高硼暴露通过诱导细胞焦亡加剧鸡肝细胞损伤

doi:10.11843/j.issn.0366-6964.2024.10.044

Abstract

Abstract:

We aim to explore the mechanism of boron-induced hepatocytes damage in broilers. Thirty AA broiler chickens were randomly divided into control group (CG), low boron group (LBG) and high boron group (HBG). They were fed with different additional doses of boron in the basal diet: the basal diet (boron 0 mg·kg^-1), low boron diet (boron 120 mg·kg^-1) and high boron diet (boron 240 mg·kg^-1), respectively. Liver tissues were collected 7 and 14 days after treatment. The content of boron in liver tissue was determined by ICP-MS, the ASL and ALT levels were tested by automatic blood biochemistry analyzer, the pathological changes in liver tissue were observed by H & E staining, ultrastructural changes in liver tissue were observed by transmission electron microscopy, and the localized expressions of NLRP3 and IL-1β in liver tissue were detected by immunohistochemistry and immunofluorescence. The mRNA and protein expression levels of NLRP3, Casapse-1, IL-1β, IL-18, and GSDMD were detected by RT-qPCR and Western blot. The results showed that the amount of boron stored in the liver tissue as along dietary boron levels increased, with the duration and dosage of exposure. In addition, the structural integrity of the liver tissue of 14-day-low boron group (14 d-LBG) and 14-day-high boron group (14 d-HBG) was significantly compromised, with an increase in pyroptosis levels with an increase in boron dosage. Compared with CG at the same time, a significant upregulation of mRNA expression levels of NLRP3, Caspase-1 and IL-18 was observed in the 14 d-LBG(P < 0.05). Furthermore, a significant upregulation of both mRNA and protein expression levels of NLRP3, Caspase-1, IL-1β, IL-18 and GSDMD was observed in the 14 d-HBG (P < 0.05). High levels of boron exposure could induce hepatocytes damage in broilers through the pyroptosis pathway.

Key words: boron, pyroptosis, broiler, hepatocyte

CLC Number:

S856.9

Ting HE, Siying LIU, Jinwen QUAN, Weiqian SU, Jiangli HUANG, Yumeng LI, Zhonghua LIU, Wenlan YU. High Levels of Boron Exposure Aggravated Hepatocytes Damage in Broilers via Pyroptosis[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4747-4759.

