Chen Jianjie, Cao Jinling, Luo Yongju, Xie Lingtian, Song Jing, Xue Wenjuan, Jia Ruhui, Song Jie
State Key Laboratory of Ecological Animal Husbandry and Environmental Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi, PR China.
Guangxi Academy of Fishery Sciences, Nanning 530021, Guangxi, PR China.
Acta Histochem. 2014 Oct;116(8):1337-41. doi: 10.1016/j.acthis.2014.08.005. Epub 2014 Sep 2.
Chronic exposure to fluoride can result in a variety of adverse effects in fish. Previously we indicated that high fluoride caused damage and apoptosis in the kidneys of the common carp, Cyprinus carpio. In this study, the effects of fluoride on the expression and localization of ERK and p-ERK proteins in the ERK signaling pathway were determined using Western blotting and immunohistochemical methods in the kidneys of carp exposed to 0, 40, 80, 120mg/L fluoride, respectively. Western blotting analysis found that compared with the controls, the levels of ERK1 and ERK2 proteins were relatively unchanged in fluoride-exposed fish, while p-ERK1 and p-ERK2 protein levels decreased significantly with the increased fluoride concentrations. The immunohistochemical analysis found the proteins of ERK and p-ERK were predominantly localized in the cytoplasm of epithelial cells in the renal tubules of C. carpio. Compared with the control group, the levels of ERK protein were relatively constant, yet the levels of p-ERK protein and p-ERK/ERK ratio were reduced with fluoride exposure dose. These findings indicate that the renal damage in carp exposed to fluoride is mediated via the ERK pathway. Fluoride exposure could inactivate ERK, inhibit the expression of p-ERK protein, and induce renal damage in C. carpio.
长期接触氟化物会对鱼类产生多种不良影响。此前我们指出,高氟会导致鲤鱼(Cyprinus carpio)肾脏损伤和细胞凋亡。在本研究中,分别采用蛋白质免疫印迹法和免疫组织化学方法,检测了暴露于0、40、80、120mg/L氟化物的鲤鱼肾脏中,氟化物对ERK信号通路中ERK和p-ERK蛋白表达及定位的影响。蛋白质免疫印迹分析发现,与对照组相比,氟暴露组鱼类中ERK1和ERK2蛋白水平相对未变,而p-ERK1和p-ERK2蛋白水平随氟化物浓度升高而显著降低。免疫组织化学分析发现,ERK和p-ERK蛋白主要定位于鲤鱼肾小管上皮细胞的细胞质中。与对照组相比,ERK蛋白水平相对恒定,但p-ERK蛋白水平和p-ERK/ERK比值随氟暴露剂量增加而降低。这些结果表明,氟暴露导致的鲤鱼肾脏损伤是通过ERK信号通路介导的。氟暴露可使ERK失活,抑制p-ERK蛋白表达,并诱导鲤鱼肾脏损伤。