Stereoselective effects and its potential mechanism of ofloxacin on the growth and development of Rana nigromaculata: mainly liver lipid metabolism

Abstract

Most research on antibiotics in the environment disregards chiral antibiotics, such as ofloxacin (OF). In this study, tadpoles of Rana nigromaculata were exposed to 1 μg/L OF and levofloxacin (LVFX, an enantiomer of OF) for 75 d. Compared with dextrofloxacin, LVFX treatment had a greater effect on the inhibition of bodyweight, body length, development stage, and pathological liver damage. Therefore, OF exerts a stereoselective inhibitory effect on both growth and development, which is consistent with the results at the systemic metabolism level. Transcriptomic analysis revealed that the differentially expressed genes between OF and LVFX were mainly immune related. Targeted metabolomics showed that the stereoselective biological effect of OF on R. nigromaculata was caused by differences in contents of PE-O 16:0–22:4, PE 16:0–14:0, TAG 45:0–FA16:0, PE-P 18:0–16:0, PE 16:0–16:0, PC 16:0–22:4 + AcO, PE 18:1–18:3, PC 16:1–18:1 + AcO, and PC 18:1–18:3 + AcO. Furthermore, two enantiomers of OF were selectively bound to enzymes related to lipid metabolism. This study provides both theoretical and practical references for the accurate evaluation and scientific control of the ecological risk of chiral antibiotics.

Graphical abstract

Keywords

Chiral antibiotics / Amphibians / Targeted metabolomics / Molecular docking / Obesity

Highlight

	● Ofloxacin has stereoselective inhibitory effect on tadpole growth and development.
	● Inhibition effect of ofloxacin on tadpole development can last for 75 d.
	● Differential genes between ofloxacin and its enantiomer are mainly immune-related.
	● Ofloxacin and its enantiomer show the potential to promote obesity.
	● Two enantiomers were selectively bound to enzymes related to lipid metabolism.

References

[1] Althaqafi A, Ali M, Alzahrani Y, Ming L C, Hussain Z. (2021). How safe are fluoroquinolones for diabeticpatients? A systematic review of dysglycemic and neuropathic effects of fluoroquinolones. Therapeutics and Clinical Risk Management, 17: 1083–1090

[2] Arenas M, Martín J, Santos J L, Aparicio I, Alonso E. (2022). Enantioselective behavior of environmental chiral pollutants: a comprehensive review. Critical Reviews in Environmental Science and Technology, 52(17): 2995–3034

[3] Blanc M, Alfonso S, Begout M L, Barrachina C, Hyotylainen T, Keiter S H, Cousin X. (2021). An environmentally relevant mixture of polychlorinated biphenyls (PCBs) and polybrominated diphenylethers (PBDEs) disrupts mitochondrial function, lipid metabolism and neurotransmission in the brain of exposed zebrafish and their unexposed F2 offspring. Science of the Total Environment, 754: 142097

[4] Chen H, Chen Z, Hu L, Tang F, Kuang D, Han J, Wang Y, Zhang X, Cheng Y, Meng J. . (2024). Application of wastewater-based epidemiological monitoring of COVID-19 for disease surveillance in the city. Frontiers of Environmental Science & Engineering, 18(8): 98

[5] Chen Z, Han S, Zheng P, Zhang J, Zhou S, Jia G. (2022). Landscape of lipidomic metabolites in gut-liver axis of Sprague-Dawley rats after oral exposure to titanium dioxide nanoparticles. Particle and Fibre Toxicology, 19(1): 53

[6] Cristóvão M B, Tela S, Silva A F, Oliveira M, Bento-Silva A, Bronze M R, Crespo M T B, Crespo J G, Nunes M, Pereira V J. (2020). Occurrence of antibiotics, antibiotic resistance genes and viral genomes in wastewater effluents and their treatment by a pilot scale nanofiltration unit. Membranes, 11(1): 9

[7] Du J, Liu Q, Pan Y, Xu S, Li H, Tang J. (2023). The research status, potential hazards and toxicological mechanisms of fluoroquinolone antibiotics in the environment. Antibiotics, 12(6): 1058

