TBHQ对鸡舍PM2.5诱导鸡胚肺组织细胞焦亡、坏死和炎症损伤的影响

doi:10.3864/j.issn.0578-1752.2023.04.015

Abstract

Abstract:

【Objective】 The aim of this study was to investigate the protective effect and mechanism of tert-butylhydroquinone (TBHQ) on chicken embryo lung injury induced by fine particulate matter (PM_2.5) in chicken house, so as to provide a theoretical basis for prevention and mitigation of chicken respiratory health problems caused by PM_2.5 pollution in chicken house. 【Method】 14-day-old chicken embryos were selected as the research model in this study. Firstly, a model of lung tissue damage induced by PM_2.5 from chicken houses was established, and then the alleviating effects of TBHQ on lung injury of chicken embryos was studied. In the experiment of establishing the model of chicken embryo lung tissue injury induced by PM_2.5 from chicken houses, different concentrations of PM_2.5 (0, 0.25, 0.5, and 1 mg·mL^-1) were injected into egg albumens. After 5 days, the survival rate and lung tissues of chicken embryos were observed, and the appropriate concentration of PM_2.5 (0.25 mg·mL^-1) was selected to establish the model of chicken embryo lung injury. In the experiment of the effects of TBHQ on PM_2.5-induced lung injury of chicken embryos, chicken embryos were divided into control group (saline), PM_2.5 group (0.25 mg·mL^-1 PM_2.5), TBHQ group (0.1 μg·mL^-1 TBHQ) and PM_2.5+TBHQ group (0.25 mg·mL^-1 PM_2.5+0.1 μg·mL^-1 TBHQ). The experiment lasted for 5 days. The lung tissues were collected and the egg weight, embryo weight and lung tissue weight were recorded. The expressions of MDA, T-SOD, T-AOC and pyroptosis-related genes (NLRP3, Caspase-1, IL-18, IL-1β) and necroptosis-related genes (RIPK1, RIPK3, MLKL) in lung tissues of chicken embryos were detected. 【Result】 The results suggested that in the experiment to establish a model for PM_2.5-induced lung tissue damage in chicken embryos, different concentrations of PM_2.5 (0, 25, 50, and 100 μg·mL^-1) from chicken houses did not significantly affect the survival of chicken embryos, but the lung tissues of chicken embryos in the 0.25 mg·mL^-1 PM_2.5 group showed inflammatory cell infiltration. The lung tissues of chicken embryos in 0.5 mg·mL^-1 and 1 mg·mL^-1 PM_2.5 groups showed pulmonary edema. In the experiment on the effects of TBHQ on PM_2.5-induced lung damage in chicken embryos, there was no effect on embryo weight, lung tissue weight and related indices in all groups. Compared with the PM_2.5 group, the inflammatory cell infiltration and MDA levels were reduced in the lung tissues of chicken embryos co-treated with TBHQ and PM_2.5. RT-PCR results showed that the expression of the Caspase-1 gene in the TBHQ group was significantly increased compared with the control group (P<0.01), as were the expressions of IL-1β (P<0.01) and RIPK1 (P<0.01) genes in the PM_2.5 group. Compared with the PM_2.5 group, the expressions of IL-18 (P<0.01), IL-1β (P<0.01), RIPK1 (P<0.01), and MLKL (P<0.01) genes were significantly down in the TBHQ and PM_2.5 co-treatment groups. On the contrary, the expressions of Caspase-1 (P<0.01), RIPK3 (P<0.05) genes in the TBHQ and PM_2.5 co-treatment groups were significantly increased compared with the PM_2.5 group. 【Conclusion】TBHQ could alleviate inflammatory damage in chicken embryo lung tissues induced by PM_2.5 from chicken houses by inhibiting oxidative stress and reducing the gene expressions of pyroptosis and necroptosis.

Key words: TBHQ, pyroptopsis, necroptosis, PM_2.5, chicken embryos, oxidative stress

XI MengXue, SHEN Dan, SHI YiFan, LI ChunMei. Effects of TBHQ on Pyroptosis, Necroptosis and Inflammatory Damage of Chicken Embryonic Lung Tissues Induced by PM_2.5 from Chicken Houses[J].Scientia Agricultura Sinica, 2023, 56(4): 779-787.

