酒精摄入导致雌鼠及孕鼠子代血清泌乳素升高及其机制的探讨
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摘要
第一部分酒精对雌鼠血清泌乳素水平及垂体湿重的影响
目的探讨酒精对雌鼠血清泌乳素(PRL)水平、垂体湿重的影响,了解摄取酒精的浓度和年龄与血清PRL水平改变及垂体增生的相关性,了解饮酒与高泌乳素血症(HP)的关系。
方法1.40只SD同龄成年雌鼠随机分为4组,每组分别予10%,20%,30%酒精灌胃4周和8周各5只,对照组予等容积和时间蒸馏水灌胃；2.30只4周、6周、8周龄的SD雌鼠分别予以20%酒精灌胃4周,各自对照组均予等容积蒸馏水灌胃；3.各组大鼠处死后均用放射免疫分析法(RIA)检测血清PRL水平,称垂体湿重,分析摄取酒精的浓度和年龄与血清PRL水平、垂体湿重的相关性。
结果1.灌胃4周时,30%酒精组大鼠血清PRL水平和垂体湿重明显高于其它三组；灌胃8周时,10%和20%酒精组明显高于同期30%酒精组和对照组。组内比较,灌胃8周时,10%及20%酒精组大鼠PRL水平和垂体湿重均显著高于灌胃4周时,30%酒精组大鼠PRL水平和垂体湿重显著低于4周时,以上各差异均有统计学意义,P<0.05。2.同浓度酒精灌胃4周后,各年龄组与相应的对照组比较,4周龄大鼠血清PRL水平和垂体湿重显著升高；6周龄和8周龄大鼠血清PRL水平、垂体湿重差异均无统计学意义,P>0.05。
结论1.酒精摄入可引起雌鼠血清PRL水平发生改变,且有剂量依赖性和累积效应：高浓度酒精可在较短时间内使血清PRL水平升高,长期低浓度酒精摄取可引起PRL水平显著性增高。2.长期大剂量酒精摄入可导致雌性大鼠垂体产生PRL的功能减退。3.较之成年期,幼年期雌性大鼠摄入酒精更易导致血清PRL水平升高。
第二部分摄入酒精对孕鼠子代血清泌乳素及垂体湿重的影响
目的探讨大鼠妊娠期大量摄入酒精对子代血清泌乳素水平、垂体湿重的影响。
方法6只孕鼠随机分为2组,酒精组于怀孕第1天(P1)起给予20%酒精(2g/kg·d)灌胃至分娩前。对照组同期给予等量蒸馏水。子鼠喂养至4周后每组随机抽取一半处死,余下的继续喂养至8周处死。各组大鼠处死后均用放免法检测血清泌乳素水平,并称垂体湿重。
结果1.4周龄子鼠：酒精组的垂体湿重明显高于对照组(P<0.05),而两组间PRL水平差异无统计学意义(P>0.05)；2.8周龄子鼠：酒精组的PRL水平和垂体湿重均较对照组显著升高(P<0.05)。
结论大鼠妊娠期大量摄入酒精可使子代血清泌乳素水平升高、垂体湿重增加。酒精可能通过刺激垂体增生,使得血泌乳素水平升高。
第三部分酒精摄入导致雌鼠血清泌乳素升高机制的探讨
目的：探讨酒精导致雌鼠血清泌乳素升高的机制。
方法：40只大鼠随机分为2组,实验组给予20%酒精2m1灌胃,每日2次共8周,对照组给予等体积蒸馏水灌胃。检测血清泌乳素水平、垂体湿重及免疫组化观察垂体腺细胞增殖情况,库伦阵列电化学高效液相色谱法检测下丘脑匀浆去甲肾上腺素、多巴胺、5-羟色胺、r-氨基丁酸、谷氨酸水平并进行比较。
结果：1.实验组血清泌乳素水平、垂体湿重均显著高于对照组,P<0.05。2.实验组下丘脑匀浆多巴胺、5-羟色胺、r-氨基丁酸显著低于对照组,谷氨酸浓度显著高于对照组, P<0.05。去甲肾上腺素浓度与对照组比较无显著性差异,P>0.05。3.实验组垂体细胞Ki-67的表达显著高于对照组,P<0.05。
结论：酒精可以导致泌乳素升高,其机制可能为：1.垂体催乳细胞增殖；2.酒精刺激引起下丘脑泌乳素释放抑制因子(PIF)如多巴胺、5-羟色胺、r-氨基丁酸等分泌减少,而泌乳素释放因子(PRF)如谷氨酸分泌增多。
Part 1:Effects of alcohol on the serum prolactin level, pituitary wet weights in female rats
Objective To discover the effect of alcohol on the serum prolactin (PRL) level and pituitary wet weights in female rats, understand the correlation between the alcohol concentration and the rats age and serum prolactin (PRL) level change and pituitary hyperplasia, and explore the relation between alcohol consumption and hyperprolactinemia (HP).