[8] Feng M, Wang Z, Dionysiou D D, Sharma V K. (2018). Metal-mediated oxidation of fluoroquinolone antibiotics in water: A review on kinetics, transformation products, and toxicity assessment. Journal of Hazardous Materials, 344: 1136–1154

[9] Ghosh G, Hanamoto S, Yamashita N, Huang X, Tanaka H. (2016). Antibiotics removal in biological sewage treatment plants. Pollution, 2(2): 131–139

[10] Gosner K L J H. (1960). A simplified table for staging anuran embryos and larvae with notes on identification. Herpetologica, 16(3): 183–90

[11] Grabner G F, Xie H, Schweiger M, Zechner R. (2021). Lipolysis: cellular mechanisms for lipid mobilization from fat stores. Nature Metabolism, 3(11): 1445–1465

[12] Guo W, Yang Y, Zhou X, Ming R, Hu D, Lu P. (2022). Insight into the toxic effects, bioconcentration and oxidative stress of acetamiprid on Rana nigromaculata tadpoles. Chemosphere, 305: 135380

[13] Hamid N, Junaid M, Wang Y, Pu S Y, Jia P P, Pei D S. (2021). Chronic exposure to PPCPs mixture at environmentally relevant concentrations (ERCs) altered carbohydrate and lipid metabolism through gut and liver toxicity in zebrafish. Environmental Pollution, 273: 116494

[14] Han Y, Ma Y, Yao S, Zhang J, Hu C. (2021). In vivo and in silico evaluations of survival and cardiac developmental toxicity of quinolone antibiotics in zebrafish embryos (Danio rerio). Environmental Pollution, 277: 116779

[15] Hao Z M, Ye Y F, Zhang Y K, Yang S F, Ye X L. (2016). Lipoprotein lipase and lipid profiles in plasma and placenta from normal pregnancies compared with patients with intrahepatic cholestasis of pregnancy. European Journal of Obstetrics, Gynecology, and Reproductive Biology, 203: 279–285

[16] Hennig B, Ormsbee L, Mcclain C J, Watkins B A, Blumberg B, Bachas L G, Sanderson W, Thompson C, Suk W A. (2012). Nutrition can modulate the toxicity of environmental pollutants: implications in risk assessment and human health. Environmental Health Perspectives, 120(6): 771–774

[17] Hu Y, Hu J, Li W, Gao Y, Tian Y. (2021). Changes of embryonic development, locomotor activity, and metabolomics in zebrafish co-exposed to chlorpyrifos and deltamethrin. Journal of Applied Toxicology, 41(9): 1345–1356

[18] Jantsch J, Binger K J, Müller D N, Titze J. (2014). Macrophages in homeostatic immune function. Frontiers in Physiology, 5: 146

[19] Ji Y, Sun S, Xu A, Bhargava P, Yang L, Lam K S, Gao B, Lee C H, Kersten S, Qi L. (2012). Activation of natural killer T cells promotes M2 Macrophage polarization in adipose tissue and improves systemic glucose tolerance via interleukin-4 (IL-4)/STAT6 protein signaling axis in obesity. Journal of Biological Chemistry, 287(17): 13561–13571

[20] Juhl C R, Burgdorf J, Knudsen C, Lubberding A F, Veedfald S, Isaksen J L, Hartmann B, Frikke-Schmidt R, Mandrup-Poulsen T, Holst J J. . (2023). A randomized, double-blind, crossover study of the effect of the fluoroquinolone moxifloxacin on glucose levels and insulin sensitivity in young men and women. Diabetes, Obesity & Metabolism, 25(1): 98–109

[21] Kan X, Yu X, Zhao W, Lyu S, Sun S, Yu G, Sui Q. (2023). Screening of indicator pharmaceuticals and personal care products in landfill leachates: a case study in Shanghai, China. Frontiers of Environmental Science & Engineering, 17(9): 116

[22] Kaziem A E, Gao B, Li L, Zhang Z, He Z, Wen Y, Wang M H. (2020). Enantioselective bioactivity, toxicity, and degradation in different environmental mediums of chiral fungicide epoxiconazole. Journal of Hazardous Materials, 386: 121951

[23] Kergaravat S V, Hernandez S R, Maria Gagneten A. (2021). Second-, third- and fourth-generation quinolones: ecotoxicity effects on Daphnia and Ceriodaphnia species. Chemosphere, 262: 127823

