基于Nrf2-HO-1通路探讨叶黄素对锰暴露大鼠颅脑神经损伤的保护作用

职业与健康 ›› 2025, Vol. 41 ›› Issue (9): 1180-1186.

基于Nrf2-HO-1通路探讨叶黄素对锰暴露大鼠颅脑神经损伤的保护作用

丁查文^a, 乔红^b

重钢总医院a神经内科,b眼科,重庆 400000

收稿日期:2024-08-02 修回日期:2024-08-12 出版日期:2025-05-01 发布日期:2025-12-12
通信作者: 乔红,主治医师,E-mail：qiaohong8544535@126.com
作者简介:丁查文,男,主治医师,主要从事脑血管神经疾病的研究工作。
基金资助:
重庆市医学科研项目（2019063）

Exploring the protective effect of lutein on cranial nerve damage in rats exposed to manganese based on the Nrf2-HO-1 pathway

DING Chawen^a, QIAO Hong^b

a Department of Neurology,b Department of Ophthalmology,Chonggang General Hospital,Chongqing 400000,China

Received:2024-08-02 Revised:2024-08-12 Online:2025-05-01 Published:2025-12-12
Contact: QIAO Hong,Attending physician,E-mail：qiaohong8544535@126.com

摘要/Abstract

摘要： 目的基于NF-E2相关因子2（nf-e2-related factor 2,Nrf2/血红素氧酶-1（heme oxygenase-1,HO-1）通路探讨叶黄素对锰暴露大鼠颅脑神经损伤的保护作用。方法 50只SD大鼠,雌雄各半分成对照组、模型组、叶黄素低、中、高剂量组（40.0、80.0、160.0 mg/kg）;除对照组外,其余各组大鼠每日予以硫酸锰溶液（30 mg/kg）灌胃,持续 3个月,实验期间,叶黄素低、中、高剂量大鼠每日予以相应剂量的叶黄素灌胃,对照组、模型组给予等体积的生理盐水灌胃,试验结束后测定大鼠神经功能指标、神经细胞凋亡水平、海马组织超氧化物歧化酶（superoxide dismutase,SOD）、谷胱甘肽（glutathione,r-glutamyl cysteingl+glycine,GSH）、乳酸脱氢酶（lactate dehydrogenase,LDH）、活性氧（reactive oxygen species,ROS）、丙二醛（malondialdehyde, MDA）;实时荧光逆转录（real-time fluorescence reverse transcription,RT-PCR）法及蛋白印记法测定海马组织Nrf2、醌氧化还原酶-1（quinone oxidoreductase-1,NQO-1）、HO-1水平。结果锰暴露染毒组大鼠路径长度、斜面测试评分、海马组织SOD、GSH水平、海马组织Nrf2、NQO-1、HO-1 mRNA蛋白水平低于对照组（4.28±0.41 vs 0.86±0.07、4.30±0.38 vs 0.53±0.06、4.03±0.41 vs 0.53±0.05,1.08±0.10 vs 0.28±0.03、1.18±0.12 vs 0.19±0.03、0.19±0.03 vs 0.21±0.02）,Longa神经系统评分、改良神经系统严重程度评分（modified neurological deficit score,mNSS）、海马TdT介导的dUTP缺口末端标记（TdT-mediated dUTP nick end labeling,TUNEL）阳性细胞数、海马组织LDH、ROS、MDA水平高于对照组,差异均有统计学意义（均P<0.05）;低中高剂量的叶黄素干预治疗后,各叶黄素剂量组大鼠路径长度、斜面测试评分、海马组织SOD、GSH水平、海马组织Nrf2、NQO-1、HO-1 mRNA蛋白水平高于锰暴露染毒组（0.86±0.07 vs 1.26±0.10、1.56±0.17、2.84±0.23,0.53±0.06 vs 1.22±0.11、1.64±0.20、 3.01±0.29,0.53±0.05 vs 1.18±0.14、1.82±0.19、3.14±0.30;0.28±0.03 vs 0.58±0.06、0.81±0.08、0.99±0.08,0.19±0.03 vs 0.31±0.04、0.63±0.07、1.03±0.09,0.21±0.02 vs 0.46±0.07、0.67±0.07、1.02±0.10,均P<0.05）,Longa神经系统评分、mNSS评分、海马TUNEL阳性细胞数、海马组织LDH、ROS、MDA水平低于锰暴露染毒组,差异均有统计学意义（均P<0.05）;且随着叶黄素剂量的逐渐增加,低中高剂量叶黄素组大鼠路径长度、斜面测试评分、海马组织SOD、GSH水平、海马组织Nrf2、NQO-1、HO-1 mRNA蛋白水平逐渐升高（1.26±0.10、1.56±0.17、2.84±0.23,1.22±0.111.64±0.20、3.01±0.29,1.18±0.14、1.82±0.19、3.14±0.30;0.58±0.06、0.81±0.08、0.99±0.08,0.31±0.04、0.63±0.07、1.03±0.09,0.46±0.07、0.67±0.07、1.02±0.10）,Longa神经系统评分、mNSS评分、海马TUNEL阳性细胞数、海马组织LDH、ROS、MDA水平逐渐降低,剂量-效应关系明显（P<0.05）。结论叶黄素对Mn暴露引起的颅脑神经损伤具有显著的治疗作用,其机制与叶黄素激活Mn暴露大鼠海马神经Nrf2-HO-1通路有关。