Methods 1. Forty grown female SD rats of the same age were randomly divided into three alcoholic groups and one control group, ten in each group.10%,20% and 30% ethanol were administered to three alcoholic groups respectively for 4 weeks (in 5 rats) and 8 weeks (in 5 rats) via gastrogavage; the control group was pair-fed on distilled water.2. 20% ethanol was administrated to thirty female SD rats aged four, six and eight weeks respectively for 4 weeks via gastrogavage, and the rats in the control groups were pair-fed on distilled water.3. The serum prolactin levels were detected using radioimmunoassay, the pituitary wet weights were measured, the correlation between the alcohol concentration and the rats age and the serum prolactin level and the pituitary wet weights was analysed.
Results 1. The serum prolactin levels and the pituitary wet weights in the group of 30% ethanol were significantly higher than that in other three groups after ethanol intake via gastrogavage for 4 weeks; the serum prolactin levels and the pituitary wet weights in the 10% and 20% ethanol groups were obviously higher than that in 30% ethanol group and the control group after alcohol consumption via gastrogavage for 8 weeks. The comparison made in the same group showed that the serum prolactin levels and the pituitary wet weights in the 10% and 20% ethanol groups after 8 weeks of alcohol consumption were remarkably higher than that in the same groups after 4 weeks of alcohol consumption; the serum prolactin levels and the pituitary wet weights in the 30% after 8 weeks of alcohol consumption were significantly higher that than in the same group after 4 weeks of alcohol consumption. The aforesaid differences were statistically significant, P<0.05.2. After 4 weeks of alcohol consumption with the same concentration,4-week-old rats presented significantly higher serum prolactin levels and the pituitary wet weights compared with respective control groups; while, there were no statistical significances between the serum prolactin levels and the pituitary wet weights in 6-week-old and 8-week-old rats. P>0.05.
Conclusion 1. Alcohol consumption can cause change in the serum prolactin level of female rats which depends on ethanol concentration and doses:Higher alcohol concentrations could increase the serum prolactin level within a relatively short time. Long-term consumption of lower alcohol concentrations may greatly elevate the plasma prolactin level.2. Long-term heavy alcohol consumption may lead to the pituitary dysfunction of secreting the prolactin.3. Compared with adult rats, a increased serum prolactin level could be more easily caused by alcohol consumption in juvenile rats.
Part 2 The Effect of High Alcohol Intake During Pregnancy on the Serum Prolactin Level and Pituitary Wet Weights of the Rat Progeny
Objective:To discuss the effects of high alcohol intake during pregnancy on the serum prolactin level and the pituitary wet weights of rat progeny
Methods:Six grown female SD rats of the same age were randomly divided into one alcoholic group and one control group, three in each group.20% alcohol was administrated to the rats in the alcohol group via gastrogavage (2g/kg per day) from the first day of pregnancy (P1) to pre-delivery. The rats in the control group were given distilled water in the same period. The offspring rats were fed for 4 weeks after birth, then half of the rats in each group were sacrificed at random. The remaining rats were fed continually for another 4 weeks before being killed. The prolactin levels of the rats in each group were detected using radioimmunoassay, and their pituitary wet weights were measured.
Results:1. With 4-week-old offspring:the pituitary wet weights in the alcohol group were significantly higher than that in the control group (P<0.05), while there was no statistical significance in the differences between the PRL levels in the two groups (P> 0.05); 2. With 8-week-old offspring:the pituitary wet weights and the PRL levels of the rats in the alcohol group were significantly higher than those in the control group (P <0.05).