[24] Khetarpal S A, Vitali C, Levin M G, Klarin D, Park J, Pampana A, Millar J S, Kuwano T, Sugasini D, Subbaiah P V. . (2021). Endothelial lipase mediates efficient lipolysis of triglyceride-rich lipoproteins. PLOS Genetics, 17(9): e1009802

[25] Kobayashi J, Miyashita K, Nakajima K, Mabuchi H. (2015). Hepatic lipase: a comprehensive view of its role on plasma lipid and lipoprotein metabolism. Journal of Atherosclerosis and Thrombosis, 22(10): 1001–1011

[26] KoÇak E, Özkul KoÇak C J J. (2020). Metabolic response of Escherichia coli to subinhibitory concentration of ofloxacin. Journal of Research in Pharmacy, 24(4): 593–601

[27] Kotas M E, Lee H Y, Gillum M P, Annicelli C, Guigni B A, Shulman G I, Medzhitov R. (2011). Impact of CD1d deficiency on metabolism. PLoS One, 6(9): e25478

[28] Kou H, Ya J, Gao X, Zhao H. (2020). The effects of chronic lead exposure on the liver of female Japanese quail (Coturnix japonica): histopathological damages, oxidative stress and AMP-activated protein kinase based lipid metabolism disorder. Ecotoxicology and Environmental Safety, 190: 110055

[29] LaPensee C R, Lin G, Dent A L, Schwartz J. (2014). Deficiency of the transcriptional repressor B Cell Lymphoma 6 (Bcl6) is accompanied by dysregulated lipid metabolism. PLoS One, 9(6): e97090

[30] Legrand P, Rioux V J L. (2010). The complex and important cellular and metabolic functions of saturated fatty acids. Lipids, 45(10): 941–946

[31] Li W, Zhang S, Wang X, Yu J, Li Z, Lin W, Lin X. (2018). Systematically integrated metabonomic-proteomic studies of Escherichia coli under ciprofloxacin stress. Journal of Proteomics, 179: 61–70

[32] Lin H, Liu Z, Yang H, Lu L, C hen R, Zhang X, Zhong Y, Zhang H. (2022). Per- and polyfluoroalkyl substances (PFASs) impair lipid metabolism in Rana nigromaculata: a field investigation and laboratory study. Environmental Science & Technology, 56(18): 13222–13232

[33] Lips K R, Diffendorfer J, Mendelson J R, Sears M W. (2008). Riding the wave: Reconciling the roles of disease and climate change in amphibian declines. PLoS Biology, 6(3): 441–454

[34] Luedtke J A, Chanson J, Neam K, Hobin L, Maciel A O, Catenazzi A, Borzee A, Hamidy A, Aowphol A, Jean A. . (2024). Author Correction: Ongoing declines for the world’s amphibians in the face of emerging threats. Nature, 625(7993): E2

[35] Parsons J B, Rock C O J C O I M (2011). Is bacterial fatty acid synthesis a valid target for antibacterial drug discovery? Current Opinion in Microbiology, 14(5): 544–549

[36] Petersen M C, Vatner D F, Shulman G I. (2017). Regulation of hepatic glucose metabolism in health and disease. Nature Reviews. Endocrinology, 13(10): 572–587

[37] Petrie B, Camacho-Munoz D. (2021). Analysis, fate and toxicity of chiral non-steroidal anti-inflammatory drugs in wastewaters and the environment: a review. Environmental Chemistry Letters, 19(1): 43–75

[38] Ren Z, Xu H, Wang Y, Li Y, Han S, Ren J. (2021). Combined toxicity characteristics and regulation of residual quinolone antibiotics in water environment. Chemosphere, 263: 128301

[39] Riaz L, Mahmood T, Kamal A, Shafqat M, Rashid A. (2017). Industrial release of fluoroquinolones (FQs) in the waste water bodies with their associated ecological risk in Pakistan. Environmental Toxicology and Pharmacology, 52: 14–20

[40] Ribeiro A R L, Maia A S, Ribeiro C, Tiritan M E. (2020). Analysis of chiral drugs in environmental matrices: Current knowledge and trends in environmental, biodegradation and forensic fields. Trends in Analytical Chemistry, 124: 115783

