1. 大连海洋大学 辽宁省海洋生物资源恢复与生境修复重点实验室, 辽宁 大连 116023;
2. 大连海洋大学 农业部北方海水增养殖重点实验室, 辽宁 大连 116023
吴迪(1991-),女,硕士,专业方向为海洋生物学.E-mail:15842623565@163.com
X171
国家自然科学基金项目（31272650）.
1. Key Laboratory of Marine Bio-resource Restoration and Habitat Reparation in Liaoning Province, Dalian Ocean University, Dalian 116023, China;
2. Key Laboratory of Mariculture, Ministry of Agriculture;Dalian Ocean University, Dalian 116023, China
以二倍体和三倍体泥鳅（）鳍细胞系（DIMF和TRMF）为实验材料，研究了重铬酸钾对细胞的氧化损伤、微核形成以及金属硫蛋白表达情况的影响，旨在多角度探讨重铬酸钾对细胞系的毒性效应，建立适合监测其污染情况的指标。采用MTT法测定了细胞的半数抑制浓度；使用试剂盒测定了3种主要抗氧化酶活性；吉姆萨染色后观察了细胞微核的变化，并通过实时定量PCR方法测定了金属硫蛋白的表达情况。结果表明，24 h急性毒性实验两种细胞系的半抑制浓度分别为（25.3±1.2）μmol/L、（27.9±0.6）μmol/L，DIMF对重铬酸钾的敏感性要高于TRMF。当重铬酸钾浓度为0~30 μmol/L时，二个细胞系的超氧化物歧化酶（SOD）活性随染毒浓度升高而升高。当重铬酸钾浓度为0~20 μmol/L时，谷胱甘肽过氧化物酶（GSH-Px）活性逐渐升高，浓度为30~40 μmol/L时，其活力开始下降。两种细胞系的谷胱甘肽S转移酶（GST）活性随重铬酸钾浓度增大逐渐降低。二倍体细胞系氧化酶活性均低于三倍体。微核试验显示，重铬酸钾可引起细胞核损伤，形成微核。当重铬酸钾浓度为40 μmol/L时，DIMF、TRMF微核率达到最大，分别为7.33‰和8.00‰，DIMF微核率要低于TRMF。实时定量PCR结果显示，对照组金属硫蛋白（<0.01）。
Waste water discharge containing chromium can lead to chromium polluted regional water. Because of the accumulation and enrichment of chromium, fish and other aquatic organisms can be injured seriously in polluted water, and chromium can cause toxic effects or death in humans if ingested. Chromium mainly refers to Cr6+, which exists as a noncatalytic form or as potassium dichromate. Fish, such as loach and carp, are commonly used to detect chromium, but few reports have used fish cell lines. Previous studies have revealed that cells cultured are homogeneous, respond quickly, and are convenient to use. In this study, loach fin cell lines established from diploids and triploids (DIMF and TRMF) were used to assess the toxic effects of potassium dichromate . The purpose of this study was to establish suitable indicators to monitor chromium pollution. The concentration that inhibited 50% of the cells (IC50) was determined using the thiazolyl blue (MTT) method, the activities of three main antioxidants were tested with kits, the changes in cell nuclei were observed after Giemsa staining, and expression of the metallothionein () gene was measured by real-time polymerase chain reaction (PCR) analysis. The results showed that the 24-h IC50 values of DIMF and TRMF were (25.3±1.2) mol/L and (27.9±0.6) μmol/L, respectively; the sensitivity of DIMF to potassium dichromate was higher than that of TRMF, and the sensitivities of the two cell lines were higher than those of loach in vivo. Superoxide dismutase (SOD) activities in the two cell lines increased as the potassium dichromate concentration was increased from 0 to 30 μmol/L. Glutathione peroxidase (GSH-Px) activities increased in the two cell lines as the potassium dichromate concentration was increased from 0 to 20 μmol/L; however, GSH-Px activities decreased when concentration was 30-40 μmol/L. Glutathione S-transferase (GST) activities in the two cell lines decreased gradually as the potassium dichromate concentration was increased. Overall, the enzymatic activities of DIMF were lower than those of TRMF. Potassium dichromate caused nuclear damage and micronuclei formed. The maximum rates of micronuclear formation in DIMF and TRMF were 7.33‰ and 8.00‰, respectively when the potassium dichromate concentration was 40 μmol/L. The rate of micronuclear formation was lower in DIMF than that in TRMF. The real-time PCR results showed that MT gene expression in the control group was very low, but increased significantly in response to potassium dichromate stress (<0.01). MT gene expression was maximal in response to 30 μmol/L potassium dichromate and was 49.9-and 50.7-times higher than that of the control group, respectively. However, the difference between DIMF and TRMF was not significant.
吴迪,李霞,秦艳杰,白丽雯,周诗珈.重铬酸钾对不同倍性泥鳅鳍细胞系的毒性效应[J].中国水产科学,2017,24(1):173-179
WU Di, LI Xia, QIN Yanjie, BAI Liwen, ZHOU Shijia. Toxic effects of potassium dichromate on loach fin cell lines with different ploidies in vitro[J]. Journal of Fishery Sciences of China,2017,24(1):173-179
