氟通过自噬和铁死亡途径诱发肉鸡小肠氧化损伤

doi:10.11843/j.issn.0366-6964.2025.01.040

Abstract

Abstract:

With the continuous development of industrial and agriculture, more and more fluoride enters the production, life and environment through drinking water, medicines, pesticides and fungicides, which may have adverse effects on animals living in the area. In order to explore the related effects of fluoride poisoning on small intestinal injury in broilers, we established an animal experimental model through fluoride diet exposure for subsequent verification. Broilers were divided into control group C (0 mg·kg^-1), low-dose fluoride group L (500 mg·kg^-1), medium dose fluoride group M (1 000 mg·kg^-1) and high-dose fluoride group H (2 000 mg·kg^-1) according to the exposure dose level of sodium fluoride in feed. The effects of fluoride exposure on small intestine of broilers were analyzed. With the increase of sodium fluoride exposure concentration, Nrf2 increased (P < 0.001), Keap1 decreased (P < 0.001), HO-1 decreased (P < 0.001), and NQO1 decreased in the high concentration group (P < 0.01). The Nrf2/HO-1 pathway was activated, but its downstream antioxidant enzymes decreased, suggesting the oxidative stress may occur; the protein expression of p-AKT and p-mTOR increased significantly (P < 0.001), and finally returned to normal or even decreased (P < 0.05), while PI3K decreased significantly (P < 0.001), suggesting that the phosphorylation of PI3K/Akt/mTOR pathway was inhibited; Beclin-1 was significantly increased (P < 0.001), ATG12 was not significantly changed, and LC3 protein was significantly higher than the C group (P < 0.05), indicating the autophagosomes formation and autophagy flow were accelerated, p62 protein combined with autophagy substrate protein (P < 0.001), and the content decreased, indicating the authay pathway was not blocked. At the same time, FTH1 decreased (P < 0.01), NCOA4 increased (P < 0.01), and ferrous ions were released from the degradation of ferritin; ASCL4 and ALOX12 protein increased (P < 0.01), which promoted the lipid peroxidation process; SLC7A11 and GPX4 protein significantly decreased (P < 0.001), and the ability of scavenging lipid peroxide was reduced, which was prone to ferroptosis. Sodium fluoride interfered with the Nrf2 antioxidant pathway, leading to oxidative stress, also activate autophagy and ultimately lead to ferroptosis in the small intestine of broilers.

Key words: sodium fluoride, chicken, small intestine, oxidative stress, autophagy, ferroptosis

CLC Number:

S856.9

WANG Yi, HOU Lulu, FANG Fei, GAO Linying, XIE Shumin, WANG Yu. Fluoride Induced Small Intestine Oxidative Damage in Broilers via Autophagy and Ferroptosis[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 442-454.

Figures/Tables 7

Table 1

Table 2

Fig. 1

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Fig. 5

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基因 Gene	登录号 Accession No.	正向序列(5′→3′) Forward sequence	反向序列(5′→3′) Reverse sequence
β-actin	NM_205518.2	CCGCTCTATGAAGGCTACGC	CTCTCGGCTGTGGTGGTGAA
ACSL4	XM_046917350.1	CCGTTGTTATCTTCTGCGAGACC	ACGCTCCACATTCATTGAGACCATAG
FTH1	NM_205086.2	AGAATGTGAACCAGTCGCTGTTAGAG	TGCTTGATGGCTTTCACCTGCTC
GPX4	NM_204220.3	GCTGTGGAAGTGGCTGAAGGAG	TTGGCCCTCACGGTTGATAAGAAAC
SLC7A11	XM_040670527.2	AACTGCTGGTTATTCGCCCTG	AAGCCAGCAACTACCAGTCCA
AKT	XM_046917866.1	AGGAGGAAGAGATGATGGAT	GAATGGATGCCGTGAGTT
ATG5	XM_046914043.1	GGCACCGACCGATTTAGT	GCTGATGGGTTTGCTTTT
ATG12	XM_046936147.1	GGGACCCTCTATGAGTGTTTTGG	AAGACCACACCTCAGCAACTC
Beclin-1	NM_001006332.1	CGACTGGAGCAGGAAGAAG	TCTGAGCATAACGCATCTGG
mTOR	XM_417614.7	GCTTATGGCACGGTGTTTCC	GCGATCATGCTCTGTTGCAG
p62	XM_040682727.2	GTTAATCCTCTGCGTGTGA	AAGAAGAATCGGTGTGCTAA
NQO1	NM_001277620.2	CGCACCCTGAGAAAACCTCT	TTCTTGAGGGGTCCGGTGAT
HO-1	XM_046921508.1	GTCGTTGGCAAGAAGCATCC	GGGCCTTTTGGGCGATTTTC
Nrf2	XM_046943490.1	CAGGGCAATGCTAGTGTGTACTCATC	AGGGTCTTTCTTTGGTGTGTTCATACG

抗体信息Antibody	稀释率Dilution	来源Source
β-actin	1∶2 000	Abclonal Technology, China
ALOX12	1∶500	Boster, China
GPX4	1∶500	Boster, China
NCOA4	1∶500	Boster, China
FTH1	1∶500	Boster, China
AKT	1∶500	Wanleibio, China
ATG12	1∶500	Wanleibio, China
Beclin-1	1∶500	Wanleibio, China
LC3	1∶500	Wanleibio, China
mTOR	1∶500	Wanleibio, China
PI3K	1∶500	Wanleibio, China
P62	1∶500	Wanleibio, China
Nrf2	1∶500	Wanleibio, China
Keap1	1∶500	Wanleibio, China
NQO1	1∶500	Wanleibio, China
HO-1	1∶500	Wanleibio, China

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Fluoride Induced Small Intestine Oxidative Damage in Broilers via Autophagy and Ferroptosis

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