[41] Ruderman N, Flier J S. (2001). Cell biology-Chewing the fat-ACC and energy balance. Science, 291(5513): 2558–2559

[42] Sang Y, Zhang J, Liu K, Wang Q, Wang S, Sheng J, Wang L, Zhang D, Li X, Cao H. . (2021). Antibiotics biomonitored in urine and obesogenic risk in a community-dwelling elderly population. Ecotoxicology and Environmental Safety, 210: 111863

[43] Sanganyado E, Lu Z, Fu Q, Schlenk D, Gan J. (2017). Chiral pharmaceuticals: a review on their environmental occurrence and fate processes. Water Research, 124: 527–542

[44] Stuart S N, Chanson J S, Cox N A, Young B E, Rodrigues A S L, Fischman D L, Waller R W. (2004). Status and trends of amphibian declines and extinctions worldwide. Science, 306(5702): 1783–1786

[45] Wang B, Tontonoz P. (2018). Liver X receptors in lipid signalling and membrane homeostasis. Nature Reviews. Endocrinology, 14(8): 452–463

[46] Wang R, Tang R, Li B, Ma X, Schnabl B, Tilg H. (2021). Gut microbiome, liver immunology, and liver diseases. Cellular & Molecular Immunology, 18(1): 4–17

[47] Wright H T, Reynolds K A. (2007). Antibacterial targets in fatty acid biosynthesis. Current Opinion in Microbiology, 10(5): 447–453

[48] Xu H, Jia Y, Sun Z, Su J, Liu Q S, Zhou Q, Jiang G. (2022). Environmental pollution, a hidden culprit for health issues. Eco-Environment & Health, 1(1): 31–45

[49] Yao Z, Lin Z, Wang T, Tian D, Zou X, Gao Y, Yin D J. (2013). Using molecular docking-based binding energy to predict toxicity of binary mixture with different binding sites. Chemosphere, 92(9): 1169–1176

[50] Yu X H, Cai Y, Keereetaweep J, Wei K, Chai J, Deng E, Liu H, Shanklin J. (2021). Biotin attachment domain-containing proteins mediate hydroxy fatty acid-dependent inhibition of acetyl CoA carboxylase. Plant Physiology, 185(3): 892–901

[51] Yuan J, Huang X, Gu J, Yuan Y, Liu Z, Zou H, Bian J. (2023). Honokiol reduces cadmium-induced oxidative injury and endosomal/lysosomal vacuolation via protecting mitochondrial function in quail (Coturnix japonica) liver tissues. Science of the Total Environment, 857: 159626

[52] Zhang L, Shen L, Qin S, Cui J, Liu Y. (2020). Quinolones antibiotics in the Baiyangdian Lake, China: occurrence, distribution, predicted no-effect concentrations (PNECs) and ecological risks by three methods. Environmental Pollution, 256: 113458

[53] Zhang W, Teng M, Zhao L, Chen L. (2023a). Study effect and mechanism of ofloxacin and levofloxacin on development of Rana nigromaculata tadpoles based on the hypothalamus-pituitary-thyroid axis. Ecotoxicology and Environmental Safety, 259: 114985

[54] Zhang X, Razanajatovo M R, Du X, Wang S, Feng L, Wan S, Chen N, Zhang Q. (2023b). Well-designed protein amyloid nanofibrils composites as versatile and sustainable materials for aquatic environment remediation: a review. Eco-Environment & Health, 2(4): 264–277

[55] Zhang Y, Sun W, Wang B, Liu Z, Liu Z, Zhang X, Wang B, Han Y, Zhang H. (2024). Metabolomics reveals the lipid metabolism disorder in Pelophylax nigromaculatus exposed to environmentally relevant levels of microcystin-LR. Environmental Pollution, 358: 124458

[56] Zhou Z, Zhang Z, Feng L, Zhang J, Li Y, Lu T, Qian H. (2020). Adverse effects of LEVO floxacin and oxytetracycline on aquatic microbial communities. Science of the Total Environment, 734: 139499

[57] Zhu L, Santiago-Schuebel B, Xiao H, Hollert H, Kueppers S. (2016). Electrochemical oxidation of fluoroquinolone antibiotics: mechanism, residual antibacterial activity and toxicity change. Water Research, 102: 52–62

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