斑马鱼功效模型在验证乳制品补钙和促进肠蠕动中的应用

Dairy products supplement calcium and promote intestinal motility in zebrafish models

目的基于斑马鱼补钙和肠蠕动模型,验证和对比不同牛奶补钙和酸奶促进肠蠕动的作用效果。方法选择发育健康且阶段一致的3 dpf野生型斑马鱼,加入不同浓度(2.5、5.0、10.0 mg/m L)的纯牛奶稀释液,同时设置空白对照组。处理至7 dpf后,用2 mg/m L钙黄绿素染色,在荧光显微镜下观察拍照,计算斑马鱼的椎骨荧光强度。选取5 dpf的斑马鱼,用尼罗红染料处理16 h,加入不同浓度(2.5、5.0、10.0 mg/m L)的酸奶和乳酸菌饮料稀释液,同时设置空白对照组。处理24 h后,在荧光显微镜下观察拍照,计算斑马鱼肠道内荧光强度。结果纯牛奶、酸奶和乳酸菌饮料作用效果都随浓度的增加而增强。与空白对照组比较,6种纯牛奶中2种纯牛奶处理的斑马鱼骨骼荧光强度有一定增加;8种酸奶和乳酸菌饮料都显著引起了肠道荧光强度的减弱,酸奶的效果优于乳酸菌饮料。结论纯牛奶、酸奶和乳酸菌饮料对斑马鱼的最大耐受浓度(MTC)为10 mg/m L,且对斑马鱼起到补钙和促进肠蠕动的作用。

Objective To validate and compare the functions of calcium supplementation and intestinal peristalsis promotion between two brands of dairy products using zebrafish models.Methods Zebrafish embryos at 3 days post fertilization(dpf)were selected and exposed to UHT(ultra heat treated)milk at 0(blank control),2.5,5.0,and 10.0 mg/m L for 96 h.After treatment,zebrafish were stained with calcein and observed under a stereofluorescent microscope to measure the fluorescence intensity of vertebrae.Zebrafish were also fed with Nile red at 5 dpf for 16 h,followed by a 24 h exposure to the same concentrations of yogurt and lactobacillus beverage.Intestinal tract fluorescence was measured to assess the ability of intestinal peristalsis promotion.Results UHT milk,yogurt,and lactobacillus beverage all increased the zebrafish vertebrate fluorescent intensity and intestinal motility in a concentration-dependent manner.Compared with the blank control,2 of the 6 types of UHT milk increased vertebrate fluorescent intensity;and 8 types of yogurt and lactobacillus beverage significantly elevated intestinal motility,although yogurt was better than the lactobacillus beverage.Conclusions The maximum tolerable concentration(MTC)of UHT milk,yogurt,and lactobacillus beverage is 10.0 mg/m L in zebrafish.These dairy products have the same functions in zebrafish as in humans in term of calcium supplementation and intestinal peristalsis effect.