Introduction
Atrazine (ATR; 2‑chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) is a globally prevalent herbicide widely employed in agricultural systems to regulate broadleaf and gramineous weeds (Costa et al., 2020) due to its water solubility (Ying et al., 2005), ATR can lead to serious contamination of surface water and groundwater. Although ATR residues in maize grain are generally below the maximum residue limit (MRL, 0.01 mg/kg), a recent large-scale monitoring study of 358 maize kernel samples from three major maize production regions found that ATR residue level of some samples exceed the limit of the safety standard (Wei et al., 2026). In addition, a recent terminal residue and dietary risk assessment using HPLC-MS/MS showed that ATR residues in corn straw ranged from 0.05 to 0.17 mg/kg, indicating that ATR can still persist in crop (Cao et al., 2023).
Another study also has reported that ATR concentrations ranged from 0.69 to 19.59 mg/kg in soils, 7.85 to 23.85 mg/kg in grass, and 1.88 to 43.08 mg/kg in cereals, demonstrating ATR accumulation along the trophic chain (Irena et al., 2005). The above findings indicate that ATR can persist in crop residues and the environment even when grain residues comply with safety standards. Meanwhile, ATR tends to accumulate in ecological environment because of its high chemical stability and prolonged environmental persistence (Fakhouri et al., 2010).
Therefore, ATR can cause widespread environmental pollution and bioaccumulation by the food chain (Fang et al., 2018). The widespread application of ATR threatens public health and diverse organisms, including birds, aquatic life, and mammals (Remigio et al., 2024). ATR can cause dysfunction in multiple major organs, including the pancreas, testis, liver, muscle, and kidney (Hussain et al., 2011; Sagarkar et al., 2016; Zhang et al., 2018; Hadeed et al., 2022).
Therefore, clarifying the health risks of ATR to humans and animals is of enormous importance for public health and safety.
Kidney serves as the main excretory organ responsible for filtering blood and regulating body fluid composition (Levassort and Essig, 2024). Kidney serves a vital function in sustaining ionic and osmotic homeostasis (Danziger and Zeidel, 2015), which is essential for physiological functions such as nutrient reabsorption, waste excretion and acid-base balance (Foresto-Neto et al., 2024). Since the kidney is responsible for controlling ion and osmotic balance, it is a main target for environmental toxins.
A prospective study demonstrated that ATR application was associated with an increasing risk of renal cell carcinoma in humans (Andreotti et al., 2020). ATR exposure could cause renal functional impairment and oxidative stress-mediated renal dysfunction in Nile tilapia (Abdel-Warith et al., 2021). In mice, ATR exposure significantly induced renal injury, as evidenced by elevated serum levels of uric acid, and creatinine (Zou et al., 2025).
Similarly, ATR co-exposure with ethanol exacerbated renal injury by elevating serum uric acid, increasing malondialdehyde, and altering antioxidant enzymes in Wistar rats (Abarikwu et al., 2017). Taken together, kidney serves as a primary target of ATR toxicity. Ionic homeostasis is critical for maintaining a wide range of physiological processes, including transmembrane ionic balance, membrane potential, and pH regulation to maintain internal stability (Hoenig and Zeidel, 2014).
Ion imbalance of sodium ion (Na+), potassium ion (K+), calcium ion (Ca²⁺), and magnesium ion (Mg²⁺) can compromise essential renal processes, leading to impaired tubular function (McCormick and Ellison, 2015), reduced glomerular filtration, and ultimately resulting in renal dysfunction (Longden and Lederer, 2024). At the cellular level, ion imbalance can directly contribute to structural damage, including the loss of polarity in tubular epithelial cells and disruption of intercellular junctions (Tuncel et al., 2015), and can further activate apoptotic pathways (Wiegand et al., 2023). ATPases are major active transporters of cations that uphold steep concentration gradients across cell membranes (Morth et al., 2011), and are essential for maintenance of cellular and systemic water and ion homeostasis (Kumari and Rathore, 2020).
Researchers have demonstrated that ATR caused ionic imbalance in the quail liver and heart by disrupting ATPase activities (Lin et al., 2016a). In mice, ATR-induced disturbances of renal ion and ATPase activity have been documented. (Guo et al., 2021).
Collectively, ionic imbalance and ATPase dysfunction are major contributors to ATR renal toxicity.
