不同水平钙对氟中毒大鼠肾组织细胞内质网凋亡通路的影响

doi:10.11843/j.issn.0366-6964.2018.12.022

摘要/Abstract

摘要：

为探讨不同水平钙对氟中毒大鼠肾组织细胞损伤内质网凋亡通路的影响，将120只雄性SD大鼠随机分为对照组（C）（常规饲料）、高氟组（F）（常规饲料，含150 mg·kg^-1 F^-）、高氟低钙组（L）（常规饲料，含150 mg·kg^-1 F^-+0.5% CaCO₃）、高氟中钙组（M）（常规饲料，含150 mg·kg^-1 F^-+1% CaCO₃）和高氟高钙组（H）（常规饲料，含150 mg·kg^-1 F^-+2% CaCO₃），自由采食和自由饮水120 d，试验结束后取肾组织进行病理学检测，分别运用RT-PCR、免疫组织化学法和Western blot技术检测肾组织细胞中内质网通路相关基因及蛋白的表达。结果显示：（1）氟中毒可使大鼠肾小球肿胀，肾球囊间隙变宽，近曲小管上皮细胞肿胀并出现空泡变性，低、中剂量的钙可使氟的肾毒性减轻，而高剂量钙组损伤更加严重；（2）高氟组内质网凋亡通路中Caspase-12、Caspase-3和JNK基因mRNA水平表达显著升高，IRE1、ASK1、TRAF2基因表达显著下降，但L组和M组中其表达量有不同程度的改变，缓解了氟的肾毒性；（3）高氟组内质网通路Bcl-2蛋白表达量显著下降，Caspase-12、JNK、Bax蛋白及Bax/Bcl-2的比值显著升高，而补充低、中剂量的钙可缓解内质网通路相关蛋白的过表达，从而抑制氟的毒性作用。高氟可诱发肾组织细胞内质网通路中Caspase-12信号通路和JNK信号通路导致肾细胞凋亡加速，而低、中剂量的钙可通过抑制肾组织细胞中内质网凋亡通路相关基因及蛋白的表达缓解氟的毒性作用。

Abstract:

This study was conducted to investigate the effects of different levels of calcium on the endoplasmic reticulum apoptosis pathway in fluoride-induced kidney tissue injury in rats. One hundred and twenty male SD rats were randomly divided into 5 groups:Control group (C) (conventional feed), High fluoride group (F) (conventional feed, containing 150 mg·kg^-1 F^-), High fluoride + Low calcium (L) (conventional feed containing 150 mg·kg^-1 F^-+0.5% CaCO₃), High fluoride + Medium calcium group (M) (conventional feed containing 150 mg·kg^-1 F^-+1% CaCO₃) and High fluoride + High calcium group (H) (conventional feed containing 150 mg·kg^-1 F^-+2% CaCO₃). After eating and drinking freely for 120 days, renal tissue cells were collected for pathological examination. The expression of endoplasmic reticulum pathway related genes and proteins were detected by RT-PCR, immunohistochemistry and western blot, respectively. Results showed that:(1) Fluorosis caused glomerular swelling, widening of the renal balloon space, swelling of proximal tubule epithelial cells and vacuole degeneration in rats. Low and medium doses of calcium reduced fluoride-induced kidney toxicity, while the high-dose calcium group enhanced the injury. (2) Fluoride significantly increased the mRNA expression of Caspase-12, Caspase-3 and JNK, while the expression of IRE1, ASK1 and TRAF2 was significantly decreased, which are endoplasmic reticulum apoptosis pathway related genes. However, low and medium doses of calcium relieved the fluoride-induced adverse effects. (3) In addition, fluoride significantly decreased the protein expression of Bcl-2 and increased the protein levels of Caspase-12, JNK, Bax, Bax/Bcl-2. Supplementation of low and medium doses of calcium alleviated the over-expression of endoplasmic reticulum pathway related proteins, thereby inhibiting the toxic effects of fluoride. In summary, fluoride activated the Caspase-12 signaling pathway and the JNK signaling pathway of endoplasmic reticulum pathway, which accelerated the apoptosis of renal tissue cells. However, low and medium doses of calcium relieved the toxic effects of fluoride by inhibiting the expressions of endoplasmic reticulum apoptosis pathway related genes and proteins.

中图分类号:

王金明, 徐慧淼, 张杰, 高宇凤, 赵阳飞, 李妍妍. 不同水平钙对氟中毒大鼠肾组织细胞内质网凋亡通路的影响[J]. 畜牧兽医学报, 2018, 49(12): 2723-2732.

WANG Jin-ming, XU Hui-miao, ZHANG Jie, GAO Yu-feng, ZHAO Yang-fei, LI Yan-yan. Effects of Different Calcium Levels on the Apoptosis Pathways of Renal Tissue Cells in Rats with Fluorosis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2723-2732.

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