Abstract: Objective To explore the protective effect of lutein on cranial nerve injury in rats exposed to manganese based on the nf-e2-related factor 2（Nrf2）-heme oxygenase-1（HO-1） pathway. Methods Totally 50 SD rats were divided into half and half of both sexes：control group,model group,low,medium and high dose lutein groups（40.0,80.0,160.0 mg/kg）. Except for the control group, all other groups of rats were given manganese sulfate solution（30 mg/kg） by gavage every day for 3 months. During the experiment, rats with low,medium and high doses of lutein were given corresponding doses of lutein by gavage every day,the control group and model group were given equal volumes of normal saline by gavage. After the experiment,the rats' neurological function indicators,neuronal apoptosis levels,superoxide dismutase（SOD）,glutathione（GSH）,lactate dehydrogenase（LDH）,reactive oxygen species（ROS）,and malondialdehyde（MDA） in hippocampal tissue were measured. Real-time fluorescence reverse transcription（RT-PCR） and Western blot were used to measure the levels of Nrf2,quinone oxidoreductase-1（NQO-1）,and HO-1 in hippocampal tissue. Results The pathway length,slope test score,hippocampal tissue SOD and GSH levels,and hippocampal tissue Nrf2,NQO-1, and HO-1 mRNA protein levels of rats in the manganese exposure group were lower than those in the control group,Longa nervous system score,modified neurological deficit score（mNSS） score,TdT-mediated dUTP nick end labeling（TUNEL） positive cells in the hippocampus,hippocampal tissue LDH,ROS,and MDA levels were higher than those in the control group,and the differences were statistically significant（all P<0.05）. After intervention treatment with low,medium,and high doses of lutein,the pathway length,slope test scores,SOD and GSH levels in hippocampal tissue,and Nrf2,NQO-1,and HO-1 mRNA protein levels in hippocampal tissue in each lutein dose group were higher than those in the manganese exposure group（0.86±0.07 vs 1.26±0.10, 1.56±0.17,2.84±0.23;0.53±0.06 vs 1.22±0.11,1.64±0.20,3.01±0.29;0.53±0.05 vs 1.18±0.14,1.82±0.19,3.14±0.30;0.28±0.03 vs 0.58±0.06,0.81±0.08,0.99±0.08;0.19±0.03 vs 0.31±0.04,0.63±0.07,1.03±0.09;0.21±0.02 vs 0.46±0.07,0.67±0.07,1.02±0.10,all P<0.05）,Longa nervous system score,mNSS score,number of TUNEL-positive cells in hippocampus,and hippocampal tissue LDH,ROS,and MDA levels were lower than those in the manganese exposure group,and the differences were statistically significant（all P<0.05）,and with the gradual increase of lutein dosage,the pathway length,slope test scores,SOD and GSH levels in hippocampal tissue,Nrf2,NQO-1,and HO-1 mRNA protein levels in hippocampal tissue gradually increased（1.26±0.10,1.56±0.17,2.84±0.23;1.22±0.11,1.64±0.20,3.01±0.29;1.18±0.14,1.82±0.19,3.14±0.30;0.58±0.06,0.81±0.08,0.99±0.08;0.31±0.04, 0.63±0.07,1.03±0.09;0.46±0.07,0.67±0.07,1.02±0.10）,Longa nervous system score,mNSS score,number of TUNEL-positive cells in hippocampus,LDH,ROS,and MDA levels in hippocampal tissue gradually decreased,and the dose-effect relationship was significant（all P<0.05）. Conclusion Lutein has a significant therapeutic effect on cranial nerve damage caused by manganese exposure,and its mechanism is related to the activation of the Nrf2-HO-1 pathway in the hippocampus of manganese exposed rats by lutein.

Key words: Nrf2, HO-1, Lutein, Manganese, Cranial nerve injury

中图分类号:

R-332

丁查文, 乔红. 基于Nrf2-HO-1通路探讨叶黄素对锰暴露大鼠颅脑神经损伤的保护作用. [J]职业与健康, 2025, 41(9): 1180-1186.

