發(fā)布時(shí)間：2018-04-12 14:03

  本文選題：三氯生 + 器官形成期��； 參考：《蘭州大學(xué)》2013年碩士論文

【摘要】：目的通過綜合分析外源性三氯生對(duì)孕鼠妊娠結(jié)局的影響,進(jìn)一步研究其對(duì)孕鼠食物利用率、激素水平、血糖水平、抗氧化能力及胎鼠腦組織凋亡蛋白表達(dá)等方面的影響,探討影響妊娠結(jié)局的可能機(jī)制,為指導(dǎo)一般人群及孕婦合理使用含三氯生類產(chǎn)品(消殺、洗滌、日用品等)提供更確定的實(shí)驗(yàn)參考。 方法將32只妊娠Wistar大鼠隨機(jī)分為4組,溶劑對(duì)照(花生油)組和30mg/kg-d、100mg/kg-d、300mg/kg-d劑量組,妊娠6-20d連續(xù)灌胃給予相應(yīng)受試物,孕20d處死,記錄孕鼠的食物利用率、激素水平、血糖水平、抗氧化能力及胎鼠腦組織凋亡蛋白表達(dá)等指標(biāo)。 結(jié)果 一、三氯生對(duì)孕鼠總體妊娠結(jié)局的影響 300mg/kg-d組孕鼠活胎和死胎的構(gòu)成比例均低于對(duì)照組,吸收胎的構(gòu)成比例高于對(duì)照組,但均無統(tǒng)計(jì)學(xué)差異(P0.05)；各組活胎鼠發(fā)育基本良好,無畸形表現(xiàn),雌雄比差異無顯著性(P0.05)；100mg/kg.d組及對(duì)照組胎鼠的囟門明顯大于300mg/kg.d組(P0.05)。 二、三氯生影響妊娠結(jié)局的可能機(jī)制 2.1三氯生對(duì)母體相關(guān)指標(biāo)的影響情況 2.1.1從對(duì)不同孕期孕鼠進(jìn)食量、體重、食物利用率的影響方面分析：實(shí)驗(yàn)期間各組孕鼠進(jìn)食量、體重和食物利用率總體隨孕期延長呈上升現(xiàn)象(P0.01),組間比較顯示：300mg/kg.d組孕鼠孕中期進(jìn)食量和食物利用率顯著低于對(duì)照組(P0.05),不同孕期孕鼠體重在各組之間無統(tǒng)計(jì)學(xué)差異(P0.05),重復(fù)測量方差分析結(jié)果顯示,不同孕期孕鼠進(jìn)食量、體重和食物利用率在各組之間均差異明顯(P0.01),且體重的時(shí)間因素和處理因素間存在交互作用(P0.01)。 2.1.2從對(duì)孕鼠的臟器系數(shù)的影響方面分析：300mg/kg.d組孕鼠胸腺系數(shù)明顯低于對(duì)照組(P0.05),且隨三氯生劑量的增加而降低(r=-0.40,P=0.019),其余各臟器系數(shù)在各組之間差異均無顯著性(P0.05)。 2.1.3從對(duì)孕鼠肝腎功能及血脂的影響方面分析：300mg/kg.d組總蛋白水平明顯低于對(duì)照組(P0.05),其余指標(biāo)在各組間均無統(tǒng)計(jì)學(xué)差異。 2.1.4從對(duì)不同孕期孕鼠血糖水平的影響方面分析：各組孕鼠空腹血糖于孕12d略升高,孕19d略下降。組內(nèi)比較顯示,300mg/kg.d組孕鼠不同時(shí)間段空腹血糖間差異有顯著性(P0.01)。組間比較不同時(shí)間點(diǎn)各組孕鼠空腹血糖均無顯著差異(P0.05),重復(fù)測量方差分析結(jié)果顯示,不同孕期孕鼠血糖在各組之間有統(tǒng)計(jì)學(xué)差異(P0.05),但時(shí)間因素與處理因素之間無交互作用。 