三氯生对孕鼠妊娠结局的影响及初步机理探讨

发布时间：2018-04-12 14:03

本文选题：三氯生 + 器官形成期　；参考：《兰州大学》2013年硕士论文

【摘要】：目的通过综合分析外源性三氯生对孕鼠妊娠结局的影响,进一步研究其对孕鼠食物利用率、激素水平、血糖水平、抗氧化能力及胎鼠脑组织凋亡蛋白表达等方面的影响,探讨影响妊娠结局的可能机制,为指导一般人群及孕妇合理使用含三氯生类产品(消杀、洗涤、日用品等)提供更确定的实验参考。方法将32只妊娠Wistar大鼠随机分为4组,溶剂对照(花生油)组和30mg/kg-d、100mg/kg-d、300mg/kg-d剂量组,妊娠6-20d连续灌胃给予相应受试物,孕20d处死,记录孕鼠的食物利用率、激素水平、血糖水平、抗氧化能力及胎鼠脑组织凋亡蛋白表达等指标。结果一、三氯生对孕鼠总体妊娠结局的影响 300mg/kg-d组孕鼠活胎和死胎的构成比例均低于对照组,吸收胎的构成比例高于对照组,但均无统计学差异(P0.05)；各组活胎鼠发育基本良好,无畸形表现,雌雄比差异无显著性(P0.05)；100mg/kg.d组及对照组胎鼠的囟门明显大于300mg/kg.d组(P0.05)。二、三氯生影响妊娠结局的可能机制 2.1三氯生对母体相关指标的影响情况 2.1.1从对不同孕期孕鼠进食量、体重、食物利用率的影响方面分析：实验期间各组孕鼠进食量、体重和食物利用率总体随孕期延长呈上升现象(P0.01),组间比较显示：300mg/kg.d组孕鼠孕中期进食量和食物利用率显著低于对照组(P0.05),不同孕期孕鼠体重在各组之间无统计学差异(P0.05),重复测量方差分析结果显示,不同孕期孕鼠进食量、体重和食物利用率在各组之间均差异明显(P0.01),且体重的时间因素和处理因素间存在交互作用(P0.01)。 2.1.2从对孕鼠的脏器系数的影响方面分析：300mg/kg.d组孕鼠胸腺系数明显低于对照组(P0.05),且随三氯生剂量的增加而降低(r=-0.40,P=0.019),其余各脏器系数在各组之间差异均无显著性(P0.05)。 2.1.3从对孕鼠肝肾功能及血脂的影响方面分析：300mg/kg.d组总蛋白水平明显低于对照组(P0.05),其余指标在各组间均无统计学差异。 2.1.4从对不同孕期孕鼠血糖水平的影响方面分析：各组孕鼠空腹血糖于孕12d略升高,孕19d略下降。组内比较显示,300mg/kg.d组孕鼠不同时间段空腹血糖间差异有显著性(P0.01)。组间比较不同时间点各组孕鼠空腹血糖均无显著差异(P0.05),重复测量方差分析结果显示,不同孕期孕鼠血糖在各组之间有统计学差异(P0.05),但时间因素与处理因素之间无交互作用。 2.1.5从对孕鼠血清抗氧化水平和甲状腺激素的影响方面分析：对照组孕鼠血清MDA水平显著高于100mg/kg.d和300mg/kg.d组(P0.01),并随三氯生剂量的增加逐渐降低(r=-0.659,P=0.003)；300mg/kg.d组孕鼠TT3水平明显低于对照组(P0.05),且随三氯生剂量的增加而降低(r=-0.708,P=0.015)。 2.2三氯生对胎盘相关指标的影响情况 2.2.1从对孕鼠胎盘重量的影响方面分析：100mg/kg.d组和300mg/kg.d组的胎盘重量明显低于对照组(P0.05),并且与三氯生剂量呈负相关(r=-0.369,P=0.019)。 2.2.2从对孕鼠胎盘抗氧化水平及TNF-a的影响方面分析：各组孕鼠胎盘抗氧化水平及TNF-a均无统计学差异(P0.05)。 2.2.3从对孕鼠胎盘Bax、 Bcl-2、 Caspase-3蛋白表达的影响方面分析：孕鼠胎盘滋养细胞、脱膜组织及内膜腺体的凋亡蛋白的表达在各组之间均无统计学意义(P0.05),细胞浆的染色较浅。 2.3三氯生对胚胎相关指标的影响情况 2.3.1从对胎鼠脑组织水分及抗氧化水平的影响方面分析：各组胎鼠脑组织水分间差异无显著性(P0.05)。300mg/kg.d组胎鼠脑组织总SOD. Cu-Zn SOD. GSH-Px水平明显低于对照组(P0.05),并且与三氯生的剂量呈负相关(r=-0.538,P=0.003；r=-0.513,P=0.005；r=-0.432,P=0.012),MDA水平与三氯生剂量无相关性(r=-0.056,P=0.764)。 2.3.2从对胎鼠脑组织Bax、 Bcl-2、 Caspase-3蛋白表达的影响方面分析：胎鼠脑组织中Bax、 Bcl-2、 Caspase-3蛋白表达主要集中在胞浆中,脑组织颗粒细胞中Bax蛋白表达在100mg/kg.d组明显高于对照组和300mg/kg.d组(P0.05)；胶质细胞中Bax与Bcl-2蛋白表达在300mg/kg.d组和100mg/kg.d组均高于对照组(P0.05)；脑组织中Caspase-3蛋白表达在各组之间差异均无显著性(P0.05)；胶质细胞中Bax、 Bcl-2与Caspase-3蛋白表达与三氯生的剂量呈正相关(r=0.628,P=0.029；r=0.825,P=0.001；r=0.660,P=0.020)。结论孕鼠妊娠期间大量接触含三氯生类产品对母体和胚胎都有一定的影响,且其危害随摄入剂量的增大越严重,主要表现在对孕鼠的食物利用率、胸腺系数、血糖、甲状腺激素水平、胎鼠脑组织抗氧化水平及凋亡蛋白表达等方面。提示人群日常生活中应该避免长期大剂量接触含三氯生类产品,尤其是孕妇,在器官形成期和围生期要合理选择消杀、洗涤、日常护理产品等。
[Abstract]:Objective to influence on pregnancy outcome in rats by comprehensive analysis of Exogenous Chlorine three, further study on the utilization of food in pregnant rats with hormone levels, blood sugar levels, effects of antioxidant ability and fetal brain tissue apoptosis protein expression, and explore the possible mechanism of affecting the outcome of pregnancy, pregnant women and the general population for guiding the rational use of containing three triclosan products (disinfection, washing, etc.) to provide experimental reference more determined.
Methods 32 pregnant Wistar rats were randomly divided into 4 groups (peanut oil), solvent control group and 30mg/kg-d, 100mg/kg-d, 300mg/kg-d groups, 6-20d orally given pregnancy test, blood glucose levels were recorded at 20d, the food utilization rate of pregnant rats, hormone levels, and antioxidant capacity of fetal rats the brain tissue apoptosis protein expression and other indicators.