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References 30

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引物名称 Gene	引物序列 Sequence (5′ to 3′)
Caspase-1	TTCCTTCAACACCATCTACG / GGTGAGCTTCTCTGGTTTTA
IL-1β	ACTGGGCATCAAGGGCTACA / GCTGTCCAGGCGGTAGAAGA
IL-18	AGTTGCTTGTGGTTCGTCCA / TCCACTGCCAGATTTCACCTC
NLRP3	AGCTACCACACATCTAGGAT / GGTGTCCAAATCCTCAATCT
PERK	TCGAGCTGCTTTACCCTTTC / CTCATTGTCCGTGACCTCTG
CHOP	CAGGAAGAAGAGCTGGCCCCACT / TGCTGTGCTCGCCGTGCTGT
IRE1	CCCAAAGCATCAAACCATTC / CAACGTCGCGGTTATCAAAT
ATF6	CGTCGTCTGAACCACTTACTGA / CCTTCTTTCCTAACAGCCACAC
GRP78	GAATCGGCTAACACCAGAGGA / CGCATAGCTCTCCAGCTCATT
RIPK1	CTTCAACCAGCGCCATTAGC / TTGAGTCTTCTGTATCCGTGTCT
RIPK3	ACATCCTTCGCTCACAGCAA / ACCTGTGCTGCCTTCTCTCC
MLKL	AAGGTGGACTGGATGCAAGG / TAGAGGTCGTAGCGCTCAGT
β-actin	TGTTACCAACACCCACACCC / TCCTGAGTCAAGCGCCAAAA

浓度 Concentration (mg·mL^-1)	胚数Embryo number		总数 Total	鸡胚存活率 Survival rate of chicken embryo (%)
浓度 Concentration (mg·mL^-1)	死亡Death	存活Survival	总数 Total	鸡胚存活率 Survival rate of chicken embryo (%)
0	0	5	5	100
0.25	0	5	5	100
0.5	0	5	5	100
1	0	5	5	100

项目 Item	对照组 Control group	TBHQ组 TBHQ group	PM_2.5组 PM_2.5 group	PM_2.5+ TBHQ组 PM_2.5+ TBHQ group	P值 P value
绝对重量Absolute weight (g)
种蛋重量 Egg weight	42.83 ± 1.33	41.53 ± 0.96	43.05 ± 1.08	40.41 ± 0.58	0.45
胚胎重量 Embryo weight	24.13 ± 0.75	22.52 ± 0.81	23.66 ± 1.19	23.47 ± 0.37	0.44
肺组织重量 Lung tissue weight	0.18 ± 0.02	0.17 ± 0.01	0.20 ± 0.02	0.17 ± 0.01	0.14
相对重量Relative weight
胚蛋比 The ratio of embryo and egg (g·g^-1)	0.57 ± 0.02	0.54 ± 0.02	0.55 ± 0.03	0.58 ± 0.01	0.94
肺胚比 The ratio of lung and embryo (mg·g^-1)	0.73 ± 0.09	0.75 ± 0.04	0.84 ± 0.08	0.74 ± 0.05	0.95

基因 Gene	对照组 Control group	TBHQ组 TBHQ group	PM_2.5组 PM_2.5 group	PM_2.5 + TBHQ组 PM_2.5 and TBHQ group
NLRP3	1.00 ± 0.19ab	0.62 ± 0.09 b	1.29 ± 0.13 a	1.56 ± 0.18a
Caspase-1	1.00 ± 0.10 c	1.63 ± 0.10 b	1.35 ± 0.08 bc	2.44 ± 0.16a
IL-18	1.00 ± 0.13 ab	0.72 ± 0.14 bc	1.14 ± 0.03a	0.57 ± 0.0a5c
IL-1β	1.00 ± 0.12b	0.57 ± 0.12bc	1.62 ± 0.14a	0.37 ± 0.03c
RIPK1	1.00 ± 0.17b	0.75 ± 0.06b	1.66 ± 0.09a	1.07 ± 0.14b
RIPK3	1.00 ± 0.07b	1.44 ± 0.21 ab	1.20 ± 0.23b	1.92 ± 0.12a
MLKL	1.00 ± 0.30ab	0.37 ± 0.10b	1.74 ± 0.26a	0.42 ± 0.08b

Effects of TBHQ on Pyroptosis, Necroptosis and Inflammatory Damage of Chicken Embryonic Lung Tissues Induced by PM_2.5 from Chicken Houses

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Effects of TBHQ on Pyroptosis, Necroptosis and Inflammatory Damage of Chicken Embryonic Lung Tissues Induced by PM2.5 from Chicken Houses

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Effects of TBHQ on Pyroptosis, Necroptosis and Inflammatory Damage of Chicken Embryonic Lung Tissues Induced by PM_2.5 from Chicken Houses