Ionic balance is closely related to water homeostasis, and water transport across biological membranes is a critical determinant of ion distribution and concentration (Verbalis, 2003; Login and Nejsum, 2023). As a category of transmembrane channel proteins, AQPs mediate the bidirectional movement of water across cell membranes, which follows the direction of osmotic gradients (Agre, 2006). AQPs fall into three major categories based on their molecular structural features and functional patterns.
As the first category, orthodox AQPs contain seven isoforms (AQP0, AQP1, AQP2, AQP4, AQP5, AQP6, and AQP8), with a primary function of selectively conducting water across membranes. The second category, glycerol AQPs, comprises AQP3, AQP7, AQP9, and AQP10, which mediate the transport of water as well as small neutral molecules. The third group is super AQPs, which also referred to as unorthodox AQPs, and this group includes AQP11 and AQP12 (Soveral and Casini, 2017).
Additionally, AQP1, AQP2, AQP3, AQP4, AQP6, AQP7, and AQP11 are mainly expressed in renal tissue (Su et al., 2020). Studies have shown that environmental toxicants di-(2-ethylhexyl) phthalate (DEHP) can dysregulate AQPs expression, leading to renal injury and ion imbalance (Jiang et al., 2021; Li et al., 2023). Specifically, ATR has been demonstrated to impair renal AQPs expression, with AQP2 being particularly affected (Guo et al., 2021).
Thus, modulating ATPases activities and AQPs expression collectively governs renal water and ion balance, representing a potential strategy to alleviate ATR-induced nephrotoxicity.
Selenium (Se) is a well-known essential trace element with well-documented antioxidant, anti-inflammatory, and detoxifying capacities (Genchi et al., 2023). Selenium-enriched yeast (Yeast-Se), an organic form of Se, boasts enhanced bioavailability and a safer profile than its inorganic counterparts (Ferrari et al., 2023). Yeast-Se can mitigate heavy metal-induced ion dysregulation and organ damage in poultry (Wu et al., 2022).
Yeast-Se supplementation attenuated cadmium-caused liver injury and restored ionic balance in chickens (Wang et al., 2020). Furthermore, Yeast-Se has been reported to protect against necroptosis induced by cadmium in chicken kidney via miR-26a-5p/PTEN/PI3K/AKT signaling pathway (Chen et al., 2021). Additionally, Yeast-Se has been demonstrated to protect against aluminum-triggered renal kidney injury and ion dysregulation, further supporting its role in maintaining kidney health (Cao et al., 2018).
Yeast-Se was more readily absorbed and accumulated to significantly higher concentrations in the kidneys than inorganic selenium sources (Zhou et al., 2020), indicating considerable nephroprotective effects. It is worth noting that, Yeast-Se administration alleviated hepatic ion disturbances caused by ATR in quails. (Wang et al., 2024).
However, the nephroprotective role of Yeast-Se against ATR-triggered renal ion imbalance and its underlying mechanism remain to be fully understood.
Quail (Coturnix japonica) possesses unique biological characteristics that make it an important sentinel species for ecotoxicological monitoring and a reliable model for assessing ecological risks (Zhang et al., 2023). Quail serves as an ideal system for investigating ATR toxicity. Therefore, this present study aimed to investigate whether Yeast-Se protected against ATR-induced renal damage in quails by maintaining ATPases activity and AQPs expression.
The findings intended to shed new light on the nephroprotective mechanisms of Yeast-Se and support its application as a dietary supplement to mitigate environmental pollutants-induced renal toxicity in poultry.
Materials and methods
Animals and experimental protocol
In this study, a total of 80 one-day-old Japanese quails (Coturnix japonica) underwent a 7-day acclimation. After 7 days of acclimation, quails were divided into four groups at random (each containing 20 quails). The specific dietary regimen and ATR exposure scheme for each group are detailed in Table 1.
ATR exposure doses were based on avian LD50 and NOAEL of ATR (de la Casa-Resino et al., 2012; USEPA, 2002). 500 mg/kg ATR was equal to one tenth of LD50 and twice of NOAEL, which could reveal the adverse effects and the target organ and organelle of ATR’s toxicity. (Lin et al., 2016a; Zhang et al., 2024; Wang et al., 2024).
