摘要
[背景]孕鼠补充N-乙酰半胱氨酸(NAC)可缓解镉所致胎鼠生长受限,但是孕期补充NAC对镉致其子代肾脏发育的影响尚不清楚。[目的]本研究旨在探究孕鼠补充NAC对镉暴露致其雄性子代肾脏发育损伤的影响及其相关机制。[方法]65只CD-1孕鼠饲养至妊娠第7天(GD7)后,随机分为对照组(12只)、NAC组(13只)、低浓度镉组(LCd组,12只)、高浓度镉组(HCd组,13只)和NAC加高浓度镉组(NCd组,15只)。LCd组、HCd组和NCd组孕鼠从GD8—GD17经饮水暴露不同质量浓度氯化镉溶液(50、150、150 mg·L^(-1)),NAC组和NCd组孕鼠从GD7—GD17每日给予NAC灌胃一次(500 mg·kg^(-1))。每组4~5只GD18孕鼠被颈椎脱臼处死并获取母鼠血清、胎鼠肾脏;剩余孕鼠自然分娩,其雄性仔鼠在出生后第35天(PND 35)和PND 98颈椎脱臼处死并获取仔鼠血清和肾脏,检测仔鼠肾功能,观察肾脏组织病理结构,并用Image J 1.51软件计算鲍曼囊腔面积,用Western blotting方法检测胎鼠肾脏烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)、超氧化物歧化酶2(SOD2)和不同发育阶段仔鼠肾脏增殖细胞核抗原(PCNA)蛋白表达水平。[结果]孕期母体低浓度镉暴露明显降低胎儿期子代肾脏质量、肾脏系数和青春期子代肾脏质量(均P<0.05)。孕期母体高浓度镉暴露降低胎儿期、青春期子代肾脏质量和肾脏系数及成年期子代肾脏质量(P<0.05)。孕期母体低浓度镉暴露和高浓度镉暴露抑制子代肾脏增殖相关蛋白PCNA表达(均P<0.05);LCd、HCd组青春期和成年期仔鼠肾脏鲍曼囊腔面积均较对照组减小[青春期:(4346.45±205.27)、(4516.20±113.44)μm^(2)vs(17823.50±150.55)μm^(2);成年期:(4774.00±843.29)、(4483.97±146.47)μm^(2)vs(17869.81±665.93)μm^(2);均P<0.01]。与对照组相比,孕期镉暴露上调胎鼠肾脏氧化应激相关蛋白NOX4和SOD2表达水平(均P<0.05);而孕期母体NAC补充明显缓解高浓度镉所致胎肾氧化应激(P<0.05),逆转高浓度镉对仔鼠肾脏PCNA蛋白表达的下调效应(P<0.05),拮抗高浓度镉降低青春期和成年期仔鼠肾脏质量[(0.56±0.02)g vs(0.43±0.01)g,(0.66±0.02)g vs(0.58±0.03)g;均P<0.05]和鲍曼囊腔面积[(21158.41±971.19)μm^(2)vs(4516.20±113.44)μm^(2),(19538.48±178.13)μm^(2)vs(4483.97±146.47)μm^(2);均P<0.01]。[结论]孕期母体补充NAC明显缓解镉所致其雄性子代肾脏发育异常,与拮抗胎鼠肾脏氧化应激反应有关。
[Background]Previous studies have found that maternal N-acetyl-cysteine(NAC)supplementation during pregnancy can alleviate cadmium-induced fetal growth restriction in mice.However,the effect on cadmium-induced renal development impairment in offspring is unclear.[Objective]The present study is to explore the effect of maternal NAC supplementation during pregnancy on Cd-impaired renal development in male mouse offspring and its mechanisms.[Methods]A total of 65 CD-1 pregnant mice were raised to gestational day7(GD7)and randomly divided into control group(n=12),NAC group(n=13),low-concentration cadmium(LCd)group(n=12),high-concentration cadmium(HCd)group(n=13),and NAC plus highconcentration cadmium(NCd)group(n=15).In the LCd group,HCd group,and NCd group,pregnant mice were exposed to different concentrations of cadmium(50,150,and 150 mg·L^(-1))by drinking water from GD8 to GD17.In the NAC group and NCd group,pregnant mice were treated with NAC(500 mg·kg^(-1))daily by intragastrical administration from GD7 to GD17.Four to five pregnant mice in each group were sacrificed by cervical dislocation to collect maternal serum and fetal kidneys on GD18.The remaining pregnant mice delivered naturally;and the male offspring mice were raised to postnatal day 35(PND 35)and PND 98,and sacrificed by cervical dislocation to collect their serum and kidneys.The levels of renal function indicators in male offspring mice serum were detected.Kidney pathological changes were observed.Bowman space area was measured by Image J 1.51.The expression levels of nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4),super oxygen dehydrogenises 2(SOD2)in fetal kidney and proliferating cell nuclear antigen(PCNA)in kidney of offspring mice at different developmental stages were detected by Western blotting.