Conclusion:High alcohol intake during pregnancy in rats could reduce progeny serum prolactin levels and increase the pituitary wet weights. Alcohol may cause a increased blood prolactin level by stimulating the pituitary hyperplasia.
Part 3 Study on Mechanism of the Serum Prolactin Increase in Female Rats Caused by Alcohol Exposure
Objective:Study on mechanism of a increased serum prolactin in female rats by alcohol exposure.
Methods:Forty grown female SD rats of the same age were randomly divided into one alcoholic group and one control group, twenty in each group. The rats were implanted with intragastric cannulae. The alcoholic group was fed on 2ml of 20% ethanol,2 times daily for 8 weeks. The control group was pair-fed on distilled water. The serum prolactin levels were detected, the pituitary wet weights were measured, and the pituitary gland cell proliferation were detected using immunohis-tochemistry. Norepinephrine, dopamine,5-HT, r-aminobutyric acid, glutamate in hypothalamus homogenate were assayed using coulomb array electrochemistry high performance liquid chromatography(HPLC) and the comparisons were made.
Results:1. The serum prolactin levels, the pituitary wet weights in the experimental group were significantly higher than that in the control group P<0.05.2. The hypothalamic homogenate dopamine,5-HT, r-aminobutyric acid in the experimental group were significantly lower than those in the control group. While, the glutamate acid concentration in the experimental group was significantly higher than that in the control group, P<0.05. There was no significant difference between noradrenaline concentrations in the two groups, P>0.05.3. The Ki-67 expression in the experimental group was remarkably higher than that in the control group, P<0.05.
Conclusion:Alcohol increases the serum prolactin, and the mechanism is probably associated with:1. the pituitary prolactin cell proliferation 2. a decreased secretion of prolactin release inhibiting factor (PIF) such as dopamine,5-HT, r-aminobutyric acid by the hypothalamus after alcohol stimulation, and a increased secretion of prolactin releasing factor (PRF), such as glutamate by the hypothalamus after alcohol stimulation.
目的探讨酒精对雌鼠血清泌乳素(PRL)水平、垂体湿重的影响,了解摄取酒精的浓度和年龄与血清PRL水平改变及垂体增生的相关性,了解饮酒与高泌乳素血症(HP)的关系。
方法1.40只SD同龄成年雌鼠随机分为4组,每组分别予10%,20%,30%酒精灌胃4周和8周各5只,对照组予等容积和时间蒸馏水灌胃；2.30只4周、6周、8周龄的SD雌鼠分别予以20%酒精灌胃4周,各自对照组均予等容积蒸馏水灌胃；3.各组大鼠处死后均用放射免疫分析法(RIA)检测血清PRL水平,称垂体湿重,分析摄取酒精的浓度和年龄与血清PRL水平、垂体湿重的相关性。
结果1.灌胃4周时,30%酒精组大鼠血清PRL水平和垂体湿重明显高于其它三组；灌胃8周时,10%和20%酒精组明显高于同期30%酒精组和对照组。组内比较,灌胃8周时,10%及20%酒精组大鼠PRL水平和垂体湿重均显著高于灌胃4周时,30%酒精组大鼠PRL水平和垂体湿重显著低于4周时,以上各差异均有统计学意义,P<0.05。2.同浓度酒精灌胃4周后,各年龄组与相应的对照组比较,4周龄大鼠血清PRL水平和垂体湿重显著升高；6周龄和8周龄大鼠血清PRL水平、垂体湿重差异均无统计学意义,P>0.05。
结论1.酒精摄入可引起雌鼠血清PRL水平发生改变,且有剂量依赖性和累积效应：高浓度酒精可在较短时间内使血清PRL水平升高,长期低浓度酒精摄取可引起PRL水平显著性增高。2.长期大剂量酒精摄入可导致雌性大鼠垂体产生PRL的功能减退。3.较之成年期,幼年期雌性大鼠摄入酒精更易导致血清PRL水平升高。
第二部分摄入酒精对孕鼠子代血清泌乳素及垂体湿重的影响
目的探讨大鼠妊娠期大量摄入酒精对子代血清泌乳素水平、垂体湿重的影响。
方法6只孕鼠随机分为2组,酒精组于怀孕第1天(P1)起给予20%酒精(2g/kg·d)灌胃至分娩前。对照组同期给予等量蒸馏水。子鼠喂养至4周后每组随机抽取一半处死,余下的继续喂养至8周处死。各组大鼠处死后均用放免法检测血清泌乳素水平,并称垂体湿重。