All quails were housed under identical conditions. The total experimental period was 28 days. Following the final gavage on day 28, all animals were euthanized, and serum and kidney samples were obtained to be used in the following experiments.
The Animal Care and Use Committee of Henan Agricultural University (Approval No. HNND2021031005) has formally reviewed and approved the animal study protocol, and all experimental operations strictly abide by the National Institutes of Health’s “Guide for the Care and Use of Laboratory Animals” (8th edition, 2010).Table 1Grouping and treatment protocols of experimental quails.GroupFull group nameDietary treatmentNumberConControl groupBasal diet20ATRATR exposure groupBasal diet + 500 mg/kg B.W. ATR by gavage20ATR + SeATR + Yeast-Se co-treatment groupBasal diet supplemented with 0.3 mg/kg Yeast-Se + 500 mg/kg B.W.
ATR by gavage20Yeast-SeYeast-Se groupBasal diet supplemented with 0.3 mg/kg Yeast-Se20
Tissue and histopathological evaluation
Renal tissues were immersed in 10% neutral formalin solution to complete fixation and sectioned serially at a thickness of 5 μm. Standard Hematoxylin and Eosin (H&E) staining was conducted on the tissue sections, which were subsequently subjected to microscopic examination (BA600, Digital Slice Browsing System, McAudi Sales Co., Ltd., Xiamen, China). The scan interface of the imaging system was applied to conduct the full pathological evaluation process.
For histomorphometric analysis, renal sections were evaluated by two independent pathologists. Tubular and glomerular lesions were respectively assigned semi‑quantitative scores ranging from 0 to 3 (see supplementary Table S1) (Agarwal et al., 2012; Carlisle et al., 2014).
Renal function monitoring
Biochemical analyses of serum were performed to determine the concentrations of uric acid, cystatin C, and creatinine. Biochemical parameter levels were measured by the RT-9200 automatic biochemical analyzer (Rayto Life and Analytical Sciences Co., Ltd. ShenZhen, P.R.
China).
Renal ion concentrations assay
The concentrations of Na+, K+, Cl−, Ca2+, and Mg2+ were measured in renal tissue. All kits were manufactured by Nanjing Jiancheng Bioengineering Institute (China), and the corresponding item numbers are C002-1-1, C001-2-1, C003-2-1, C004-2-1 and C005-1-1 in sequence.
Renal ATPase activity assay
Renal tissue homogenates were assayed for Na+-K+-ATPase, Ca2+-Mg2+-ATPase, Ca2+-ATPase and Mg2+-ATPase activity with the ATPase assay kit (A016-2-2, Nanjing Jiancheng Biotechnology Research Institute, China).
Quantitative real-time PCR (qRT–PCR) analysis
Kidney tissues were processed for total RNA extraction using a standard RNA isolation reagent (G3013-100 mL, Servicebio, Wuhan, China) and reverse-transcribed into cDNA with HiScript® II. qPCR was performed using the QRT SuperMix (+gDNA wipe) kit (Vazyme, Nanjing, China). The expression levels of target genes were measured by quantitative real-time PCR. ChamQ Universal SYBR qPCR Master Mix, a product of Vazyme Biotech (Nanjing, China), was used as the reaction reagent, and all amplification procedures were performed on a qTOWER® system supplied by Analytik Jena (Germany).
For the normalization of gene expression data, β-actin was set as the internal reference gene. All data calculation and analysis were carried out following the 2-ΔΔCt method (see supplementary Table S2).
Western blotting analysis
Total protein samples were isolated from renal tissues using RIPA lysis buffer and quantified with a BCA kit. Western blot testing was conducted based on the methods described in our prior research (Wang et al., 2024; Zhang et al., 2024). Protein bands were manufactured by means of the Amersham Imager 600 instrument made by GE (Switzerland), and the quantification was finished with ImageJ analysis software.
Primary antibodies against AQP1, AQP3, AQP4 and AQP5 were purchased from Wanleibio Company (Shenyang, China). The anti-β-actin antibody was obtained from Servicebio in Wuhan, China. The secondary antibodies included HRP-labeled Goat Anti-Rabbit IgG (H+L) and HRP-labeled Goat Anti-Mouse IgG (H+L), both of which were supplied by Servicebio (Wuhan, China).
