Mi Yuling, Zhang Caiqiao
Department of Veterinary Medicine, College of Animal Sciences, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, China.
Toxicol Lett. 2005 Feb 15;155(2):297-305. doi: 10.1016/j.toxlet.2004.10.005.
The toxic and hormonal effects of polychlorinated biphenyls (PCBs) on testicular germ cell development were revealed with a germ-Sertoli cell co-culture model from embryonic chickens. Testicular cells were dispersed from 18-day-old embryo and exposed to Aroclor 1254 (A1254) alone and combined with alpha-tocopherol, flutamide and tamoxifen for discretion of the toxic and hormonal actions of A1254. Cell damage was evaluated by determinations of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction and lactate dehydrogenase (LDH) leakage. Results showed that 10 microg/ml A1254 induced condensed nuclear and vacuolated cytoplasm; cell exfoliated and broke into pieces as a sign of cell degeneration after treatment for 6 h. The morphological cytotoxicity was confirmed by MTT reduction and LDH leakage assays. alpha-Tocopherol attenuated the toxic effect of A1254. After culture for 48 h, A1254 (0.1-1 microg/ml) manifested obvious hormonal effect to induce germ cell proliferation, while 10 microg/ml A1254 displayed obvious toxic and hormonal effects on germ and somatic cells. Blocking of either androgen receptor by flutamide or estrogen receptor by tamoxifen inhibited hormonal effect of A1254 on germ cell proliferation and increased the cytotoxicity. The above results indicated that A1254 exposure imposed both toxic and hormonal effects on embryonic testicular germ cell proliferation, which may cause reproductive disorder and even infertility at adulthood.
利用来自胚胎鸡的生殖细胞-支持细胞共培养模型,揭示了多氯联苯(PCBs)对睾丸生殖细胞发育的毒性和激素作用。从18日龄胚胎中分离出睾丸细胞,使其单独暴露于Aroclor 1254(A1254)以及与α-生育酚、氟他胺和他莫昔芬联合暴露,以判断A1254的毒性和激素作用。通过测定3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑(MTT)还原率和乳酸脱氢酶(LDH)泄漏量来评估细胞损伤。结果显示,10微克/毫升的A1254可诱导细胞核浓缩和细胞质空泡化;处理6小时后,细胞脱落并破碎成碎片,作为细胞变性的标志。MTT还原率和LDH泄漏量测定证实了形态学上的细胞毒性。α-生育酚减弱了A1254的毒性作用。培养48小时后,A1254(0.1 - 1微克/毫升)表现出明显的激素作用,可诱导生殖细胞增殖,而10微克/毫升的A1254对生殖细胞和体细胞均表现出明显的毒性和激素作用。氟他胺阻断雄激素受体或他莫昔芬阻断雌激素受体均抑制了A1254对生殖细胞增殖的激素作用,并增加了细胞毒性。上述结果表明,暴露于A1254对胚胎睾丸生殖细胞增殖具有毒性和激素作用,这可能导致成年期的生殖障碍甚至不育。