| N-乙酰半胱氨酸对双酚A诱导的猪肾细胞凋亡和炎症反应的抑制作用 |
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| 引用本文： | 陶文静,张子婷,刘媛,宋丹,李向臣. N-乙酰半胱氨酸对双酚A诱导的猪肾细胞凋亡和炎症反应的抑制作用[J]. 中国农业科学, 2023, 56(3): 549-558. DOI: 10.3864/j.issn.0578-1752.2023.03.012 |
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| 作者姓名： | 陶文静 张子婷 刘媛 宋丹 李向臣 |
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| 作者单位： | 浙江农林大学动物科技学院/动物医学院/浙江省畜禽绿色生态健康养殖应用技术研究重点实验室，杭州 311300 |
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| 基金项目： | 浙江省自然科学基金(LQ22C170001);浙江农林大学科研发展基金(22020FR080);国家科技基础条件平台子课题(TZDWZYK2019-08) |
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| 摘 要： | 【背景】双酚A(BPA)广泛应用于塑料材料工业制造，其从塑料制品中渗出，暴露在食物、水、土壤和空气等多种环境介质中，导致动物长期接触，并经过胎盘和母乳传递给后代，干扰动物生长和发育，对动物生长性能和生产效率产生不利影响。N-乙酰半胱氨酸（NAC）作为公认的强效抗氧化剂，能够调节氧化应激、细胞凋亡、炎症等多种病理生理过程。然而，NAC对BPA诱导的猪肾细胞损伤的调节作用尚不清楚。【目的】探究抗氧化剂NAC在BPA诱导的PK15细胞凋亡和炎症反应中的潜在作用，为NAC在对抗BPA诱导的猪肾细胞损伤中的应用研究提供科学依据。【方法】选取PK15细胞为试验材料，采用相应的抗氧化检测试剂盒分别测定细胞内过氧化氢酶（CAT）、总超氧化物歧化酶（T-SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性变化；设置不同浓度NAC(0、2、5和10 mmol·L-1)预处理，并与BPA共处理PK15细胞，CCK-8检测细胞活力以筛选最佳的NAC作用浓度；实时荧光定量PCR(qRT-PCR)检测凋亡相关基因（BAX、BCL-2和Caspase3）表达和炎症相关基因（IL-8、IL-6、IL-1β和TNF-α）m...
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| 关 键 词： | 双酚A N-乙酰半胱氨酸 细胞凋亡 炎症反应 PK15细胞 氧化应激 |
| 收稿时间： | 2021-11-11 |
Inhibitory Effect of N-acetylcysteine on Bisphenol A-Induced Apoptosis and Inflammatory Response in Porcine Kidney Cells |
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| TAO WenJing,ZHANG ZiTing,LIU Yuan,SONG Dan,LI XiangChen. Inhibitory Effect of N-acetylcysteine on Bisphenol A-Induced Apoptosis and Inflammatory Response in Porcine Kidney Cells[J]. Scientia Agricultura Sinica, 2023, 56(3): 549-558. DOI: 10.3864/j.issn.0578-1752.2023.03.012 |
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| Authors: | TAO WenJing ZHANG ZiTing LIU Yuan SONG Dan LI XiangChen |
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| Affiliation: | College of Animal Science and Technology/College of Veterinary Medicine, Zhejiang A&F University/Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Hangzhou 311300 |
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| Abstract: | 【Background】 Bisphenol A (BPA) is widely used in the industrial manufacturing of plastic materials, which seeps out from plastic products and exposes to various environmental media such as food, water, soil, and air, resulting in long-term exposure of animals. It is passed to offspring through the placenta and breast milk, interfering with animal growth and development and adversely affecting animal growth performance and production efficiency. N-acetylcysteine (NAC), as a recognized potent antioxidant, can regulate various pathophysiological processes, such as oxidative stress, apoptosis, and inflammation. However, the regulatory role of NAC on BPA-induced porcine kidney cell injury remains unclear. 【Objective】This study aimed to explore the potential role of the antioxidant NAC on BPA-induced apoptosis and inflammatory responses in PK15 cells.【Method】 PK15 cells were selected as experimental materials, and the activities of catalase (CAT), total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) were measured by the corresponding antioxidant detection kits. PK15 cells were treated with different concentrations of NAC (0, 2, 5, 10 mmol·L-1) and then co-treated with BPA, and then the cell viability was detected by CCK-8 to select the optimal concentration of NAC. The expression of apoptosis-related genes (BAX, BCL-2 and Caspase3) and inflammatory genes (IL-8, IL-6, IL-1β and TNF-α) as well as protein expression were detected by real-time quantitative PCR (qRT-PCR) and western blotting. The number of apoptotic cells and nuclear factor kappa B (NF-κB) nuclear translocation were detected by immunofluorescence staining. 【Result】 The results showed that BPA significantly reduced the activities of CAT, T-SOD and GSH-Px in PK15 cells, compared with the control group (P<0.05). CCK-8 results showed that BPA significantly decreased PK15 cell viability in contrast to control group, whereas the different concentrations of NAC significantly promoted cell viability, and 5, 10 mmol·L-1 NAC pretreatment significantly promoted cell viability when compared with BPA alone. qRT-PCR and western blotting showed that BPA treatment significantly increased BAX and Caspase3 mRNA and protein expression, and decreased the BCL-2 mRNA and protein expression, whereas NAC pretreatment could reduce the increased BAX and Caspase3 expression and increase the decreased BCL-2 expression induced by BPA. Hoechst33258 fluorescence staining indicated that the cells treated with BPA showed strong blue fluorescence staining and obvious nuclear shrinkage, whereas NAC pretreated cell showed weak blue fluorescence. BPA treatment significantly increased the relative mRNA expression of inflammation-related factors (IL-8 and IL-6) (P<0.05), whereas NAC pretreatment inhibited BPA-induced increased inflammation-related factors (IL-8, IL-6 and IL-1β mRNA) relative expression. Immunofluorescence analysis of NF-κB nuclear translocation showed that NF-κB was mainly distributed in cytoplasm in the control group, whereas NF-κB was mainly distributed in the nucleus after BPA treatment and NAC pretreatment reduced nuclear NF-κB expression. 【Conclusion】 NAC significantly increased PK15 cell viability and inhibited PK15 cell apoptosis and inflammatory response induced by BPA. |
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| Keywords: | Bisphenol A N-acetylcysteine apoptosis inflammatory response PK15 cell oxidative stress |
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