Result
The effect of three chloro on the overall pregnancy outcome of pregnant rats
The proportion of pregnant rats of 300mg/kg-d group live births and stillbirths were lower than the control group, the proportion of fetal absorption was higher than the control group, but there was no significant difference (P0.05); group of live fetal development is good, no deformity, no significant difference between the male and female ratio (P0.05); 100mg/kg.d group and control group in fetal rat fontanelle the group was significantly higher than that of 300mg/kg.d (P0.05).
The possible mechanism of two, three chloro - induced pregnancy outcome
2.1 effect of three chloro on parent related indexes
From the 2.1.1 of different pregnancy pregnant rats food intake, body weight, food utilization effect analysis: during the experiment groups of pregnant rats food intake, body weight and food utilization rate increased with prolonged pregnant rising phenomenon (P0.01), group comparison: 300mg/kg.d group of pregnant rats in mid pregnancy food consumption and food utilization rate was significantly lower than that of control group (P0.05), there was no significant difference in body weight between the groups in different pregnant rats (P0.05), repeated measurement of variance analysis showed that different pregnancy pregnant rats food intake, body weight and food utilization rate among the groups were significantly different (P0.01), and there was an interaction between the weight of the time factor and processing factors (P0.01).
2.1.2 analyzed the influence of the organ coefficient on the pregnant rats: the thymus coefficient in the 300mg/kg.d group was significantly lower than that in the control group (P0.05), and decreased with the increase of three chlorate birth dose (r=-0.40, P=0.019), and there was no significant difference in the other organ coefficients between the groups (P0.05).
The effect of 2.1.3 on the function of liver and kidney and blood lipid in pregnant rats was analyzed. The total protein level in group 300mg/kg.d was significantly lower than that in control group (P0.05), and there was no significant difference in other indicators among groups.
2.1.4 from the influence on the blood glucose level in rats during pregnancy: analysis of different groups of fasting blood glucose in pregnant rats on gestation 12D increased slightly at 19d decreased slightly. Within group comparison, there was significant in different time group 300mg/kg.d pregnant rats fasting blood glucose (P0.01). The differences between different groups were no significant differences in fasting time the mice blood glucose (P0.05), repeated measurement of variance analysis showed that there were significant differences between the groups in different pregnant rats blood glucose (P0.05), but no interaction between time factors and processing factors.