Statistical analysis
Statistical analysis was performed using one-way ANOVA with GraphPad Prism software. Data are presented as mean ± SEM. Asterisks (*P < 0.05, **P < 0.01) represent remarkable variations between Con group and ATR treatment group.
Pound signs (#P < 0.05, ##P < 0.01) reflect notable statistical differences between ATR group and ATR + Se group.
Results
Effects of Yeast-Se on ATR-induced renal histopathological and functional impairment
To explore the protective impacts of Yeast-Se against ATR-triggered kidney injury, the histopathological morphology of the kidney was analyzed. Histopathological analysis of quail kidney tissues stained with H&E was performed. The kidney of Con group showed no significant pathological abnormalities, with clear and intact renal tubular structures, well-defined glomeruli, and regular morphology.
While, the kidney of Con ATR group exhibited pronounced pathological damage, including glomerular swelling and congestion, mesangial proliferation, vacuolar lesions. The ATR group showed edema of renal tubular epithelial cells with blurred cells with blurred cell boundaries. ATR + Se group showed reduced glomerular volume, fewer vacuoles, more distinct nuclei in renal tubular epithelial cells, and clearer cellular boundaries compared with ATR group (Fig.
1A and B). Compared with ATR treatment group, quails in ATR and Yeast-Se co-treatment group exhibited an obvious alleviation of kidney injury and a decreased histopathological score (P < 0.05 or P < 0.01) (Fig. 1C and D).
Biochemical analysis of serum demonstrated that the levels of uric acid, cystatin C, and creatinine were markedly increased in ATR group compared with Con group (P < 0.01). However, supplementation with Yeast-Se significantly reduced the levels of uric acid, cystatin C, and creatinine compared with ATR group (P < 0.01) (Fig. 1E-G).
The above findings demonstrated that ATR triggered structural and functional renal damage, and this impairment could be effectively alleviated by Yeast-Se supplementation.Fig. 1Effects of Yeast-Se on ATR-induced renal histopathological and functional impairment. (A) Macroscopic appearance of kidneys.
(B) Kidney H&E staining (400 ×). (C) Tubular damage score. (D) Glomerular damage score.
(E) Serum cystatin C levels. (F) Serum uric acid levels. (G) Serum creatinine levels.
Data are expressed as mean ± SEM (n = 3). Comparisons versus the Con group are indicated by asterisks: *P < 0.05, **P < 0.01. Comparisons between the ATR and ATR + Se groups are indicated by hash signs: # P < 0.05, ## P < 0.01.
Effects of Yeast-Se on ATR-induced disturbance of Na⁺, K⁺, and Cl⁻ homeostasis
ATR exposure induced a notable increase in the renal contents of Na⁺ and Cl⁻ compared with Con group (P < 0.01). However, the levels of Na⁺ and Cl⁻ in the ATR + Yeast-Se co treatment group were significantly lower than those in the ATR exposure group (P < 0.01) (Fig. 2A and B).
The renal K⁺ level was markedly decreased in ATR group compared with Con group (P < 0.01), while it was notably increased in ATR + Se group compared with ATR group (P < 0.05) (Fig. 2C).Fig. 2Effects of Yeast-Se on ATR-induced disturbance of Na⁺, K⁺, and Cl⁻ homeostasis.
(A) The content of Na+ in renal tissue. (B) The content of t of Cl- in renal tissue. (C) The content of K+- in renal tissue.
(D) The activity of Na⁺-K⁺-ATPase in the kidney. (E) Na+-K+-ATPase associated subunits relative mRNA expression. Data are expressed as mean ± SEM (n = 3).
Comparisons versus the Con group are indicated by asterisks: *P < 0.05, **P < 0.01. Comparisons between the ATR and ATR + Se groups are indicated by hash signs: # P < 0.05, ## P < 0.01.
Renal Na⁺-K⁺-ATPase activity was markedly decreased in ATR group in comparison with Con group (P < 0.01). However, ATR + Se group exhibited a notable upregulation in Na⁺-K⁺-ATPase activity compared with ATR treatment group (P < 0.01) (Fig. 2D).