DING Chawen, QIAO Hong. Exploring the protective effect of lutein on cranial nerve damage in rats exposed to manganese based on the Nrf2-HO-1 pathway. [J]OCCUPATION AND HEALTH, 2025, 41(9): 1180-1186.

参考文献

[1] VLASAK T,DUJLOVIC T,BARTH A.Manganese exposure and cognitive performance:A meta-analytical approach[J].Env Pollution,2023, 9(9):121884.
[2] CRISWELL S R,NIELSEN S S,FAUST I M,et al.Neuroinflammation and white matter alterations in occupational manganese exposure assessed by diffusion basis spectrum imaging[J].Neuro Toxicol,2023,97(14):25-33.
[3] KARAKURT Y,SÜLEYMAN H,KESKIN CIMEN F,et al.The effects of lutein on optic nerve injury induced by ethambutol and isoniazid:An experimental study[J].Cutan Ocul Toxicol,2019,38(2):136-140.
[4] YUCELI S,YAZICI G N,MAMMADOV R,et al.The effect of lutein on ischemia-reperfusion-induced vasculitic neuropathic pain and neuropathy in Rats[J].In Vivo,2021,35(3):1537-1543.
[5] RAMEZANIFAR S,BEYRAMI S,MEHRIFAR Y,et al.Occupational exposure to physical and chemical risk factors:a systematic review of reproductive pathophysiological effects in women and men[J].Saf Health at Work,2023,14(1):17-30.
[6] SOLTANPOUR Z,RASOULZADEH Y,MOHAMMADIAN Y.Occupational exposure to metal fumes among iranian welders:Systematic review and simulation-based health risk assessment[J].BTer Medline,2023,201(3):1090-1100.
[7] SAHA S,BUTTARI B,PANIERI E,et al.An overview of Nrf2 signaling pathway and its role in inflammation[J].Molecules,2020,25(22):5474.
[8] FANG Y,GAN C,PENG J,et al.Effects of manganese and iron,alone or in combination,on apoptosis in BV2 cells[J].Biol Trace Elem Res,2023, 9(2):1-12.
[9] MARKIV B,EXPSITO A,RUIZ-AZCONA L,et al.Environmental exposure to manganese and health risk assessment from personal sampling near an industrial source of airborne manganese[J].Env Res,2023, 224(12):115478.
[10] ALIKUNJU M,MISIRIYYAH N,IQBAL S S,et al.Manganese neurotoxicity as a stroke mimic:A case report[J].Cureus,2023,15(4):65-67.
[11] HUAN M A O,YAN L I.Research progress of manganese exposure on abnormal GnRH release in hypothalamus affecting reproductive function[J].J Occup Environ Med,2023,40(1):107-110.
[12] DRAPER M,BESTER M,VAN ROOY M,et al.Adverse pulmonary effects after oral exposure to copper,manganese and mercury,alone and in mixtures,in a Spraque-Dawley rat model[J].Ultrastruct Pathol,2023, 47(3):146-159.
[13] FAVARIN J C,BASOTTI A,BAPTISTELLA A R,et al.Neuroprotective effect of vitamin d on behavioral and oxidative parameters of male and female adult wistar rats exposed to mancozeb(manganese/zinc ethylene bis-dithiocarbamate)[J].Mol Neurobiol,2023,60(7):3724-3740.
[14] WU L,LAN Y,YU Z,et al.Blood manganese and non-alcoholic fatty liver disease in a high manganese exposure area in China[J].J Health Popul Nutr,2023,42(1):118-120.
[15] HORVÁTH G,SZÖKE É,KEMÉNY Á,et al.Lutein inhibits the function of the transient receptor potential A1 ion channel in different in vitro and in vivo models[J].J Mol Neurosci,2012,46(24):1-9.
[16] SHIMAZU Y,KOBAYASHI A,ENDO S,et al.Effect of lutein on the acute inflammation-induced c-Fos expression of rat trigeminal spinal nucleus caudalis and C1 dorsal horn neurons[J].Eur J Oral Sci,2019, 127(5):379-385.
[17] MITRA S,RAUF A,TAREQ A M,et al.Potential health benefits of carotenoid lutein:An updated review[J].Food Chem Toxicol,2021, 154(10):112328.
[18] ULASOV A V,ROSENKRANZ A A,GEORGIEV G P,et al.Nrf2/Keap1/ARE signaling:Towards specific regulation[J].Life Sci,2022, 291(18):120111.
[19] SHAW P,CHATTOPADHYAY A.Nrf2-ARE signaling in cellular protection:Mechanism of action and the regulatory mechanisms[J]. J Cell Physiol,2020,235(4):3119-3130.
[20] YU C,XIAO J H.The Keap1-Nrf2 system:A mediator between oxidative stress and aging[J].Oxid Med Cell Longev,2021,69(2):1-16.