结果1.4周龄子鼠：酒精组的垂体湿重明显高于对照组(P<0.05),而两组间PRL水平差异无统计学意义(P>0.05)；2.8周龄子鼠：酒精组的PRL水平和垂体湿重均较对照组显著升高(P<0.05)。
结论大鼠妊娠期大量摄入酒精可使子代血清泌乳素水平升高、垂体湿重增加。酒精可能通过刺激垂体增生,使得血泌乳素水平升高。
第三部分酒精摄入导致雌鼠血清泌乳素升高机制的探讨
目的：探讨酒精导致雌鼠血清泌乳素升高的机制。
方法：40只大鼠随机分为2组,实验组给予20%酒精2m1灌胃,每日2次共8周,对照组给予等体积蒸馏水灌胃。检测血清泌乳素水平、垂体湿重及免疫组化观察垂体腺细胞增殖情况,库伦阵列电化学高效液相色谱法检测下丘脑匀浆去甲肾上腺素、多巴胺、5-羟色胺、r-氨基丁酸、谷氨酸水平并进行比较。
结果：1.实验组血清泌乳素水平、垂体湿重均显著高于对照组,P<0.05。2.实验组下丘脑匀浆多巴胺、5-羟色胺、r-氨基丁酸显著低于对照组,谷氨酸浓度显著高于对照组, P<0.05。去甲肾上腺素浓度与对照组比较无显著性差异,P>0.05。3.实验组垂体细胞Ki-67的表达显著高于对照组,P<0.05。
结论：酒精可以导致泌乳素升高,其机制可能为：1.垂体催乳细胞增殖；2.酒精刺激引起下丘脑泌乳素释放抑制因子(PIF)如多巴胺、5-羟色胺、r-氨基丁酸等分泌减少,而泌乳素释放因子(PRF)如谷氨酸分泌增多。
Part 1:Effects of alcohol on the serum prolactin level, pituitary wet weights in female rats
Objective To discover the effect of alcohol on the serum prolactin (PRL) level and pituitary wet weights in female rats, understand the correlation between the alcohol concentration and the rats age and serum prolactin (PRL) level change and pituitary hyperplasia, and explore the relation between alcohol consumption and hyperprolactinemia (HP).
Methods 1. Forty grown female SD rats of the same age were randomly divided into three alcoholic groups and one control group, ten in each group.10%,20% and 30% ethanol were administered to three alcoholic groups respectively for 4 weeks (in 5 rats) and 8 weeks (in 5 rats) via gastrogavage; the control group was pair-fed on distilled water.2. 20% ethanol was administrated to thirty female SD rats aged four, six and eight weeks respectively for 4 weeks via gastrogavage, and the rats in the control groups were pair-fed on distilled water.3. The serum prolactin levels were detected using radioimmunoassay, the pituitary wet weights were measured, the correlation between the alcohol concentration and the rats age and the serum prolactin level and the pituitary wet weights was analysed.
Results 1. The serum prolactin levels and the pituitary wet weights in the group of 30% ethanol were significantly higher than that in other three groups after ethanol intake via gastrogavage for 4 weeks; the serum prolactin levels and the pituitary wet weights in the 10% and 20% ethanol groups were obviously higher than that in 30% ethanol group and the control group after alcohol consumption via gastrogavage for 8 weeks. The comparison made in the same group showed that the serum prolactin levels and the pituitary wet weights in the 10% and 20% ethanol groups after 8 weeks of alcohol consumption were remarkably higher than that in the same groups after 4 weeks of alcohol consumption; the serum prolactin levels and the pituitary wet weights in the 30% after 8 weeks of alcohol consumption were significantly higher that than in the same group after 4 weeks of alcohol consumption. The aforesaid differences were statistically significant, P<0.05.2. After 4 weeks of alcohol consumption with the same concentration,4-week-old rats presented significantly higher serum prolactin levels and the pituitary wet weights compared with respective control groups; while, there were no statistical significances between the serum prolactin levels and the pituitary wet weights in 6-week-old and 8-week-old rats. P>0.05.