2.1.5從對(duì)孕鼠血清抗氧化水平和甲狀腺激素的影響方面分析：對(duì)照組孕鼠血清MDA水平顯著高于100mg/kg.d和300mg/kg.d組(P0.01),并隨三氯生劑量的增加逐漸降低(r=-0.659,P=0.003)；300mg/kg.d組孕鼠TT3水平明顯低于對(duì)照組(P0.05),且隨三氯生劑量的增加而降低(r=-0.708,P=0.015)。 2.2三氯生對(duì)胎盤相關(guān)指標(biāo)的影響情況 2.2.1從對(duì)孕鼠胎盤重量的影響方面分析：100mg/kg.d組和300mg/kg.d組的胎盤重量明顯低于對(duì)照組(P0.05),并且與三氯生劑量呈負(fù)相關(guān)(r=-0.369,P=0.019)。 2.2.2從對(duì)孕鼠胎盤抗氧化水平及TNF-a的影響方面分析：各組孕鼠胎盤抗氧化水平及TNF-a均無統(tǒng)計(jì)學(xué)差異(P0.05)。 2.2.3從對(duì)孕鼠胎盤Bax、 Bcl-2、 Caspase-3蛋白表達(dá)的影響方面分析：孕鼠胎盤滋養(yǎng)細(xì)胞、脫膜組織及內(nèi)膜腺體的凋亡蛋白的表達(dá)在各組之間均無統(tǒng)計(jì)學(xué)意義(P0.05),細(xì)胞漿的染色較淺。 2.3三氯生對(duì)胚胎相關(guān)指標(biāo)的影響情況 2.3.1從對(duì)胎鼠腦組織水分及抗氧化水平的影響方面分析：各組胎鼠腦組織水分間差異無顯著性(P0.05)。300mg/kg.d組胎鼠腦組織總SOD. Cu-Zn SOD. GSH-Px水平明顯低于對(duì)照組(P0.05),并且與三氯生的劑量呈負(fù)相關(guān)(r=-0.538,P=0.003；r=-0.513,P=0.005；r=-0.432,P=0.012),MDA水平與三氯生劑量無相關(guān)性(r=-0.056,P=0.764)。 2.3.2從對(duì)胎鼠腦組織Bax、 Bcl-2、 Caspase-3蛋白表達(dá)的影響方面分析：胎鼠腦組織中Bax、 Bcl-2、 Caspase-3蛋白表達(dá)主要集中在胞漿中,腦組織顆粒細(xì)胞中Bax蛋白表達(dá)在100mg/kg.d組明顯高于對(duì)照組和300mg/kg.d組(P0.05)；膠質(zhì)細(xì)胞中Bax與Bcl-2蛋白表達(dá)在300mg/kg.d組和100mg/kg.d組均高于對(duì)照組(P0.05)；腦組織中Caspase-3蛋白表達(dá)在各組之間差異均無顯著性(P0.05)；膠質(zhì)細(xì)胞中Bax、 Bcl-2與Caspase-3蛋白表達(dá)與三氯生的劑量呈正相關(guān)(r=0.628,P=0.029；r=0.825,P=0.001；r=0.660,P=0.020)。 結(jié)論孕鼠妊娠期間大量接觸含三氯生類產(chǎn)品對(duì)母體和胚胎都有一定的影響,且其危害隨攝入劑量的增大越嚴(yán)重,主要表現(xiàn)在對(duì)孕鼠的食物利用率、胸腺系數(shù)、血糖、甲狀腺激素水平、胎鼠腦組織抗氧化水平及凋亡蛋白表達(dá)等方面。提示人群日常生活中應(yīng)該避免長期大劑量接觸含三氯生類產(chǎn)品,尤其是孕婦,在器官形成期和圍生期要合理選擇消殺、洗滌、日常護(hù)理產(chǎn)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.

【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R114

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