高脂饮食与低剂量TCDD联合作用对雌性肥胖易感大鼠肝脏脂质代谢的影响

doi:10.3969/j.issn.1006-7795.2023.06.019

摘要/Abstract

摘要： 目的探讨高脂饮食（high fat diet，HFD)引起的肥胖与低剂量持久性有机污染物——2,3,7,8-四氯二苯并二噁英（2,3,7,8-tetrachlorodibenzo-p-dioxin, TCDD)联合作用对肝脏脂质代谢的影响。方法采用2×2析因设计，研究HFD与低剂量TCDD（10 ng·kg^－1·d^－1)联合作用对10周龄肥胖易感（obesity-prone，OP)雌性大鼠肝脏脂质代谢的影响。4组实验动物分别为：普通饮食无TCDD处理组（Cont+TCDD 0)、普通饮食低剂量TCDD处理组（Cont+TCDD 10)、高脂饮食无TCDD处理组（HFD+TCDD 0)及高脂饮食低剂量TCDD处理组（HFD+TCDD 10)。油红O染色观察肝脏脂肪沉积，酶法测定肝脏超氧化物歧化酶（superoxide dismutase, SOD)和谷胱甘肽过氧化物酶（glutathione peroxidase, GSH-PX)的活性，实时荧光定量聚合酶链式反应（real-time fluorescence quantitative polymerase chain reaction, RT- qPCR)法检测肝脏脂质代谢相关基因mRNA表达。结果 Cont+TCDD 0组、Cont+TCDD 10组、HFD+TCDD 0组及HFD+TCDD 10组大鼠体质量分别为（301.29±18.47)g、（312.16±20.45)g、（358.20±27.11)g及（373.83±26.50)g （F=7.43，P<0.01)；肝脏油红O阳性区域比值分别为（1.26±0.16)%、（1.66±0.50)%，（9.06±1.54)%、（32.56±2.66)% （F=11.32，P<0.01)，肝脏SOD酶活性分别为：（1.54±0.06)U/mg、（1.41±0.15)U/mg、（1.25±0.08)U/mg及（0.76±0.11) U/mg（F=4.31，P<0.05)；肝脏GSH-PX酶活性分别为：（1 511.00±69.30)U/mg、（1 409.00±60.81)U/mg、（1 232.00±65.05)U/mg及（965.00±83.44)U/mg（F=3.91，P<0.05)。对上述指标进行交互作用分析显示，高脂饮食与低剂量TCDD联合作用使肥胖易感的雌性SD大鼠体质量（F=6.33，P<0.05)及肝脏脂肪积聚（F=12.51，P<0.01)出现协同增加，肝脏抗氧化酶类SOD酶活性（F=4.15，P<0.05)和GPX酶活性（F=3.97，P<0.05)协同降低。对12种肝脏脂质代谢相关基因mRNA表达水平检测发现，高脂饮食与低剂量TCDD联合作用增加三酰甘油脂肪酶的限速酶（adipose triglyceride lipase，ATGL)（F=3.69，P<0.05)及脂肪酸转位酶（cluster of differentiation 36，CD36)的mRNA表达（F=5.58，P<0.01)，抑制激素敏感脂肪酶（hormone-sensitive lipase，HSL)的mRNA表达（F=4.01，P<0.05)，其余9种基因表达差异无统计学意义（P＞0.05)。结论高脂饮食与低剂量TCDD联合作用可能协同增加肥胖诱导雌性大鼠肝脏脂质代谢异常。

关键词: 肥胖症, 2,3,7,8-四氯二苯并二噁英, 高脂膳食, 氧化应激

Abstract: Objective To study the effect of combination of high fat diet (HFD) induced obesity and low dose persistent organic pollutants 2,3,7,8-tetrachlorodibenzo-p-dioxin （TCDD) on liver lipid metabolism. Methods By 2×2 factorial design, the effects of combination of HFD and low dose TCDD (10 ng·kg^－1·d^－1) on liver lipid metabolism in obesity-prone (OP) 10-week-old female rats were studied. The four groups of experimental animals were: the group treated with no TCDD in chow diet (Cont+TCDD 0), the group treated with low dose TCDD in chow diet (Cont+TCDD 10), and the group high fat diet without TCDD (HFD+TCDD 0) and high fat diet group treated with low dose TCDD (HFD+TCDD 10). Lipid deposition in liver was observed by oil red O staining. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) in liver were determined by enzyme method. mRNA expression of genes related to lipid metabolism in liver was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Results The body weight of rats in Cont+TCDD 0 group, Cont+TCDD 10 group, HFD+TCDD 0 group and HFD+TCDD 10 group were (301.29±18.47) g, (312.16±20.45) g, (358.20±27.11) g and (373.83±26.50) g, respectively (F=7.43, P<0.01), liver oil red O positive area ratios were （1.26±0.16)%,（1.66±0.50)%，（9.06±1.54)%,（32.56±2.66)% (F=11.32, P<0.01), liver SOD activity were (1.54±0.06) U/mg, (1.41±0.15) U/mg, (1.25±0.08) U/mg and (0.76±0.11) U/mg (F=4.31, P<0.05), respectively. Liver GSH-PX activity were Liver GSH-PX activity were (1 511.00±69.30) U/mg, (1 409.00±60.81) U/mg, (1 232.00±65.05) U/mg and (965.00±83.44) U/mg (F=3.91, P<0.05). The interaction analysis of the above indexes suggested that the combination of high fat diet and low dose TCDD resulted in synergistic increases in the body weight (F=6.33, P<0.05) and liver fat accumulation (F=12.51, P<0.01) of obese-prone female SD rats, and the activities of antioxidant enzymes such as SOD in liver (F=4.15, P<0.05) and GSH-PX enzyme activity (F=3.97, P<0.05) were decreased synergistically. Analysis on mRNA expression levels of 12 genes related to lipid metabolism in liver showed that high fat diet combined with low dose TCDD increased adipose lipase (Atgl) (F=3.69, P<0.05), mRNA expression of cluster of differentiation 36 (CD36) (F=5.58, P<0.01), inhibited the mRNA expression of hormone sensitive lipase (Hsl) (F=4.01, P<0.05), and there was no significant difference in the expression of other 9 genes (P>0.05). Conclusion The combination of high fat diet and low dose TCDD can synergistically increase the abnormal liver lipid metabolism in female obesity-prone female SD rats.