Furthermore, in comparison with Con group, exposure to ATR significantly decreased the mRNA expression levels of Na⁺-K⁺-ATPase subunits (ATP1A1, ATP1A2, ATP1A3, ATP1B3, and ATP1B4) (P < 0.01). However, Yeast-Se supplementation notably elevated the transcriptional levels of these subunits compared with ATR group (P < 0.01) (Fig. 2E).
Effects of Yeast-Se on ATR-induced disturbance of Ca²⁺ and Mg²⁺ homeostasis
As shown in Fig. 3A, ATR exposure significantly decreased renal Ca²⁺ concentration compared with Con group, while Yeast-Se supplementation significantly restored Ca²⁺ levels suppressed by ATR exposure (P < 0.01). Exposure to ATR boosted the renal Mg²⁺ concentration compared with Con group (P < 0.01), while Yeast-Se supplementation significantly decreased Mg²⁺ concentration in comparison to ATR group (P < 0.01) (Fig.
3B).Fig. 3Effects of Yeast-Se on ATR-induced disturbance of Ca²⁺ and Mg²⁺ homeostasis. (A) The content of Ca2+ in renal tissue.
(B) The content of Mg2+ in renal tissue. (C) The activity of Ca²⁺-ATPase in the kidney. (D) The activity of Mg²⁺-ATPase in the kidney.
(E) The activity of Ca²⁺-Mg²⁺-ATPase in the kidney. (F) Ca²⁺ transfer channel associated subunits relative mRNA expression. Data are expressed as mean ± SEM (n = 3).
Comparisons versus the Con group are indicated by asterisks: *P < 0.05, **P < 0.01. Comparisons between the ATR and ATR + Se groups are indicated by hash signs: # P < 0.05, ## P < 0.01.
The activities of renal Ca²⁺-ATPase, Mg²⁺-ATPase and Ca²⁺-Mg²⁺-ATPase were significantly higher in ATR group than those in Con group (P < 0.01). Compared to ATR group, supplementing with Yeast-Se significantly decreased these activities (P < 0.05 or P < 0.01) (Fig. 3C–E).
The mRNA expression levels of Ca²⁺-ATPase subunits (ATP2A1, ATP2A2, ATP2A3), Na⁺/Ca²⁺ exchanger subunits (SLC8A1, SLC8A2, SLC8A3, SLC8B1), and Na⁺/Ca²⁺-K⁺ exchanger subunits (SLC24A1, SLC24A2) exhibited a substantial decline in ATR group compared with Con group (P < 0.01). Yeast-Se administration significantly markedly alleviated the alterations of Ca²⁺-ATPase subunits (ATP2A1, ATP2A2, ATP2A3), Na⁺/Ca²⁺ exchanger subunits (SLC8A1, SLC8A2), and the Na⁺/Ca²⁺-K⁺ exchanger subunit (SLC24A1) induced by ATR (P < 0.01) (Fig. 3F).
Effects of Yeast-Se on ATR-induced disturbance of AQPs expression
In comparison with Con group, ATR exposure markedly decreased the protein expression levels of AQP3 and AQP4, while remarkably elevated the protein levels of AQP1 and AQP5 (P < 0.05 or P < 0.01). While, Yeast-Se supplementation notably upregulated the levels of AQP3 and AQP4, while downregulating AQP1 and AQP5 in comparison with ATR group (P < 0.05 or P < 0.01) (Fig. 4A–D).
The mRNA expression levels of AQP1 and AQP11 were significantly elevated, while AQP2, AQP7, and AQP12 were markedly decreased in ATR group compared with Con group (P < 0.05 or P < 0.01). However, ATR + Se group significantly alleviated the alteration AQP1, AQP2, AQP5, AQP7, and AQP11. AQP12 gene expression level caused by ATR (P < 0.05 or P < 0.01) (Fig.
4E). These collective findings demonstrated that effectively reversed ATR-induced disturbances in renal AQPs expression.Fig. 4Effects of Yeast-Se on ATR-induced disturbance of AQPs expression.
(A) The protein expression of AQP1. (B) The protein expression of AQP3. (C) The protein expression of AQP4.
(D) The protein expression of AQP5. (E) The mRNA expression of AQPs in the kidney. Data are expressed as mean ± SEM (n = 3).
Comparisons versus the Con group are indicated by asterisks: *P < 0.05, **P < 0.01. Comparisons between the ATR and ATR + Se groups are indicated by hash signs: # P < 0.05, ## P < 0.01.
