Eti Nazmin Akter, Flor Susanne, Iqbal Khursheed, Scott Regan L, Klenov Violet E, Gibson-Corley Katherine N, Soares Michael J, Ludewig Gabriele, Robertson Larry W
Interdisciplinary Graduate Program in Human Toxicology, University of Iowa, Iowa City, IA, United States.
Department of Radiation Oncology, University of Iowa, Iowa City, IA, United States.
Toxicology. 2022 Jan 30;466:153054. doi: 10.1016/j.tox.2021.153054. Epub 2021 Nov 27.
The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor involved in the regulation of biological responses to more planar aromatic hydrocarbons, like TCDD. We previously described the sequence of events following exposure of male rats to a dioxin-like polychlorinated biphenyl (PCB) congener, 3,3',4,4',5-pentachlorobiphenyl (PCB126), that binds avidly to the AhR and causes various types of toxicity including metabolic syndrome, fatty liver, and disruption of energy homeostasis. The purpose of this study was, to investigate the role of AhR to mediate those toxic manifestations following sub-acute exposure to PCB126 and to examine possible sex differences in effects. For this goal, we created an AhR knockout (AhR-KO) model using CRISPR/Cas9. Comparison was made to the wild type (WT) male and female Holtzman Sprague Dawley rats. Rats were injected with a single IP dose of corn oil vehicle or 5 μmol/kg PCB126 in corn oil and necropsied after 28 days. PCB126 caused significant weight loss, reduced relative thymus weights, and increased relative liver weights in WT male and female rats, but not in AhR-KO rats. Similarly, significant pathologic changes were visible which included necrosis and regeneration in female rats, micro- and macro-vesicular hepatocellular vacuolation in males, and a paucity of glycogen in livers of both sexes in WT rats only. Hypoglycemia and lower IGF1, and reduced serum non-esterified fatty acids (NEFAs) were found in serum of both sexes of WT rats, low serum cholesterol levels only in the females, and no changes in AhR-KO rats. The expression of genes encoding enzymes related to xenobiotic metabolism (e.g. CYP1A1), gluconeogenesis, glycogenolysis, and fatty acid oxidation were unaffected in the AhR-KO rats following PCB126 exposure as opposed to WT rats where expression was significantly upregulated (PPARα, females only) or downregulated suggesting a disrupted energy homeostasis. Interestingly, Acox2, Hmgcs, G6Pase and Pc were affected in both sexes, the gluconeogenesis and glucose transporter genes Pck1, Glut2, Sds, and Crem only in male WT-PCB rats. These results show the essential role of the AhR in glycogenolysis, gluconeogenesis, and fatty acid oxidation, i.e. in the regulation of energy production and homeostasis, but also demonstrate a significant difference in the effects of PCB126 in males verses females, suggesting higher vulnerability of glucose homeostasis in males and more changes in fatty acid/lipid homeostasis in females. These differences in effects, which may apply to more/all AhR agonists, should be further analyzed to identify health risks to specific groups of highly exposed human populations.
芳基烃受体(AhR)是一种配体激活的转录因子,参与对更多平面芳香烃(如TCDD)的生物反应调节。我们之前描述了雄性大鼠暴露于一种二噁英样多氯联苯(PCB)同系物3,3',4,4',5-五氯联苯(PCB126)后的一系列事件,该同系物与AhR紧密结合并导致包括代谢综合征、脂肪肝和能量稳态破坏在内的各种毒性。本研究的目的是调查AhR在亚急性暴露于PCB126后介导这些毒性表现中的作用,并检查可能存在的性别差异。为实现这一目标,我们使用CRISPR/Cas9创建了一个AhR基因敲除(AhR-KO)模型。将其与野生型(WT)雄性和雌性霍尔兹曼·斯普拉格·道利大鼠进行比较。大鼠单次腹腔注射玉米油载体或玉米油中5 μmol/kg的PCB126,28天后进行尸检。PCB126导致WT雄性和雌性大鼠体重显著减轻、相对胸腺重量降低以及相对肝脏重量增加,但在AhR-KO大鼠中未出现这种情况。同样,在WT大鼠中可见明显的病理变化,包括雌性大鼠的坏死和再生、雄性大鼠的微泡和大泡性肝细胞空泡化,以及仅在WT大鼠两性肝脏中糖原缺乏。WT大鼠两性血清中均发现低血糖、较低的IGF1以及血清非酯化脂肪酸(NEFA)降低,仅雌性大鼠血清胆固醇水平降低,而AhR-KO大鼠无变化。与WT大鼠中表达显著上调(仅雌性中的PPARα)或下调表明能量稳态破坏相反,AhR-KO大鼠在暴露于PCB126后,编码与异源物代谢相关酶(如CYP1A1)、糖异生、糖原分解和脂肪酸氧化的基因表达未受影响。有趣的是,Acox2、Hmgcs、G6Pase和Pc在两性中均受影响,糖异生和葡萄糖转运蛋白基因Pck1、Glut2、Sds和Crem仅在雄性WT-PCB大鼠中受影响。这些结果表明AhR在糖原分解、糖异生和脂肪酸氧化中起着关键作用,即在能量产生和稳态调节中起关键作用,但也表明PCB126对雄性和雌性的影响存在显著差异,表明雄性对葡萄糖稳态的易感性更高,而雌性在脂肪酸/脂质稳态方面变化更多。这些影响差异可能适用于更多/所有AhR激动剂,应进一步分析以确定对高暴露特定人群组的健康风险。