杀螟硫磷暴露对大鼠肝细胞的代谢毒性及机制研究

doi:10.19586/j.2095-2341.2024.0005

生物技术进展 ›› 2024, Vol. 14 ›› Issue (3): 459-465.DOI: 10.19586/j.2095-2341.2024.0005

• 研究论文 • 上一篇

杀螟硫磷暴露对大鼠肝细胞的代谢毒性及机制研究

李一凡¹(), 冯伟伟¹, 茆广华¹, 赵婷², 陈瑶¹, 罗梦娜¹, 耿鑫¹, 吴朝琼¹, 仰榴青², 吴向阳¹()

^1.江苏大学环境与安全工程学院，江苏镇江 212013
^2.江苏大学化学化工学院，江苏镇江 212013

收稿日期:2023-11-09 接受日期:2024-03-05 出版日期:2024-05-25 发布日期:2024-06-18
通讯作者: 吴向阳
作者简介:李一凡 E-mail：616513065@qq.com；
基金资助:
国家自然科学基金面上项目(22276079)

Metabolic Toxicity and Mechanism of Fenitrothion Exposure on BRL Cells

Yifan LI¹(), Weiwei FENG¹, Guanghua MAO¹, Ting ZHAO², Yao CHEN¹, Mengna LUO¹, Xin GENG¹, Chaoqiong WU¹, Liuqing YANG², Xiangyang WU¹()

^1.School of the Environment and Safety Engineering，Jiangsu University，Jiangsu Zhenjiang 212013，China
^2.School of Chemistry and Chemical Engineering，Jiangsu University，Jiangsu Zhenjiang 212013，China

Received:2023-11-09 Accepted:2024-03-05 Online:2024-05-25 Published:2024-06-18
Contact: Xiangyang WU

摘要/Abstract

摘要：

为了研究低剂量杀螟硫磷(fenitrothion-O-analog, FNT)暴露对大鼠肝细胞(buffalo rat liver cells,BRL)的代谢毒性，并通过作用于体外的一系列指标分析其作用机制，分别对空白对照组和杀螟硫磷暴露组(13.78、27.55、55.10 μg·mL^-1)暴露48 h，观察其对BRL内糖代谢、胰岛素敏感和糖原合成信号通路中蛋白表达的影响。实验结果表明，杀螟硫磷暴露能够显著抑制BRL细胞的活力，半数抑制浓度（IC₅₀）为275.5 μg·mL^-1。杀螟硫磷暴露使超氧化物歧化酶（superoxide dismutase，SOD）和乙酰胆碱酯酶AchE活力显著降低(P<0.01)，其体内丙二醛（malonaldehyde，MDA）含量显著增加(P<0.01)，引起细胞氧化损伤。杀螟硫磷暴露使其细胞内糖原、胰岛素和葡萄糖激酶含量显著降低(P<0.01)，增加胰岛素抵抗。杀螟硫磷暴露显著下调胰岛素敏感信号通路中IRS的表达，抑制IRS的磷酸化，并显著上调AKT和PI3K的表达，显著上调糖原合成信号通路中GSK-3α和GSK-3β的表达。因此，杀螟硫磷暴露使BRL产生糖代谢毒性的机制是通过氧化应激诱发胰岛素抵抗，从而改变糖代谢相关信号通路中蛋白的表达而实现的。

关键词: 杀螟硫磷, 糖代谢, 代谢毒性, 信号通路

Abstract:

To study the metabolic toxicity of low dose fenitrothion （FNT） exposure on rat BRL cells， and analyze its mechanism through a series of indicators in vitro， the control group and fenitrothion exposure groups（13.78， 27.55 and 55.10 μg·mL^-1） were exposed for 48 h respectively. We observed its effects on the intracellular glucose metabolism level in BRL cells， the expression of insulin sensitivity related proteins and glycogen synthesis related proteins in the signaling pathway of glucose metabolism. The experiment results showed that the viability of BRL cells was significantly inhibited by fenitrothion exposure with an IC₅₀ of 275.5 μg·mL^-1. Exposure to fenitrothion significantly decreased SOD and AchE activities （P<0.01）， increased the content of MDA （P<0.01）， and caused oxidative damage to cells. Exposure to fenitrothion significantly reduced intracellular glycogen content， insulin concentration， and glucokinase content （P<0.01）， increasing insulin resistance. Fenitrothion exposure significantly reduced IRS content， inhibited IRS phosphorylation， and significantly increased AKT， PI3K， GSK-3α and GSK-3β content. Therefore， The mechanism of glucose metabolism toxicity of fenitrothion exposure on BRL cells is that oxidative stress induces insulin resistance and changes the expression of proteins in the signaling pathways related to glucose metabolism.

