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妊娠期和哺乳期酒精效应的研究设计注意事项
About Experimental Research in Effects of Alcohol during Gestation and Lactation

一、动物分组的要求


除了喂养酒精液体饲料组和对照液体饲料之外,还应当设计一个标准饲料组。因此,如果研究中不可能酒精的剂量效应关系,至少需要这三组同时进行妊娠的实验。

设计这个饲料喂养组的作用是:

由于妊娠或哺乳期间,酒精的作用包括对饲料摄入量(饮食摄入量)和酒精的作用两方面。为了消除饲料摄入量的改变所致的效应,就需要将对照液体饲料组采用配对喂养法。由于酒精组摄入量一般是减少的,对照饲料组必然也就要强制减少摄入量,显然,这个对照组营养状况发生了改变,那么,这方面的改变是否影响妊娠母鼠的机能、胎儿?是否影响哺乳期母鼠或仔鼠?这些问题的答案可以通过与标准饲料喂养组进行比较而得以回答。

换句话说,当3组进行比较时,酒精组的效应=酒精的效应+(标准组的效应-酒精对照组的效应)

二、动物准备


(1)选用处女鼠。小鼠一般不小于18g,大鼠一般可以选用70~85天龄。

(2)确保动物营养状态正常。为此,应当采用符合动物营养需求的饲料进行预先喂养,让动物在营养平衡的饲料喂养后进行妊娠。

三、饲料准备


(1)标准饲料:南通特洛菲饲料科技有限公司饲料代码LAD3001G.

(2)酒精液体饲料和对照饲料

可以从南通特洛菲饲料科技有限公司购买以下模型饲料之一,点击超链接可以了解其特点:

饲料代码:TP4035:Lieber-DeCarli酒精液体饲料25%高蛋白型

饲料代码:TP4040:Weinberg-Keiver酒精液体饲料

饲料代码:TP4100:Yeh-Cerklewski酒精液体饲料

饲料代码:TP4230:Sahchis酒精液体饲料

饲料代码:TP4200:Wiener酒精液体饲料

酒精液体饲料中酒精热量(浓度):

应当根据研究需要和动物承受能力综合决定。如果过高,动物难以承受,过低则可能效应不明显。一般来说,小鼠用28%左右是可以的,大鼠可以稍高。

四、动物妊娠方法和妊娠期的处理


(1)动物用标准饲料喂养至少一周,平衡营养。

(2)处女鼠出嫁(交配:mating):将处女鼠放入独居的雄鼠笼内过夜。

(3)检查精液栓:次日早晨检查精液栓。如何发现有精液栓,该天命名为GD1(妊娠第1天,gestation day 1),称重,并将该母鼠标记为所在组别(用作哪一组)。

(4)每日检查母鼠状态,包括称重,异常情况,死亡情况。

(5)如果不是研究酒精对受妊的影响,酒精液体饲料或对照液体饲料一般是在妊娠的GD7-GD8给予酒精液体饲料喂养,如果过早,将可能导致妊娠终止或死胎。

(6)每天检查母鼠的情况,包括称重,异常情况,死亡情况。对于死亡母鼠应当解剖观察胚胎情况。

(7)从GD19开始每天注意关注分娩(正常一般在GD21分娩),一旦分娩,该天命名为PND0(postnatal day 0,出生当天),给予称重,记录出生数(雌性和雄性各多少)、存活数数和死亡数。

五、哺乳期酒精喂养的安排


为了观察哺乳期幼鼠的酒精效应,应当从新生鼠中挑选正常的新生鼠,挑选时要注意:

(1)从每窝中挑选的雌性和雄性数量相同。

(2)每组数量尽可能相近,并且要有足够数量的样本量,由于涉及死亡率观察,样本量不能过少。

从PND0开始进入哺乳期,母鼠继续喂养酒精液体饲料或对照液体饲料。为了观察不同阶段的酒精效应,可以将哺乳期分为以下三个阶段进行观察:

PND1-8: 第1阶段;

PND9-16:第2阶段;

PND17-24:第3阶段。

在上面每一期每一天观察:

(1)幼鼠情况:体重改变情况,死亡情况,等等。

(2)母鼠情况:体重改变情况,食物摄入量,“食仔”情况,上午7~8时尾静脉取血分析血液酒精浓度,等等。

如果研究胎儿酒精暴露(Fetal Alcohol Exposure, FAE)包括一系列胎儿疾病,最为典型的是胚胎酒精综合征(Fetal Alcohol Syndrome,FAS)、胎儿酒精谱系障碍(Fetal Alcohol Spectrum Disorders,FASDs)、酒精性胚胎神经发育异常(Alcohol-Related Neurodevelopmental Disorder,ARND),请点击继续阅读:胎儿酒精暴露、胚胎酒精综合征、胚胎酒精效应、酒精性胚胎神经发育异常,等疾病的研究方法

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有困惑?那就商量呗!

液体饲料的优点,你可以充分运用!

---------《》-------

marker可以任意缺乏或过载某营养素

marker可以任意添加药物或测试成分

marker可以精确定量饲料摄入量

marker可以任意定制饲料


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