N-乙酰半胱氨酸对氟诱导睾丸支持细胞内质网应激的作用  

Effects of N-acetylcysteine on fluoride-induced endoplasmic reticulum stress in sertoli cells

作　　者：杨阳[1] 黄辉[1] 冯得敏 刘文一[1] 程学敏[1] 巴月[1] 崔留欣[1] 

机构地区：[1]郑州大学公共卫生学院环境卫生学教研室,郑州450001

出　　处：《卫生研究》2014年第5期805-808,813,共5页Journal of Hygiene Research

基　　金：河南省教育厅科学技术研究重点项目(No.13A330735)

摘　　要：目的探讨N-乙酰半胱氨酸对氟化钠(NaF)介导睾丸支持细胞内质网应激的抑制作用。方法将原代培养大鼠睾丸支持细胞进行分组:0μg/ml NaF(对照组)、6μg/ml NaF组、12μg/ml NaF组、24μg/ml NaF组、N-乙酰半胱氨酸(2 mmol/L NAC)组、6μg/ml NaF+2 mmol/L NAC组、12μg/ml NaF+2 mmol/L NAC组和24μg/ml NaF+2 mmol/L NAC组,各组以相应药物孵育24 h。四甲基偶氮唑蓝(MTT)法测定各组细胞生长活性;荧光探针法分析各组细胞内活性氧(ROS)水平;免疫印迹法(Western blot)检测内质网应激相关蛋白GRP78、PERK和CHOP的表达。结果 2 mmol/L NAC有效提高了细胞存活率(P<0.01);NAC明显抑制了NaF介导的细胞内ROS水平的升高,与NaF组相比,各NAC组的ROS水平均显著降低(P<0.01);NaF诱导内质网应激相关蛋白GRP78、PERK和CHOP表达明显上调,经2mmol/L NAC处理后,其有效拮抗了GRP78、PERK和CHOP蛋白表达的上调(P<0.01)。结论 ROS激活了内质网应激相关信号通路,而NAC通过减少ROS的产生,拮抗了NaF介导的睾丸支持细胞内质网应激。Objective Investigated the effects of N-acetylcysteine （NAC） on endoplasmic reticulum stress of sertoli cells induced by sodium fluoride （NaF）. Methods Rat sertoli cells were exposed to various concentration of （0, 6, 12, 24 μg/ml） sodium fluoride with or without 2 mmol/L NAC for 24 hours. The cell viability was evaluated using trypan blue exclusion test. Intracellular reactive oxygen species （ROS） was measured using the fluorescent probe DCFH-DA. Western blot was used to test the expression of GRP78, PERK and CHOP. Results It was found that treatment with NAC（2 mmol/L）restored the reduced cell viability and excessive oxidative stress （P 〈 0.01 ）. Moreover, fluoride exposure upregulated the expression of GRP78,PERK and CHOP protein （P 〈 0. 01 ）. NAC was also found to suppress the levels of GRP78,PERK and CHOP expression in NaF-treated cells （P 〈 0. 01 ）. Conclusion Endoplasmic reticulum stress signaling pathways were activated by ROS, and NAC attenuate endoplasmic reticulum stress through inhibiting thelevels of ROS in NaF-treated sertoli cells.

关 键 词：N-乙酰半胱氨酸 氟化钠 睾丸支持细胞 内质网应激 ROS 

分 类 号：R994.6[医药卫生—毒理学] R595[医药卫生—药学]