Figures/Tables 10

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References 32

1	WANG W , XIAO K , ZHENG X T , et al. Effects of supplemental boron on growth performance and meat quality in African ostrich chicks[J]. J Agric Food Chem, 2014, 62 (46): 11024- 11029. doi: 10.1021/jf501789t
2	XIAO K , ANSARI A R , REHMAN Z U , et al. Effect of boric acid supplementation of ostrich water on the expression of Foxn1 in thymus[J]. Histol Histopathol, 2015, 30 (11): 1367- 1378.
3	SCHON M K , BLEVINS D G . Foliar boron applications increase the final number of branches and pods on branches of field-grown soybeans[J]. Plant Physiol, 1990, 92 (3): 602- 607. doi: 10.1104/pp.92.3.602
4	侯宇, 王珂, 杨芸, 等. 喷施硼肥对小麦籽粒产量和面粉品质的影响[J]. 陕西农业科学, 2022, 68 (9): 30-32, 98.
	HOU Y , WANG K , YANG Y , et al. Effect of spaying boron fertilizer on yield and quality of wheat[J]. Shaanxi Journal of Agricultural Sciences, 2022, 68 (9): 30-32, 98.
5	万洋灵, 郭顺堂. 食品中硼的健康风险与控制研究进展[J]. 食品科学, 2016, 37 (21): 265- 270.
	WAN Y L , GUO S T . Progress in understanding and controlling health risks associated with boron in foods[J]. Food Science, 2016, 37 (21): 265- 270.
6	WEI Y N , YANG L , PANDEYA A , et al. Pyroptosis-induced inflammation and tissue damage[J]. J Mol Biol, 2022, 434 (4): 167301. doi: 10.1016/j.jmb.2021.167301
7	VANDE WALLE L , LAMKANFI M . Pyroptosis[J]. Curr Biol, 2016, 26 (13): R568- R572. doi: 10.1016/j.cub.2016.02.019
8	HSU S K , LI C Y , LIN I L , et al. Inflammation-related pyroptosis, a novel programmed cell death pathway, and its crosstalk with immune therapy in cancer treatment[J]. Theranostics, 2021, 11 (18): 8813- 8835. doi: 10.7150/thno.62521
9	关海梅, 曾阳玲, 王恬雯, 等. GSDMD介导的肝细胞焦亡在急性肝衰竭中的致病机制[J]. 海南医学院学报, 2024, 30 (9): 700- 706.
	GUAN H M , ZENG Y L , WANG T W , et al. Pathogenesis of GSDMD-mediated pyrodeath of hepatocytes in acute liver failure[J]. Journal of Hainan Medical University, 2024, 30 (9): 700- 706.
10	周琦, 吕红彬, 王亚平, 等. NLRP3炎症小体在糖尿病视网膜病变神经血管单元损伤中的机制研究[J]. 国际眼科杂志, 2023, 23 (8): 1317- 1322.
	ZHOU Q , LYU H B , WANG Y P , et al. Research on the mechanism of NLRP3 inflammasome in neurovascular unit impairment of diabetic retinopathy[J]. International Eye Science, 2023, 23 (8): 1317- 1322.
11	梁文清, 刘忠华, 常晓月, 等. 长期高铜暴露通过影响线粒体自噬和细胞焦亡诱导大鼠肝组织损伤[J]. 畜牧兽医学报, 2022, 53 (12): 4490- 4500. doi: 10.11843/j.issn.0366-6964.2022.12.034
	LIANG W Q , LIU Z H , CHANG X Y , et al. Long-term exposure to high levels of copper induced liver injury in rats by affecting mitophagy and pyroptosis[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53 (12): 4490- 4500. doi: 10.11843/j.issn.0366-6964.2022.12.034
12	WAN F , ZHONG G L , WU S F , et al. Arsenic and antimony co-induced nephrotoxicity via autophagy and pyroptosis through ROS-mediated pathway in vivo and in vitro[J]. Ecotoxicol Environ Saf, 2021, 221, 112442. doi: 10.1016/j.ecoenv.2021.112442
13	LI Y X , ZHONG G L , HE T , et al. Effect of arsenic and copper in kidney of mice: Crosstalk between Nrf2/ Keap1 pathway in apoptosis and pyroptosis[J]. Ecotoxicol Environ Saf, 2023, 266, 115542. doi: 10.1016/j.ecoenv.2023.115542
14	WEIR JR R J , FISHER R S . Toxicologic studies on borax and boric acid[J]. Toxicol Appl Pharmacol, 1972, 23 (3): 351- 364. doi: 10.1016/0041-008X(72)90037-3
15	王靳琎, 李雪莹, 易进科, 等. 硼酸钠对小鼠的急性毒性实验研究[J]. 中国医药科学, 2023, 13 (9): 22-25, 66.
	WANG J J , LI X Y , YI J K , et al. Experimental study on the acute toxicity of sodium borate to mice[J]. China Medicine and Pharmacy, 2023, 13 (9): 22-25, 66.
16	王行智, 张文, 陈婷, 等. 微波消解ICP-MS/MS测定畜禽表皮组织17种无机元素[J]. 食品工业, 2023, 44 (2): 272- 276.
	WANG X Z , ZHANG W , CHEN T , et al. Determination of 17 inorganic elements in livestock and poultry epidermal tissue by microwave ICP-MS/MS[J]. The Food Industry, 2023, 44 (2): 272- 276.
17	尹玉霞. 硼泥的环境问题及资源化利用[J]. 中国资源综合利用, 2020, 38 (2): 72- 75.
	YIN Y X . Environmental problems and comprehensive utillization of boron mud[J]. China Resources Comprehensive Utilization, 2020, 38 (2): 72- 75.
18	杜鹏, 韩京秀, 董少霞, 等. 硼的健康风险研究进展[J]. 环境卫生学杂志, 2015, 5 (2): 177- 183.
	DU P , HAN J X , DONG S X , et al. Research progress in health risks of boron[J]. Journal of Environmental Hygiene, 2015, 5 (2): 177- 183.
19	高小净, 孙爱德, 张旭, 等. 硼肥施用对辣椒果实中硼累积效果的影响[J]. 安徽农学通报, 2023, 29 (18): 17- 20.
	GAO X J , SUN A D , ZHANG X , et al. Effect of borax application on boron accumulation in capsicum fruit[J]. Anhui Agricultural Science Bulletin, 2023, 29 (18): 17- 20.
20	赵春芳, 金吉雄, 张梦玲, 等. 硼上调皖西白鹅产蛋期卵巢IGFBP3、DRD2和CDK1基因表达[J]. 安徽科技学院学报, 2022, 36 (1): 1- 6.
	ZHAO C F , JIN J X , ZHANG M L , et al. Effect of boron on gene expression of IGFBP3, DRD2 and CDK1 in ovary of Wanxi white geese during laying period[J]. Journal of Anhui Science and Technology University, 2022, 36 (1): 1- 6.
21	赵春芳, 陈宁, 张路, 等. 硼对皖西白鹅产蛋期下丘脑中GRB2、HSP70和AMPK基因表达的影响[J]. 皖西学院学报, 2022, 38 (2): 8- 12.
	ZHAO C F , CHEN N , ZHANG L , et al. Effect of boron on of GRB2, HSP70 and AMPK mRNA expression in the hypothalamus of Wanxi white geese during laying period[J]. Journal of West Anhui University, 2022, 38 (2): 8- 12.
22	丁永旺, 任曼, 赵春芳, 等. 硼对大鼠肝脏显微结构、肝糖原含量、抗氧化功能及细胞增殖和凋亡相关基因表达的影响[J]. 动物营养学报, 2022, 34 (3): 1963- 1974.
	DING Y W , REN M , ZHAO C F , et al. Effects of boron on microstructure, hepatic glycogen content, antioxidant function, cell proliferation and apoptosis related gene expression in liver of rats[J]. Chinese Journal of Animal Nutrition, 2022, 34 (3): 1963- 1974.
23	ZHANG Q , HE X , YU Q , et al. Endoplasmic reticulum stress regulates pyroptosis in BPDE-induced BEAS-2B cells[J]. Environ Toxicol, 2022, 37 (7): 1768- 1780.
24	雷倩, 易焘, 陈灿. 细胞焦亡效应机制及相关疾病研究进展[J]. 医学研究生学报, 2017, 30 (6): 648- 652.
	LEI Q , YI T , CHEN C . Research progresses of the effective mechanism of pyroptosis and its related diseases[J]. Journal of Medical Postgraduates, 2017, 30 (6): 648- 652.
25	MCKENZIE B A , MAMIK M K , SAITO L B , et al. Caspase-1 inhibition prevents glial inflammasome activation and pyroptosis in models of multiple sclerosis[J]. Proc Natl Acad Sci U S A, 2018, 115 (26): E6065- E6074.
26	DE CARVALHO RIBEIRO M , SZABO G . Role of the inflammasome in liver disease[J]. Annu Rev Pathol, 2022, 17, 345- 365.
27	CHANG Y , ZHU J , WANG D , et al. NLRP3 inflammasome-mediated microglial pyroptosis is critically involved in the development of post-cardiac arrest brain injury[J]. J Neuroinflammation, 2020, 17 (1): 219.
28	ZHANG Z H , BAI H , MA X Y , et al. Blockade of the NLRP3/caspase-1 axis attenuates ketamine-induced hippocampus pyroptosis and cognitive impairment in neonatal rats[J]. J Neuroinflammation, 2021, 18 (1): 239.
29	JU J , LIU Y Y , LIANG H H , et al. The role of pyroptosis in endothelial dysfunction induced by diseases[J]. Front Immunol, 2023, 13, 1093985.
30	ZHANG C Y , LIN T J , NIE G H , et al. Cadmium and molybdenum co-induce pyroptosis via ROS/PTEN/PI3K/AKT axis in duck renal tubular epithelial cells[J]. Environ Pollut, 2021, 272, 116403.
31	HENAO-MEJIA J , ELINAV E , THAISS C A , et al. Inflammasomes and metabolic disease[J]. Annu Rev Physiol, 2014, 76 (1): 57- 78.
32	廖建昭, 杨帆, 裴若男, 等. 铜通过活性氧诱导鸡肝细胞发生焦亡[J]. 中国兽医学报, 2018, 38 (12): 2374- 2378.
	LIAO J Z , YANG F , PEI R N , et al. Copper induces pyroptosis by generating ROS in chicken hepatocyte[J]. Chinese Journal of Veterinary Science, 2018, 38 (12): 2374- 2378.