Discussion
ATR is recognized as one of the most widely applied herbicides in agricultural production worldwide. ATR persists in soil and water systems, leading to bioaccumulation in the food chain and posing significant health threats with non-target organisms (Dehghani et al., 2022; Gagneten et al., 2023). Kidney serves as a key organ for regulating water-salt balance and ionic homeostasis, and is also a potential target organ of ATR, and disruption of ion homeostasis is a primary toxic effect of ATR-induced nephrotoxicity.
In this study, ATR disrupted renal structure and function, induced ion imbalance, suppressed ATPase activities, and dysregulated AQPs expression. However, Yeast-Se supplementation reversed ATR‑induced renal damage and ion disturbances by modulating ATPase activities and AQPs expression, thereby protecting against ATR-induced nephrotoxicity.
Ionic homeostasis is critical for maintaining cellular integrity and physiological function (Zhai et al., 2020). Kidney serves a critical function in maintaining systemic ionic balance, and disorder of ionic homeostasis serves as a key mechanism underlying nephrotoxicity (Caiati et al., 2025). Disruption of renal ion homeostasis, particularly of Na⁺, K⁺, Ca²⁺, and Mg²⁺, can lead to cellular dysfunction, apoptosis, and organ failure (Katopodis et al., 2022).
It has been documented that exposure to ATR can cause ionic disorder in the liver and heart on quails (Lin et al., 2016a, 2016b). In this study, ATR exposure caused pronounced disturbances in renal ion homeostasis in quails. Specifically, ATR significantly elevated renal Na⁺, Cl⁻ and Mg²⁺ content, and reduced K⁺ and Ca²⁺ levels, which are similar to previous research findings (Guo et al., 2021).
Therefore, ion homeostasis disorder is a main toxic effect of ATR exposure among multiple organs. ATPases are essential for active ion transport across cell membraned, and inhibition of ATPases can disrupt electrochemical gradients and cellular signaling (Fedosova et al., 2021). Earlier investigations have confirmed that ion channels dysfunction is intrinsically linked to renal injury (Li et al., 2025).
ATR exposure can cause hepatic ion homeostasis disorder via suppressing ATPases activity (Wang et al., 2024). In the current study, ATR suppressed Na⁺-K⁺-ATPase activity and downregulated the mRNA expression levels of Na⁺-K⁺-ATPase subunits (ATP1A1, ATP1A2, ATP1A3, ATP1B3, and ATP1B4), which fully explains why ATR exposure increased renal Na⁺ and Cl⁻ and decreased K⁺ levels in the kidney. At the same time, ATR increased the activities of Ca²⁺-ATPase, Mg²⁺-ATPase, and Ca²⁺-Mg²⁺-ATPase, while reducing the mRNA expression of Ca²⁺-ATPase subunits (ATP2A1, ATP2A2, and ATP2A3), Na⁺/Ca²⁺ exchangers (SLC8A1, SLC8A2, SLC8A3, and SLC8B1), and Na⁺/Ca²⁺-K⁺ exchangers (SLC24A1 and SLC24A2), which accounted for the alterations of renal Ca²⁺ and Mg²⁺ levels induced by ATR.
Hence, it can be speculated that ATR-induced ionic disturbance is confirmed to be associated with changes in ATPases activity. However, dietary supplementation with Yeast-Se effectively reversed ATR-induced renal ion imbalances by regulating ATPase activities.
Traditionally, AQPs are considered to be transmembrane water channels that mediate water transport along osmotic gradients (Hachez and Chaumont, 2010). However, accumulating evidence indicates that AQPs also participate in regulating ion homeostasis and signaling pathways (Login and Nejsum, 2023). According to a recent comprehensive review, AQPs play essential roles in maintaining tubular epithelial integrity by mediating cell proliferation, migration, and apoptosis (Venkatesan et al., 2026).
AQP3 and AQP4 are known to be involved in water and glycerol transport, which have been linked to renal dysfunction in response to environmental toxicants (He and Yang, 2019). In the context of nephrotoxicity, renal AQPs dysregulation contributed to disturbances in water and ionic balance (Nielsen et al., 2008). In the current investigation, ATR significantly downregulated AQP3 and AQP4 at the protein level, and upregulated at both mRNA and protein levels of AQP1 and AQP5.
