Difenoconazole disrupts carp intestinal physical barrier and causes inflammatory response via triggering oxidative stress and apoptosis

Xinyu Wu; Hairui Han; Kunmei Xie; Nana He; Zuwang Yang; Xiaohui Jin; Shaojie Ma; Jingquan Dong

doi:10.1016/j.pestbp.2023.105507

Difenoconazole disrupts carp intestinal physical barrier and causes inflammatory response via triggering oxidative stress and apoptosis

Pestic Biochem Physiol. 2023 Aug:194:105507. doi: 10.1016/j.pestbp.2023.105507. Epub 2023 Jun 24.

Authors

Xinyu Wu¹, Hairui Han¹, Kunmei Xie¹, Nana He¹, Zuwang Yang¹, Xiaohui Jin¹, Shaojie Ma², Jingquan Dong³

Affiliations

¹ Jiangsu Key Laboratory of Marine Bioresources and Environment, Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005, China.
² Jiangsu Key Laboratory of Marine Bioresources and Environment, Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005, China. Electronic address: mashaojie@jou.edu.cn.
³ Jiangsu Key Laboratory of Marine Bioresources and Environment, Co-Innovation Center of Jiangsu Marine Bio-industry Technology, Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening, Jiangsu Ocean University, Lianyungang 222005, China. Electronic address: 2018000029@jou.edu.cn.

PMID: 37532360
DOI: 10.1016/j.pestbp.2023.105507

Abstract

As a common fungicide, difenoconazole (DFZ) is widespread in the natural environment and poses many potential threats. Carp makes up a significant proportion of China's freshwater aquaculture population and are vulnerable to the DFZ. Therefore, this study investigated the effects of DFZ (0.488 mg/L and 1.953 mg/L) exposure for 4 d on the intestinal tissues of carp and explored the mechanisms. Specifically, DFZ exposure caused pathological damage to the intestinal tissues of carp, reducing the expression levels of intestinal tight junction proteins, and leading to damage to the intestinal barrier. In addition, DFZ exposure activated the NF-κB signaling pathway, increasing the levels of pro-inflammatory factors (TNF-α, IL-1β, IL-6) and decreasing the levels of anti-inflammatory factors (IL-10, TGF-β1). As disruption of the intestinal barrier is closely linked to oxidative stress and apoptosis, we have conducted research in both areas for this reason. The results showed that DFZ exposure elevated reactive oxygen species in carp intestines, decreased antioxidant enzyme activity, and suppressed the expression of oxidative stress-related genes. TUNEL results showed that DFZ induced the onset of apoptosis. In addition, the expression levels of apoptosis-related genes and proteins were examined. Western blotting results showed that DFZ could upregulate the protein expression levels of Bax, Cytochrome C and downregulate the protein levels of Bcl-2. qPCR results showed that DFZ could upregulate the transcript levels of Bax, Caspase-3, Caspase-8 and Caspase-9 and downregulate the transcript levels of Bcl-2 transcript levels. This suggests that DFZ can induce apoptosis of mitochondrial pathway in carp intestine. In conclusion, DFZ can induce oxidative stress and apoptosis in carp intestine, leading to the destruction of intestinal physical barrier and the occurrence of inflammation. Our data support the idea that oxidative stress and apoptosis are important triggers of pesticide-induced inflammatory bowel illness.

Keywords: Apoptosis; Difenoconazole; Inflammatory response; Intestinal physical barrier; Oxidative stress.

MeSH terms

Animals
Antioxidants / pharmacology
Apoptosis
Carps* / metabolism
Fish Proteins / genetics
Fish Proteins / metabolism
Fish Proteins / pharmacology
Intestines
NF-kappa B / metabolism
Oxidative Stress
bcl-2-Associated X Protein / metabolism

Substances

difenoconazole
bcl-2-Associated X Protein
Fish Proteins
Antioxidants
NF-kappa B