日粮组成Diet composition		营养水平Nutrients levels
配料Ingredients	含量Content	指标Indices	水平Levels
玉米Conrn	54.82	干物质Dry matter	88.60
豆粕Soybean	37.77	粗蛋白Crude protein	23.00
豆油Soybean oil	2.50	代谢能/(MJ·kg^-1)	12.70
石粉Limestone	1.01	钙Calcium	0.95
磷酸氢钙Dicalcium phosphate	1.75	有效磷Avail-P	0.46
食盐Salt	0.40
添加剂^* Additive^*	0.50
DL-蛋氨酸DL-Methionine	0.25
玉米淀粉+铜Conrn starch+Cu	1.00

基因 Genes	引物序列(5′→3′) Primer sequence	产物长度/bp Product length	序列号 Sequence number
NLRP3	AGCTACCACACATCTAGGAT	207	XM_046918112.1
	GGTGTCCAAATCCTCAATCT
Caspase-1	TTCCTTCAACACCATCTACG	209	XM_040687588.2
	GGTGAGCTTCTCTGGTTTTA
IL-1β	CAGCCTCAGCGAAGAGACCTT	165	XM_046931582.1
	ACTGTGGTGTGCTCAGAATCC
IL-18	GCAGTACGGCTTAGAGAAA	198	XM_046932263.1
	GTACATTCCACTGCCAGATT
GADPH	GCCATCACAGCCACACAGAAGA	206	NM_204305.2
	CGGCAGGTCAGGTCAACAACAG
GSDMD	CTGCTCTGATCTACAGGTTGTTCCG	79	XM_046911140
	GCTCAGTGCCAAGGTGCCATC

组别Group	AST	ALT
7 d-对照组7 d-CG	1.600±0.100^a	57.625±8.922^a
7 d-低硼组7 d-LBG	2.250±0.208^a	139.925±15.897^b
7 d-高硼组7 d-HBG	3.100±0.200^b	206.825±1.053^b
14 d-对照组14 d-CG	1.650±0.250^a	52.633±14.836^a
14 d-低硼组14 d-LBG	2.575±0.171^b	230.725±17.814^b
14 d-高硼组14 d-HBG	3.967±1.159^b	282.575±23.517^b

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High Levels of Boron Exposure Aggravated Hepatocytes Damage in Broilers via Pyroptosis

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