摘要
目的:流行病学调查结果显示,地方性氟中毒与甲状腺肿的流行有很大的重叠性。探讨氟化钠对原代培养猪甲状腺细胞及甲状腺过氧化物酶活性的影响。方法:实验于2006-10/2007-05在辽宁医学院科学实验室中心完成。①实验方法:在猪死亡1h内取其甲状腺组织,去除甲状腺组织包膜及外层活性差的组织,选取中央活性高的部位,剪成1mm3组织块,用胰蛋白酶、胶原酶Ⅳ消化分离,得到含猪甲状腺细胞的消化液,沉淀悬于含体积分数为0.15的胎牛血清、1U/L牛促甲状腺素、80万U/L庆大霉素的F12培养基中,过滤,调整细胞浓度至5×105L-1,接种培养,每3d更换培养液。常规培养48h的猪甲状腺细胞接种于96孔培养板,按氟化钠终浓度不同分为0,40,80,160mg/L组。②实验评估:染毒48h后,采用噻唑蓝法测定细胞存活量,采用改良愈创木酚法测定甲状腺过氧化物酶活性。结果:①氟化钠对原代培养猪甲状腺细胞存活量的影响:与0mg/L氟化钠比较,40mg/L氟化钠染毒后猪甲状腺细胞的存活量基本相似;80,160mg/L氟化钠染毒后猪甲状腺细胞的存活量均明显下降(P<0.01)。②氟化钠对甲状腺过氧化物酶活性的影响:与0mg/L氟化钠比较,40,80,160mg/L氟化钠染毒后的甲状腺过氧化物酶活性均明显下降(P<0.01),呈剂量-效应关系。结论:氟化钠对原代培养的猪甲状腺细胞及甲状腺过氧化物酶活性均具有抑制作用。
AIM： Endemic fluorosis and the prevalence of goiter have a great overlap by some epidemiologicel investigations. This article is aimed to approach the effect of sodium fluoride （NaF） on the primary porcine thyroid cells and thyroid peroxidase activity. METHODS： The experiment was performed in the Center of Scientific Experiment, Liaoning Medical University from October 2006 to May 2007. ①The thyroid tissues were cut in 1 hour after the pigs had died. The outer envelope and poor activity tissues were removed, and these ones with high activity in the center were selected. The tissues were cut into 1 mm^3 and digested by trypsinase and collagenase Ⅳ. Digestive juice containing porcine thyroid cells was gain. These cells were suspended in F-12 medium containing fetal bovine serum of 0.15 volume fraction, 1 U/L bull thyrotropic-stimulating hormone （TSH） and 800 thousands U/L gentamycin, filtered and inoculated culture by 5×10^5· L^-1. The nutrient solution was replaced every three days. Thyroid cells cultured for 48 hours were inoculated in 96-well culture plate. The experiment was divided into 4 groups according to the final concentration of NaF, 0 mg/L, 40 mg/L, 80 mg/L and 160 mg/L injured concentration groups. ②The cell survival quantity was measured with MTT method and the thyroid peroxidase activity was measured with modified guaiacol method 48 hours later. RESULTS： ①There was no significant difference in cell survival quantity between the 40 mg/L NaF injured group and the 0 mg/L NaF group. Thyroid cells survival quantity was significantly lower in the 80, 160 mg/L NaF injured groups than that in the 0 mg/L NaF group （P 〈 0.01 ）. ②The thyroid peroxidase activity was significantly decreased in the 40, 80, 160 mg/L NaF injured groups than the 0 mg/L NaF group （P〈 0.01 ） and displayed dose-related effect. CONCLUSION： NaF has a significant inhibitory action on the primary porcine thyroid cells and thyroid peroxidase activity.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第37期7425-7428,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