Conclusion 1. Alcohol consumption can cause change in the serum prolactin level of female rats which depends on ethanol concentration and doses:Higher alcohol concentrations could increase the serum prolactin level within a relatively short time. Long-term consumption of lower alcohol concentrations may greatly elevate the plasma prolactin level.2. Long-term heavy alcohol consumption may lead to the pituitary dysfunction of secreting the prolactin.3. Compared with adult rats, a increased serum prolactin level could be more easily caused by alcohol consumption in juvenile rats.
Part 2 The Effect of High Alcohol Intake During Pregnancy on the Serum Prolactin Level and Pituitary Wet Weights of the Rat Progeny
Objective:To discuss the effects of high alcohol intake during pregnancy on the serum prolactin level and the pituitary wet weights of rat progeny
Methods:Six grown female SD rats of the same age were randomly divided into one alcoholic group and one control group, three in each group.20% alcohol was administrated to the rats in the alcohol group via gastrogavage (2g/kg per day) from the first day of pregnancy (P1) to pre-delivery. The rats in the control group were given distilled water in the same period. The offspring rats were fed for 4 weeks after birth, then half of the rats in each group were sacrificed at random. The remaining rats were fed continually for another 4 weeks before being killed. The prolactin levels of the rats in each group were detected using radioimmunoassay, and their pituitary wet weights were measured.
Results:1. With 4-week-old offspring:the pituitary wet weights in the alcohol group were significantly higher than that in the control group (P<0.05), while there was no statistical significance in the differences between the PRL levels in the two groups (P> 0.05); 2. With 8-week-old offspring:the pituitary wet weights and the PRL levels of the rats in the alcohol group were significantly higher than those in the control group (P <0.05).
Conclusion:High alcohol intake during pregnancy in rats could reduce progeny serum prolactin levels and increase the pituitary wet weights. Alcohol may cause a increased blood prolactin level by stimulating the pituitary hyperplasia.
Part 3 Study on Mechanism of the Serum Prolactin Increase in Female Rats Caused by Alcohol Exposure
Objective:Study on mechanism of a increased serum prolactin in female rats by alcohol exposure.
Methods:Forty grown female SD rats of the same age were randomly divided into one alcoholic group and one control group, twenty in each group. The rats were implanted with intragastric cannulae. The alcoholic group was fed on 2ml of 20% ethanol,2 times daily for 8 weeks. The control group was pair-fed on distilled water. The serum prolactin levels were detected, the pituitary wet weights were measured, and the pituitary gland cell proliferation were detected using immunohis-tochemistry. Norepinephrine, dopamine,5-HT, r-aminobutyric acid, glutamate in hypothalamus homogenate were assayed using coulomb array electrochemistry high performance liquid chromatography(HPLC) and the comparisons were made.
Results:1. The serum prolactin levels, the pituitary wet weights in the experimental group were significantly higher than that in the control group P<0.05.2. The hypothalamic homogenate dopamine,5-HT, r-aminobutyric acid in the experimental group were significantly lower than those in the control group. While, the glutamate acid concentration in the experimental group was significantly higher than that in the control group, P<0.05. There was no significant difference between noradrenaline concentrations in the two groups, P>0.05.3. The Ki-67 expression in the experimental group was remarkably higher than that in the control group, P<0.05.
Conclusion:Alcohol increases the serum prolactin, and the mechanism is probably associated with:1. the pituitary prolactin cell proliferation 2. a decreased secretion of prolactin release inhibiting factor (PIF) such as dopamine,5-HT, r-aminobutyric acid by the hypothalamus after alcohol stimulation, and a increased secretion of prolactin releasing factor (PRF), such as glutamate by the hypothalamus after alcohol stimulation.
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