2.1.5 from the aspect of pregnant rats serum antioxidant levels and thyroid hormone analysis: control group rats serum MDA level was significantly higher than that of 100mg/kg.d and 300mg/kg.d group (P0.01), and decreased with the increase of three triclosan dose decreased (r=-0.659, P=0.003); the level of TT3 in pregnant rats of 300mg/kg.d group was significantly lower than the control group (P0.05), and with the increase of three triclosan dose decreased (r=-0.708, P=0.015).
2.2 effect of three chloro on placental related indexes
2.2.1 analyzed the influence of placental weight on pregnant rats: placental weight in group 100mg/kg.d and 300mg/kg.d was significantly lower than that in control group (P0.05), and negatively correlated with the dose of three chloro chloride (r=-0.369, P=0.019).
The effect of 2.2.2 on the antioxidation level of placenta and the effect of TNF-a on the placenta of pregnant rats were analyzed: there was no significant difference in the antioxidant level and TNF-a of placenta of pregnant rats (P0.05).
The effect of 2.2.3 on the expression of Bax, Bcl-2 and Caspase-3 protein in placenta of pregnant rats was analyzed. The expression of apoptotic protein in placental trophoblast, endometrial glands and endometrial glands in pregnant rats was not statistically significant among all groups (P0.05), and the staining of cytoplasm was shallow.
2.3 effect of three chloro on embryo related indexes
2.3.1 from the aspect of water and antioxidant levels in fetal rat brain: analysis of differences between water content in brain tissue of fetal rats were not significant (P0.05) group.300mg/kg.d fetal rat brain total SOD. Cu-Zn SOD. GSH-Px was significantly lower than the control group (P0.05), and negatively correlated with the dose of three triclosan (r=-0.538, P=0.003 P=0.005; r=-0.432; r=-0.513, P=0.012, MDA) level and three triclosan dose no correlation (r=-0.056, P=0.764).
2.3.2 from Bcl-2 on fetal rat brain Bax, analysis of Caspase-3 protein expression in the brain tissue of fetal rats in Bax, Bcl-2, Caspase-3 protein was mainly expressed in the cytoplasm, were significantly higher in the 100mg/kg.d group and 300mg/ kg.d group the expression of Bax protein in brain tissue in granulosa cells (P0.05) in the 300mg/kg.d group; and the 100mg/kg.d group were higher than the control group the expression of Bax and Bcl-2 protein in glial cells (P0.05); the expression of Caspase-3 protein in brain tissue in each group. There was no significant difference (P0.05); glial cells in Bax, Bcl-2 and Caspase-3 protein expression was positively correlated with the dosage of three triclosan (r=0.628, P=0.029; r=0.825, P=0.001; r=0.660, P=0.020).
Conclusion during pregnancy in pregnant rats exposed to triclosan containing three categories of products have certain effects on maternal and embryo, and its harm with the increase of intake is more serious, mainly in the pregnant rat food utilization rate, thymus index, blood glucose, thyroid hormones, fetal rat brain tissue antioxidant levels and expression of apoptosis protein etc. suggests that people in daily life should avoid long-term large doses containing three triclosan products, especially in pregnant women, the formation period and the perinatal period should be a reasonable choice of disinfection, washing in organs, daily care products.

【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R114

【参考文献】