In addition, ATR also remarkably disrupt the mRNA expression levels of AQP2, AQP7, AQP11, and AQP12. It has been widely documented in earlier investigations that that ATR induces necroptosis of tubular epithelial cells in kidney (Wang et al., 2026) and disrupts AQPs expression (Jiang et al., 2021), which is in agreement with recent study that AQP2 plays a critical role in regulating renal Na+ and Ca2+ handling in mice exposed to ATR (Guo et al., 2021). Thus, the dysregulation of AQPs expression is proposed as a critical factor in renal ion disorder induced by ATR.
Yeast-Se was shown to alleviate ATR-induced ionic disturbance by modulating AQPs expression in liver of quails (Wang et al., 2024). Yeast-Se exerted a regulatory effect on renal AQP expression, with upregulation of AQP3 and AQP4 and downregulation of AQP1 and AQP5. Yeast-Se exhibits potent anti-inflammatory and antioxidant properties (Yang and Yang, 2023), and whether it alleviates ATR-induced AQPs dysregulation through these effects remains to be explored in subsequent research.
In the present study, ATR exposure disrupted ATPase activities and subunit transcription, causing significant renal ion disorder and subsequent renal injury. Additionally, AQPs are functionally coupled with ATPases, which expression were also dysregulated induced by ATR. Hence, it can be concluded that ATR-induced ionic disturbance is attributed to alterations in ATPases activity and AQPs expression in kidney.
In this study, Yeast-Se mitigates ATR-induced nephrotoxicity by regulating ATPases activity and AQPs expression. This study extends these findings to kidney and provides new evidence for the renal protective role of Yeast-Se. In spite of these insights, a number of inherent limitations need to be recognized.
First, the dose of ATR (500 mg/kg B.W.) used in this study was higher than realistic environmental exposure levels and was selected based on previous toxicological studies to reproduce consistent renal injury and ionic disturbance in quails. Future studies using environmentally relevant low-dose exposures are warranted to validate the translational relevance of our findings. Second, the in vivo nature of this study precludes direct causal inference between AQPs expression and ion transport.
Future studies using AQP-knockdown or overexpression models in renal tubular cells would help clarify their functional roles. Third, although multiple AQPs isoforms were measured, their subcellular localization and post-translational modifications in response to ATR and Yeast-Se remain to be investigated. In spite of these insights, a number of inherent limitations need to be recognized.
First, the in vivo nature of this study precludes direct causal inference between AQPs expression and ion transport. Future studies using AQP-knockdown or overexpression models in renal tubular cells would help clarify their functional roles. Second, although multiple AQPs isoforms were measured, their subcellular localization and post-translational modifications in response to ATR and Yeast-Se remain to be investigated.
Conclusion
Yeast-Se markedly mitigated ATR-induced renal damage by restoring both ATPases function and the transcription of their subunits, and normalizing AQPs expression, to corrected ionic imbalance. Collectively, the above findings enhance our comprehension of nephrotoxicity induced by ATR, and meanwhile support the promising application of Yeast-Se as a dietary intervention against herbicide-induced renal damage in poultry.
Author Contributions
Yanan Li: Substantial contributions to study design, experiments, data collection and analysis; drafted the manuscript; approved the final version; accountable for the work. Peilin Wang: Contributed substantially to experiments, data collection and analysis; critically revised the manuscript; approved the final version; and takes accountability for the integrity of this work. Yiming Wang: Contributed substantially to data analysis and interpretation; critically revised the manuscript; approved the final version; and takes accountability for the integrity of this work.
Xuebing Wang: Contributed substantially to data collection and analysis and provided resources; critically revised the manuscript; approved the final version; and takes accountability for the integrity of this work. Xu Yang: Contributed substantially to study conception and design, funding acquisition and project supervision; critically revised the manuscript; approved the final version; and takes accountability for the integrity of this work. Cong Zhang: Contributed substantially to study conception and design, funding acquisition, resource provision and supervision; critically revised the manuscript; approved the final version; and takes accountability for the integrity of this work.
Funding
This work was supported by 10.13039/100007471Henan Provincial Science and Technology Research and Development Plan Joint Fund (252301420012) and 10.13039/501100012245Henan Province Science and Technology Key Project (262102311264).
Disclosures
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.