouse hepatic parenchymal AML21 cells and C57BL/6 mouse models were constructed, and commercial kits were used to detect the levels of biochemical markers (aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase), oxidative markers (ROS, MDA, SOD, GSH, and CAT), and inflammatory markers (IL-1β and IL-18) after AST treatment. The impact of AST on the Nrf2 and pyroptosis signaling pathways was detected using the Western Blot (WB) method. Result Both in vivo and in vitro results consistently indicate that AST effectively alleviated the abnormalities in biochemical markers, oxidative markers, and inflammatory markers caused by AFB1. By activating the Nrf2 signaling pathway, AST significantly increased the levels of antioxidant proteins such as NQO1, HO-1, GCLC, and GCLM. However, the protein expression s of Nrf2, NQO1, and HO-1significantly reduced when AML21 cells were treated with the Nrf2 inhibitor ML385. But the protein expressions significantly reversed when AST and ML385 were used in combination. Additionally, AST inhibited the pyroptosis pathway, significantly decreasing the protein expressions of NLRP3, ASC, Caspase-1, and GSDMD. Conclusion AST alleviates AFB1-induced liver injury by activating the Nrf2 signaling pathway and inhibiting cell pyroptosis, thereby resisting oxidative stress and inflammatory responses.

Key words: astaxanthin, aflatoxin B₁, liver injury, oxidative stress, inflammation, pyroptosis

YANG Wei, GUAN Hai-feng, REN Xin-hui, PENG Jin-ju, CHEN Zhi-bao. Alleviating Effect of Astaxanthin on Liver Injury Induced by Aflatoxin B ₁ and Its Mechanism[J]. Biotechnology Bulletin, 2025, 41(10): 334-342.

Figures/Tables 8

Fig. 1 Effect of AST on AFB1-induced AML21 cell damageA-C: Biochemical indicators. D-F: Antioxidation indexes. G-H: Inflammatory factors. I-J: ROS levels and relative quantitative analysis. Compared with CON, #P<0.05; ##P<0.01; ###P<0.001. Compared with AFB1, *P<0.05; **P<0.01; ***P<0.001, the same below

Fig. 2 Effects of AST on Nrf2 pathway in AFB1-induced AML21 cellsA-B: The results of AST on the protein expressions and relative protein quantitative analysis of Nrf2, Keap1, NQO1, HO-1, GCLC and GCLM in AFB1-induced AML21 cells. C-D: The results of AST and Nrf2 inhibitor ML385 on the protein expressions and relative protein quantitative analysis of Nrf2, NQO1, and HO-1 in AFB1-induced AML21 cells

Fig. 3 Effect of AST on pyroptosis pathways in AFB1-induced AML21 cellsA-B: The results of AST on the protein expressions and relative protein quantitative analysis of NLRP3, ASC, GSDMD and Caspase-1 in AFB1-induced AML21 cells

Fig. 4 Effect of AST on the liver pathological injury in AFB1-induced miceTop A: Observation of liver injury in mice, black arrows indicate the occurrence of hepatocyte swelling with vacuolar degeneration. Bottom A: HE staining of mouse liver tissue (400×)

Table 1 Analysis of live function index in liver tissue and serum

组织 Tissue	处理 Treatment	谷草转氨酶 Glutamic oxalacetic transaminase（U/L）	谷丙转氨酶 Glutamic-pyruvic transaminase（U/L）	碱性磷酸酶 Alkaline phosphatase（U/L）
肝脏 Liver	CON	246.828±0.612	123.229±4.075	20.459±3.149
	AFB₁	405.961±20.85^##	225.165±7.041^##	36.847±0.339^##
	AST30	307.242±3.669^**	150.364±0.654^**	24.613±2.333^**
	AST100	300.681±1.447^**	143.950±6.307^**	18.752±1.353^**
	DDB30	330.726±4.687^**	155.653±2.001^**	27.069±0.352^**
血清 Serum	CON	16.944±0.307	22.708±0.676	30.801±0.408
	AFB₁	35.599±1.417^##	62.137±0.378^##	74.568±1.569^##
	AST30	16.465±0.109^**	32.139±0.577^**	46.452±1.645^**
	AST100	11.187±0.392^**	21.456±1.185^**	31.701±4.681^**
	DDB30	20.953±0.539^*	23.942±0.769^**	36.880±1.827^**

Fig. 5 Effects of AST on the oxidative stress and inflammation in AFB1-induced miceA-D: Effect of AST on oxidative indices in AFB1-induced mouse liver tissue. E-H: Effect of AST on antioxidative markers in the serum of AFB1-induced mice. I-J: Effect of AST on inflammatory factors in the serum of AFB1-induced mice

Fig. 6 Effect of AST on AFB1 induced Nrf2 pathway in mice liver tissueA-B: The results of AST on the protein expressions and relative protein quantitative analysis of Nrf2, Keap1, NQO1, HO-1, GCLC and GCLM in AFB1-induced mouse liver tissue

Fig. 7 Effect of AST on AFB1 induced pyroptosis pathway in mice liver tissueA-B: The results of AST on the protein expressions and relative protein quantitative analysis of NLRP3, ASC, GSDMD and Caspase-1 in AFB1-induced mouse liver tissue

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Alleviating Effect of Astaxanthin on Liver Injury Induced by Aflatoxin B ₁ and Its Mechanism

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