Tan Shuwen, Li Hua, Jin Ying, Yu Hui
College of Life Science, Foshan University, No. 1 Xianhu University Road, Nanhai, Foshan, Guangdong 528231, China; Holdone Aquaculture Breeding Limited Company, Foshan, Guangdong 528231, China.
College of Biophotonics, South China Normal University, Guangzhou, Guangdong 510631, China.
Ecotoxicol Environ Saf. 2014 Oct;108:258-64. doi: 10.1016/j.ecoenv.2014.07.011. Epub 2014 Aug 5.
To gain more knowledge about the physiological regulation of metal pollutant detoxification in grass carp, we examined Cd concentration and its the potential influence on the expression of metallothionein 2 (MT2) and multidrug resistance protein 2 (ABCC2) mRNA in the liver and kidney, using in vitro and in vivo experiments. First, the full-length of MT2 cDNA and partial ABCC2 cDNA was obtained, consisting 183bp and 366bp respectively. In vivo approach, grass carp received 96h exposure of Cd (1/10 LD50), and MT2 and ABCC2 mRNA expression were determined by qRT-PCR. The Cd treatment resulted in an increase of MT2 mRNA level in the liver with Cd accumulation. Nonetheless, the elevation ABCC2 mRNA in the liver was appeared at 48h after Cd exposure, as well as the expression of MT2 and ABCC2 mRNA in the kidney. The in vitro experiment was carried out using the hepatocyte (L86) and nephroblasts (CIK). The qRT-RCR results showed that MT2 and ABCC2 mRNA dramatically increased following Cd exposure (1/10 LD50); however, ABCC2 mRNA expression was suppressed in the L86 cell line at first (6h). In conclusion, this result suggested that both MT2 and ABCC2 mRNA may play important roles in the detoxification of toxic metals, and MT2 gene was more sensitive to Cd induction.
为了更深入了解草鱼体内金属污染物解毒的生理调节机制，我们通过体外和体内实验，检测了镉（Cd）浓度及其对肝脏和肾脏中金属硫蛋白2（MT2）和多药耐药蛋白2（ABCC2）mRNA表达的潜在影响。首先，获得了MT2 cDNA全长和ABCC2 cDNA部分序列，分别为183bp和366bp。在体内实验中，草鱼接受了96小时的Cd（1/10半数致死剂量）暴露，并通过qRT-PCR测定MT2和ABCC2 mRNA表达。Cd处理导致肝脏中MT2 mRNA水平随Cd积累而增加。然而，肝脏中ABCC2 mRNA的升高出现在Cd暴露后48小时，肾脏中MT2和ABCC2 mRNA的表达也是如此。体外实验使用肝细胞（L86）和成纤维细胞（CIK）进行。qRT-RCR结果显示，Cd暴露（1/10半数致死剂量）后MT2和ABCC2 mRNA显著增加；然而，ABCC2 mRNA表达在L86细胞系中最初（6小时）受到抑制。总之，该结果表明MT2和ABCC2 mRNA可能在有毒金属解毒中发挥重要作用，并且MT2基因对Cd诱导更敏感。
