碘过量对金属硫蛋白Ⅰ/Ⅱ敲除小鼠脾细胞的早期影响  

Early effects of iodine excess on spleen cells of methallothionein Ⅰ/Ⅱ knockout mice

作　　者：王凌燕[1] 张娜[1] 李永梅[2] 段琦[1] 姚小梅[1] 

机构地区：[1]天津医科大学病理生理学教研室,300070 [2]天津医科大学内分泌研究所、卫生部激素与发育重点实验室,300070

出　　处：《中华地方病学杂志》2015年第3期168-171,共4页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81273009）

摘　　要：目的 探讨碘过量对金属硫蛋白Ⅰ/Ⅱ敲除（methallothionein Ⅰ/Ⅱknockout,MT-Ⅰ/ⅡKO）小鼠脾细胞活力、乳酸脱氢酶（LDH）活力、脾细胞线粒体超氧化物生成和过氧化物酶（Prx）3表达的影响.方法 选取6～8周龄健康雄性MT-Ⅰ/ⅡKO小鼠和同源非基因敲除（Wildtype,WT）小鼠,取脾组织,制备脾细胞悬液;调整脾细胞个数为5×107个/L,并接种于96孔板中（每孔100 μl）.实验组每孔分别给予10-4、10-3、10-2 mol/L碘化钾（KI）或10-3 mol/L过氧化氢（H2O2）处理2h;对照组不给予KI和H2O2处理.四甲基偶氮唑（MTT）法检测脾细胞活力,化学比色法检测LDH活力,流式细胞术检测脾细胞线粒体超氧化物生成,蛋白质免疫印迹法（Western blot）检测Prx3的蛋白表达.结果 MT-Ⅰ/ⅡKO和WT小鼠各组间脾细胞活力、LDH活力、脾细胞线粒体超氧化物生成和Prx3蛋白表达比较差异均有统计学意义（F=357.92、71.03、130.36、10.36、179.58、26.92、187.43、7.16,P均＜0.05）.其中10-4、10-3、10-2 mol/L KI和10-3 mol/L H2O2组脾细胞活力[（80.77±1.86）％、（89.89±2.90）％,（76.08±1.92）％、（87.66±1.74）％,（73.26±1.86）％、（84.30±2.23）％,（66.22±1.71）％、（70.80±149）％]均低于对照组[（100.00±2D0）％、（100.00±1.63）％,P均＜0.05];LDH活力[（3 880.00±190.62）、（3 431.17±170.45）,（4 178.33±170.43）、（3 598.63±189.09）,（4 388.61±123.79）、（3 863.72±195.64）,（4 615.28±196.17）、（4 148.12±195.81）U/L]均高于对照组[（3 202.22±85.63）、（3 161.51±144.49）U/L,P均＜0.05];线粒体超氧化物生成（53.83±3.22、47.03±1.60,58.92±4.00、50.48±2.59,72.72±2.14、68.53±2.97,80.76±4.11、75.26±3.41）明显高于对照组（43.82±1.56、38.60±2.81,P均＜0.05）;10-3、10-2 mol/L KI和10-3 mol/L H2O2组脾细胞Prx3蛋白表达（0.82±0.12、0.65±0.12,0.96±0.15、0.73±0.16,1.04±0.13、0.85±0.16）均高于对照组（0.61±0.09、0.50±0Objective To investigate the effects of iodine excess on spleen cell viability,lactate dehydrogenase （LDH） leakage,mitochondrial superoxide production and peroxiredoxin （Prx）3 expression in methallothionein Ⅰ / Ⅱ knockout （MT-Ⅰ / Ⅱ KO）mice.Methods Spleen cell suspensions were prepared from six to eight-week old and healthy male MT-Ⅰ / Ⅱ KO mice and wild type （WT） mice; the cell number was adjusted to 5 × 107/L and the cells were plated in 96-well plates （100 μl each well）; the cells were exposed to various concentrations of KI （0,10-4,10-3,10-2 mol/L） and 10-3 mol/L H2O2,respectively,for two hours,and control group did not give KI nor H2O2.Cell viability was assayed by methyl thiazolyl tetrazolium （MTT） colorimetric method.Cell damage was detected by chemical colorimetric method.Mitochondrial superoxide production in the spleen cells was measured by flow cytometry.Western blotting technology was used to investigate the expression of Prx3.Results In both MT-Ⅰ/Ⅱ KO and WT mice,the differences of cell viability,LDH leakage,mitochondrial superoxideproduction and the expression of Prx3 of spleen cells among the treatment groups were statistically significant （F =357.92,71.03,130.36,10.36,179.58,26.92,187.43,and 7.16,all P ＜ 0.05）.Compared to the control group [（100.00 ± 2.00）％,（100.00 ± 1.63）％,（3 202.22 ± 85.63）,（3 161.51 ± 144.49）U/L,43.82 ± 1.56,38.60 ± 2.81,0.61 ± 0.09,0.50 ± 0.08],cell viability of 10-4,10-3,10-2 mol/L KI treatment and 10-3 mol/L H2O2 groups [（80.77 ± 1.86）％,（89.89 ± 2.90）％,（76.08 ± 1.92）％,（87.66 ± 1.74）,（73.26 ± 1.86）％,（84.30 ± 2.23）％,（66.22 ± 1.71）％,（70.80 ± 1.49）％] was decreased （all P ＜ 0.05）; LDH leakage [（3 880.00 ± 190.62）,（3 431.17 ± 170.45）,（4 178.33 ± 170.43）,（3 598.63 ± 189.09）,（4 388.61 ± 123.79）,（3 863.72 ± 195.64）,（4 615.28 ± 196.17）,（4 148.12 ± 195.81）U/L] was increased significantly （all P＜ 0.05）; a

关 键 词：金属硫蛋白 碘 脾 超氧化物类 线粒体 

分 类 号：R599[医药卫生—内科学]