T-2毒素对亲代母鼠及子代小鼠的毒性作用研究

Toxicity test of T-2 toxin on parental mice and their offspring

目的探讨不同剂量T-2毒素对亲代及子代小鼠血清细胞因子表达及关节软骨的病理学改变的影响。方法健康SPF级CD-1小鼠雌性100只，雄性25只，适应饲养1周。定期交配，发现阴道栓的24 h作为怀孕的第0天，实验动物即为孕鼠。将孕鼠按体重采用随机数字表法分为高、中、低剂量组和对照组（饲料：T-2毒素剂量分别为1 200、600、300、0 μg/kg），每组16-18只。高、中、低剂量组自怀孕第0天开始食用染毒饲料，对照组食用标准饲料，亲代孕鼠自然分娩后其子代继续按原分组染毒，直至子代小鼠成年。检测并记录亲代孕中期母鼠及断乳后母鼠，子代断乳后小鼠及成年后小鼠的血清学指标白细胞介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α、脏器系数及关节软骨病理学变化。结果对照组和低、中、高剂量组亲代孕中期母鼠IL-1β、IL-6及TNF-α含量[（219.56 ± 19.32）、（136.89 ± 20.41）、（210.49 ± 21.23）、（207.41 ± 21.23），（192.73 ± 22.43）、（136.25 ± 29.55）、（187.43 ± 39.32）、（232.48 ± 39.32），（1 303.02 ± 142.10）、（1 072.60 ± 78.30）、（1 065.03 ± 37.44）、（1 169.72 ± 104.18）ng/L]组间比较差异有统计学意义（F = 17.124、6.237、7.670，P均〈 0.05）。低剂量组IL-1β、IL-6含量明显低于中、高剂量组及对照组（P均〈 0.05），低、中、高剂量组TNF-α含量均低于对照组（P均〈 0.05）。对照组和低、中、高剂量组子代断乳小鼠IL-1β、IL-6及TNF-α含量[（142.36 ± 13.36）、（113.01 ± 8.65）、（102.13 ± 8.31）、（123.42 ± 10.41），（109.92 ± 9.76）、（100.26 ± 15.60）、（85.25 ± 9.97）、（100.21 ± 16.46），（1 308.45 ± 204.90）、（1 248.60 ± 96.85）、（1 081.09 ± 105.51）、（1 204.87 ± 153.96）ng/L]组间比较差异有统计学意义（F = 49.823、10.530、7.490，P均〈 0.05）。对照组IL-1β、IL-6含量明显高于低、中及高剂量组（P均〈 0.05），对照组TNF-α含量明显高于中、高剂量组（P均〈 0.05）。子代成年小鼠IL-1β、IL-6及TNF-α含量[（69.71 ± 9.61）、（61.31 ± 10.07）、（63.07 ± 10.39）、（58.56 ± 9.69），（172.55 ± 24.55）、（146.91 ± 13.47）、（151.02 ± 24.93）、（157.21 ± 17.86），（1 136.87 ± 137.39）、（1 002.22 ± 86.52）、（987.12 ± 130.80）、（1 047.21 ± 171.64）ng/L]组间比较差异有统计学意义（F = 4.670、5.636、4.775，P均〈 0.05），3种细胞因子含量染毒组均明显低于对照组（P均〈 0.05）。各组亲代孕中期母鼠胸腺、脾脏及肝脏的脏器系数[（0.14 ± 0.03）、（0.20 ± 0.06）、（0.15 ± 0.02）、（0.12 ± 0.03），（0.71 ± 0.16）、（0.78 ± 0.14）、（0.77 ± 0.15）、（0.38 ± 0.10），（6.19 ± 0.43）、（5.57 ± 0.57）、（6.04 ± 0.32）、（5.11 ± 0.29）]组间比较差异有统计学意义（F = 4.056、11.064、8.312，P均〈 0.05），胸腺系数低剂量组明显高于对照组（P 〈 0.05），脾脏系数高剂量组明显低于对照组（P 〈 0.05），肝脏系数低剂量组及高剂量组均明显低于对照组（P均〈 0.05）。子代断乳小鼠脑系数[（3.45 ± 0.73）、（3.11 ± 0.31）、（2.98 ± 0.45）、（3.04 ± 0.22）]出现明显变化（F = 7.529，P 〈 0.05），染毒组明显下降（P均〈 0.05）。亲代孕中期母鼠及断乳母鼠骺板软骨出现不同程度的异常变化，孕中期随着染毒剂量的增大骺板软骨异常越严重，断乳母鼠随着染毒剂量的增大骺板软骨异常也越严重，且断乳母鼠骺板软骨异常更为严重。结论T-2毒素暴露可造成孕鼠及子代小鼠血中IL-1β、IL-6及TNF-α细胞因子水平下降，还可引起小鼠软骨损伤，且骨损伤程度随着染毒剂量的增加及染毒时间的延长而加重。