Key words: obesity, 2,3,7,8-tetrachlorodibenzo-p-dioxin, diet high fat, oxidative stress

中图分类号:

刘月, 朱丹, 洪煜婧, 孙文星, 徐广飞. 高脂饮食与低剂量TCDD联合作用对雌性肥胖易感大鼠肝脏脂质代谢的影响[J]. 首都医科大学学报, 2023, 44(6): 1029-1035.

Liu Yue, Zhu Dan, Hong Yujing, Sun Wenxing, Xu Guangfei. Effects of high fat diet with low dose TCDD on hepatic lipid metabolism in female obesity-prone SD rats[J]. Journal of Capital Medical University, 2023, 44(6): 1029-1035.

参考文献

［1］Levin B E, Triscari J, Sullivan A C. Relationship between sympathetic activity and diet-induced obesity in two rat strains［J］. Am J Physiol, 1983, 245(3): R364-R371.
［2］Sinha-Hikim A P, Sinha-Hikim I, Friedman T C. Connection of nicotine to diet-induced obesity and non-alcoholic fatty liver disease: cellular and mechanistic insights［J］. Front Endocrinol (Lausanne), 2017, 8: 23.
［3］Jurgelewicz A, Nault R, Harkema J, et al. Characterizing the impact of simvastatin co-treatment of cell specific TCDD-induced gene expression and systemic toxicity［J］. Sci Rep, 2023, 13(1): 16598.
［4］Li C Y, Liu Y Y, Dong Z, et al. TCDD promotes liver fibrosis through disordering systemic and hepatic iron homeostasis［J］. J Hazard Mater, 2020, 395: 122588.
［5］Angrish M M, Mets B D, Jones A D, et al. Dietary fat is a lipid source in 2,3,7,8-tetrachlorodibenzo-ρ-dioxin (TCDD)-elicited hepatic steatosis in C57BL/6 mice［J］. Toxicol Sci, 2012, 128(2): 377-386.
［6］Deierlein A L, Rock S, Park S. Persistent endocrine-disrupting chemicals and fatty liver disease［J］. Curr Environ Health Rep, 2017, 4(4): 439-449.
［7］Hakkarainen K, Rantakokko P, Koponen J, et al. Persistent organic pollutants associate with liver disease in a Finnish general population sample［J］. Liver Int, 2023, 43(10): 2177-2185.
［8］Eslam M, Newsome P N, Sarin S K, et al. A new definition for metabolic dysfunction-associated fatty liver disease: an international expert consensus statement［J］. J Hepatol, 2020, 73(1): 202-209.
［9］Türkez H, Geyikog∨lu F, Yousef M I, et al. Propolis alleviates 2,3,7,8-Tetrachlorodibenzo-p-dioxin-induced histological changes, oxidative stress and DNA damage in rat liver［J］. Toxicol Ind Health, 2013, 29(8): 677-685.
［10］Gao L, Mutlu E, Collins LB, et al.?? DNA product formation in female Sprague-Dawley rats following polyhalogenated aromatic hydrocarbon (PHAH) exposure［J］. Chem Res Toxicol,2017, 30(3):794-803.
［11］Hassoun E A, Li F, Abushaban A, et al. The relative abilities of TCDD and its congeners to induce oxidative stress in the hepatic and brain tissues of rats after subchronic exposure［J］. Toxicology, 2000, 145(2/3): 103-113.
［12］刘小方, 刘潇, 曹文成, 等. 湖北省某地区居民血清中多氯联苯的负荷水平调查［J］. 中华预防医学杂志, 2021, 55(6): 792-796.
［13］Lee J H, Wada T, Febbraio M, et al. A novel role for the dioxin receptor in fatty acid metabolism and hepatic steatosis［J］. Gastroenterology, 2010, 139(2): 653-663.
［14］Lee J, Song J H, Park J H, et al. Dnmt1/Tet2-mediated changes in Cmip methylation regulate the development of nonalcoholic fatty liver disease by controlling the Gbp2-Pparγ-CD36 axis［J］. Exp Mol Med, 2023, 55(1): 143-157.
［15］Habets D D J, Coumans W A, Voshol P J, et al. AMPK-mediated increase in myocardial long-chain fatty acid uptake critically depends on sarcolemmal CD36［J］. Biochem Biophys Res Commun, 2007, 355(1): 204-210.

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