氟与砷单独及联合暴露对子代大鼠肾脏毒性损伤与细胞自噬的影响

doi:10.3969/j.issn.1004-616x.2022.03.002

摘要/Abstract

摘要： 目的: 评估妊娠期至成年期大鼠慢性氟与砷暴露对肾脏的毒性损伤,并探究潜在的调控机制,为完善氟与砷中毒的发病机制提供科学依据。方法: 试验设置4组即对照组(蒸馏水)、氟暴露组(100 mg/L氟化钠)、砷暴露组(75 mg/L亚砷酸钠)、氟与砷联合暴露组(100 mg/L氟化钠+75 mg/L亚砷酸钠),暴露对象为子代大鼠,从孕鼠妊娠第1天到出生后第21天分别通过母体子宫与母乳的方式暴露于上述氟、砷毒性环境;断乳后,通过饮水染毒至120 d。取子代雄性大鼠新鲜肾脏组织,称取质量后计算肾系数,将肾脏皮质部分通过Masson染色及透射电镜观察肾小球与肾小管的病理改变,并采用免疫荧光技术测定肾小球与肾小管中细胞自噬相关蛋白的表达强度及分布。结果: 与对照组相比,砷暴露组肾系数显著增加(P<0.01);氟、砷单独及联合暴露后肾脏纤维化程度加重,其中氟暴露组肾脏纤维化最明显;此外,所有暴露组中肾小球系膜细胞增生,足细胞内部线粒体空泡化,其中在砷暴露组足细胞中发现典型的自噬小体;相较于对照组,氟、砷单独及联合暴露后肾脏组织中自噬相关蛋白Beclin1、LC3、p62表达水平显著增加(P<0.05),且肾小球与肾小管中自噬相关蛋白的荧光强度与荧光面积也明显增加(P<0.01)。结论: 妊娠期至成年期大鼠氟与砷单独及联合暴露引起肾脏组织中肾小球与肾小管的毒性损伤,并通过增加自噬相关蛋白Beclin1、LC3、p62的表达而调控肾脏细胞自噬的发生。

关键词: 氟暴露, 砷暴露, 氟与砷联合暴露, 肾损伤, 细胞自噬

Abstract: OBJECTIVE: To evaluate toxicity from chronic fluoride and arsenic exposures to kidneys during pregnancy through adulthood in rats,and to explore the potential regulatory mechanisms. METHODS: Rats were divided into four groups:control (distilled water),fluoride exposure (100 mg/L sodium fluoride),arsenic exposure (75 mg/L sodium arsenate),and fluoride and arsenic combined exposure (100 mg/L sodium fluoride+75 mg/L sodium arsenic) groups.The methods of exposure were as follows:from the first day of pregnancy to the 21st day after birth,offspring rats were exposed to fluoride and arsenic toxicity through maternal uterus and breast milk respectively.After weaning,exposure through free drinking water up to 120 days through the same method of exposure as the parents.The kidney coefficient was calculated after weighing the fresh kidney tissues of the offspring male rats.Pathological changes of glomerulus and tubules were observed by Masson staining and transmission electron microscopy.Expression intensity and distribution of autophagy-related proteins in glomerulus and tubules were determined by immunofluorescence technique. RESULTS: Compared with the control group,the renal organ coefficient of the arsenic exposure group was significantly increased (P<0.01).The degrees of renal fibrosis were aggravated after alone and combined exposure of fluoride and arsenic,and renal fibrosis was the most obvious in the fluoride exposed group.Hyperplasia of mesangial cells and vacuolation of mitochondria in podocytes were found in all exposed group.Typical autophagosomes in podocytes were found in the arsenic exposed group.Compared with the control group,the expression levels of autophagy related proteins Beclin1,LC3 and p62 in renal tissues were significantly increased after alone and combined exposure of fluoride and arsenic (P<0.05).Fluorescence intensity and fluorescence area of autophagy-related proteins of glomerulus and renal tubules were also markedly increased (P<0.01). CONCLUSION: Alone and combined exposures to fluoride and arsenic from pregnancy to adulthood induced toxic damage to glomerulus and tubules in renal tissues.Moreover,autophagy of kidney was associated with increasing expressions of autophagy related proteins Beclin1,LC3 and p62.

Key words: fluoride exposure, arsenic exposure, combined exposure of fluoride and arsenic, kidney injury, autophagy

中图分类号:

R994.6

田晓琳, 杨玲玲, 赵倩, 仇玉兰, 孙子龙, 牛瑞燕, 阎小艳. 氟与砷单独及联合暴露对子代大鼠肾脏毒性损伤与细胞自噬的影响[J]. 癌变·畸变·突变, 2022, 34(3): 169-177.

TIAN Xiaolin, YANG Lingling, ZHAO Qian, CHOU Yulan, SUN Zilong, NIU Ruiyan, YAN Xiaoyan. Effects of fluoride and arsenic alone and combined exposures on nephrotoxicity and autophagy in offspring rats[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(3): 169-177.

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