ObjectiveTo investigate the effects of different doses of T-2 toxin on the expression of cytokines cytokines and pathological changes in parental mice and their offspring.MethodsOne hundred female mice and 25 male mice （CD-1, SPF） were adapted for one week. After regular random mating, observation of vaginal suppository within the first 24 hours was as the 0th day of pregnancy. The pregnant rats were divided into high dose, medium dose, low dose and control groups according to body weight by a random number table （Feed： the doses of T-2 toxin were 1 200, 600, 300, and 0 μg/kg, respectively）, with 16-18 rats in each group. The high, middle and low dose groups began to consume the poisoned feed on the 0th day of pregnancy, while the control group consumed the standard feed. After natural delivery, their offspring were continually treated the same way as their mother until the offspring reached adulthood. Serum levels of interleukin-1β （IL-1β） and interleukin-6 （IL-6）, tumor necrosis factor-α （TNF-α）, organ coefficient and pathological changes of articular cartilage were determined.ResultsThe levels of IL-1β, IL-6 and TNF-α in the control, low, middle and high T-2 toxin groups during the pro-pregnancy of the middle-aged mice were [（219.56 ± 19.32）, （136.89 ± 20.41）, （210.49 ± 21.23）, （207.41 ± 21.23）; （192.73 ± 22.43）, （136.25 ± 29.55）, （187.43 ± 39.32）, （232.48 ± 39.32）; （1 303.02 ± 142.10）, （1 072.60 ± 78.30）, （1 065.03 ± 37.44）, and （1 169.72 ± 104.18） ng/L], respectively. The differences between control and T-2 toxin treated groups were statistically significant （F = 17.124, 6.237, 7.670, P 〈 0.05）. For further pairwise comparison, IL-1β and IL-6 in low dose group were significantly lower than those in control, middle and high dose groups （P 〈 0.05）; TNF-α content in control group was significantly higher than those in low, middle and high dose groups （P 〈 0.05）. There were significant differences in the levels of IL-1β, IL-6 and TNF-α between the control group and the low, middle and high dose groups of offspring weanling mice [（142.36 ± 13.36）, （113.01 ± 8.65）, （102.13 ± 8.31）, （123.42 ± 10.41）; （109.92 ± 9.76）, （100.26 ± 15.60）, （85.25 ± 9.97）, （100.21 ± 16.46）; （1 308.45 ± 204.90）, （1 248.60 ± 96.85）, （1 081.09 ± 105.51）, （1 204.87 ± 153.96） ng/L, F = 49.823, 10.530, 7.490, P 〈 0.05]. The levels of IL-1β and IL-6 in the control group were significantly higher than those in the low, middle and high dose groups （P 〈 0.05）. The levels of TNF-α in the control group were significantly higher than those in the medium and high dose groups （P 〈 0.05）. The levels of the three cytokines IL-1β, IL-6 and TNF-α in adult filial mice were significantly different [（69.71 ± 9.61）, （61.31 ± 10.07）, （63.07 ± 10.39）, （58.56 ± 9.69）; （172.55 ± 24.55）, （146.91 ± 13.47）, （151.02 ± 24.93）, （157.21 ± 17.86）; （1 136.87 ± 137.39）, （1 002.22 ± 86.52）, （987.12 ± 130.80）, （1 047.21 ± 171.64） ng/L, F = 4.670, 5.636, 4.775, P 〈 0.05], the contents of the three cytokines in the poisoning groups were significantly lower than that of the control group （P 〈 0.05）. The organ coefficients of thymus, spleen and liver in the second trimester were significantly different [（0.14 ± 0.03）, （0.20 ± 0.06）, （0.15 ± 0.02）, （0.12 ± 0.03）; （0.71 ± 0.16）, （0.78 ± 0.14）, （0.77 ± 0.15）, （0.38 ± 0.10）; （6.19 ± 0.43）, （5.57 ± 0.57）, （6.04 ± 0.32）, （5.11 ± 0.29）, F = 4.056, 11.064, 8.312, P 〈 0.05], and the thymus index was significantly increased in low dose group （P 〈 0.05）, spleen coefficient decreased significantly in high dose group （P 〈 0.05）, and liver coefficients in low and high dose group were significantly decreased （P 〈 0.05）. In the offspring, the midbrain coefficient of viscera showed significant changes [（3.45 ± 0.73）, （3.11 ± 0.31）, （2.98 ± 0.45）, （3.04 ± 0.22）, F = 7.529, P 〈 0.05], which was significantly decreased in the exposed rats （P 〈 0.05）. Both the mid-pregnant mice and filial mice showed varying degrees of changes in epiphyseal cartilage injury. The degree of epiphyseal cartilage injury became higher with increasing dosages of T-2 toxin in mid-pregnancy and post-weaning parental mice, and the injury was more serious in post-weaning mice.ConclusionsExposure to T-2 toxin can cause decrease of cytokines IL-1β, IL-6 and TNF-α in the blood of CD-1 pregnant and filial mice, and also cause the cartilage damage in mice, which are aggravated following increased doses of T-2 toxin and extension of exposure time.