[Results]The maternal low-concentration cadmium exposure during pregnancy reduced the weight and coefficient of fetal kidneys and the weight of adolescent kidneys(P<0.05).The maternal high-concentration cadmium exposure during pregnancy reduced the weight and coefficient of fetal and adolescent kidneys and the weight of adulthood kidneys(P<0.05).The maternal low-concentration and high-concentration cadmium exposure during pregnancy down-regulated the expression of PCNA,a proliferation-associated protein,in offspring kidneys(P<0.05).The LCd group showed decreased Bowman space area in adolescent and adulthood kidneys compared with the control group[(4346.45±205.27)μm^(2)vs(17823.50±150.55)μm^(2),(4774.00±843.29)μm^(2)vs(17869.81±665.93)μm^(2);P<0.01].So did the HCd group[(4516.20±113.44)μm^(2)vs(17823.50±150.55)μm^(2),(4483.97±146.47)μm^(2)vs(17869.81±665.93)μm^(2);P<0.01].The maternal cadmium exposure during pregnancy increased the levels of NOX4 and SOD2 in fetal kidneys(P<0.05).However,the maternal NAC supplementation during pregnancy markedly alleviated the oxidative stress in fetal kidneys(P<0.05),reversed the down-regulated PCNA levels in offspring kidneys(P<0.05),and antagonized the decreases of kidney weight[(0.56±0.02)g vs(0.43±0.01)g,(0.66±0.02)g vs(0.58±0.03)g;P<0.05]and Bowman space area[(21158.41±971.19)μm^(2)vs(4516.20±113.44)μm^(2),(19538.48±178.13)μm^(2)vs(4483.97±146.47)μm^(2);P<0.01]in adolescent and adult offspring kidneys induced by the high-concentration cadmium exposure.[Conclusion]Maternal NAC supplementation during pregnancy could markedly alleviate Cd-impaired renal development in male mouse offspring,which is related to the antagonism of oxidative stress in fetal kidneys.
作者
曹学林
朱华龙
熊永伟
刘韦伯
易颂佳
时雪婷
周国祥
戴丽敏
王华
CAO Xuelin;ZHU Hualong;XIONG Yongwei;LIU Weibo;YI Songjia;SHI Xueting;ZHOU Guoxiang;DAI Limin;WANG Hua(Department of Toxicology,Environmental Toxicology Laboratory of Anhui Higher Education Institutes,Anhui Medical University,Hefei,Anhui 230032,China;School of Public Health,Environmental Toxicology Laboratory of Anhui Higher Education Institutes,Anhui Medical University,Hefei,Anhui 230032,China)
出处
《环境与职业医学》
CAS
CSCD
北大核心
2021年第5期473-481,共9页
Journal of Environmental and Occupational Medicine
基金
国家自然科学基金(81973079)
2020年安徽省自然科学基金杰青(2008085J38)。
关键词
镉
N-乙酰半胱氨酸
孕期
肾脏
氧化应激
增殖
cadmium
N-acetyl-cysteine
pregnancy
kidney
oxidative stress
proliferation
