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

【目的】探讨特丁基对苯二酚(tert-Butylhydroquinone,TBHQ)对鸡舍细颗粒物(fine particulate matter,PM_2.5)诱导鸡胚肺损伤的缓解作用及机制，为预防和缓解鸡舍PM_2.5污染引起的鸡呼吸道健康问题提供理论依据。【方法】选用14日龄鸡胚作为研究对象，首先建立鸡舍PM_2.5诱导鸡胚肺组织损伤模型，再利用鸡胚肺损伤模型研究TBHQ的缓解作用。在建立鸡舍PM_2.5诱导鸡胚肺组织损伤模型试验中，选取不同浓度的PM_2.5(0、0.25、0.5、1 mg·mL^-1)在14日龄鸡胚卵白处注射，5 d后，观察鸡胚存活率以及鸡胚肺组织形态，选取合适的PM_2.5处理浓度(0.25 mg·mL^-1)，建立鸡胚肺损伤模型。在TBHQ对PM_2.5诱导鸡胚肺损伤的影响试验中，将14日龄鸡胚分为对照组(生理盐水)、PM_2.5组(0.25 mg·mL

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

【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.