Key words: fenitrothion, glucose metabolism, metabolic toxicity, signaling pathways

中图分类号:

Q493.4

李一凡, 冯伟伟, 茆广华, 赵婷, 陈瑶, 罗梦娜, 耿鑫, 吴朝琼, 仰榴青, 吴向阳. 杀螟硫磷暴露对大鼠肝细胞的代谢毒性及机制研究[J]. 生物技术进展, 2024, 14(3): 459-465.

Yifan LI, Weiwei FENG, Guanghua MAO, Ting ZHAO, Yao CHEN, Mengna LUO, Xin GENG, Chaoqiong WU, Liuqing YANG, Xiangyang WU. Metabolic Toxicity and Mechanism of Fenitrothion Exposure on BRL Cells[J]. Current Biotechnology, 2024, 14(3): 459-465.

图/表 8

图1 MTT法检测不同浓度杀螟硫磷对BRL细胞活力的影响注：*和**分别表示与空白对照组相比较在P<0.05和P<0.01水平有统计学差异。

Fig. 1 Effects of different concentrations of fenitrothion on the viability of BRL cells by MTT assay

图2 杀螟硫磷暴露对氧化应激相关酶的影响注：**表示与空白对照组相比较在P<0.01水平有统计学差异。

Fig. 2 Effects of fenitrothion exposure on oxidative stress-related enzymes

图3 杀螟硫磷暴露对AchE活性的影响注：*和**分别表示与空白对照组相比较在P<0.05和P<0.01水平有统计学差异。

Fig. 3 Effects of fenitrothion exposure on acetylcholinesterase activity

图4 杀螟硫磷暴露对糖原含量的影响注：**表示与空白对照组相比较在P<0.01水平有统计学差异。

Fig. 4 Effects of fenitrothion exposure on glycogen content

图5 杀螟硫磷暴露对胰岛素含量的影响注：*和**分别表示与空白对照组相比较在P<0.05和P<0.01水平有统计学差异。

Fig. 5 Effects of fenitrothion exposure on insulin content

图6 杀螟硫磷暴露对糖代谢相关酶的影响注：**表示与空白对照组相比较在P<0.01水平有统计学差异。

Fig. 6 Effects of fenitrothion exposure on glucose metabolism related enzymes

图7 杀螟硫磷对BRL细胞胰岛素敏感信号通路中相关蛋白表达的影响A：杀螟硫磷对BRL细胞胰岛素敏感信号通路相关蛋白表达影响的电泳结果；B：杀螟硫磷对BRL细胞ATK表达的影响；C：杀螟硫磷对BRL细胞p-ATK表达的影响；D：杀螟硫磷对BRL细胞IRS表达的影响；E：杀螟硫磷对BRL细胞p-IRS表达的影响；F：杀螟硫磷对BRL细胞PI3K表达的影响。*和**分别表示与空白对照组相比较在P<0.05和P<0.01水平有统计学差异。

Fig. 7 Effects of fenitrothion on the expression of related proteins in insulin sensitive signaling pathway of BRL cells

图8 杀螟硫磷对BRL细胞糖原合成信号通路中相关蛋白表达的影响A：杀螟硫磷对BRL细胞糖原合成信号通路中相关蛋白表达影响的电泳结果；B：杀螟硫磷对BRL细胞GSK-3α表达的影响；C：杀螟硫磷对BRL细胞GSK-3β表达的影响。**表示与空白对照组相比较在P<0.01水平有统计学差异。

Fig. 8 Effects of fenitrothion on the expression of related proteins in glycogen synthesis signaling pathway of BRL cells

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杀螟硫磷暴露对大鼠肝细胞的代谢毒性及机制研究

Metabolic Toxicity and Mechanism of Fenitrothion Exposure on BRL Cells

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