Effect of dietary supplementation of fish oil on intestinal barrier function in mouse fed with high-fat diet
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摘要:目的
研究日粮添加鱼油对高脂日粮饲喂小鼠肠道屏障功能的影响。
方法选用36只4周龄C57BL/6J雌性小鼠，随机分为对照组、高脂组、高脂+鱼油组，每组12只小鼠，分别饲喂基础日粮、高脂日粮、高脂日粮添加质量分数为5%的鱼油(等能替换高脂日粮中脂肪)。试验持续21周，每周测定小鼠采食量和体质量，期间测定小鼠肠道通透性以及粪便粗脂肪含量和能值；试验结束后，检测小鼠血清内毒素水平，检测小鼠肠道形态、杯状细胞的数量、肠道紧密连接蛋白和炎症因子的表达。
结果与高脂组相比，日粮添加鱼油显著提高了小鼠采食量和能量摄入、降低了粪便中粗脂肪的含量和能量排出，显著降低了小鼠体质量(P<0.05)。在肠道形态方面，与高脂组相比，添加鱼油使小鼠空肠和回肠的绒毛高度与隐窝深度比值(lV/dC)分别提高了43.1%和67.5%，使回肠绒毛杯状细胞的数量增多了16.7%(P<0.05)。与高脂组相比，添加鱼油使血清荧光葡聚糖和内毒素水平分别降低了34.3%和50.4%(P<0.05)，并逆转了高脂日粮造成的肠道紧密连接蛋白表达的降低。在炎症因子表达方面，与高脂组相比，添加鱼油显著降低了空肠和回肠内促炎因子IL-8、IL-6和IL-1β的表达，同时显著增加了抗炎因子IL-10的表达(P<0.05)。
结论日粮添加鱼油可降低由高脂日粮导致的小鼠肠道屏障功能损伤，这可能与鱼油降低肠道炎症有关。
Abstract:ObjectiveTo investigate the effect of dietary supplementation of fish oil on the barrier function of mouse fed with high-fat diet (HFD).
Method36 C57BL/6J female mice (4-week-old) were randomly divided into control group (CK), HFD group, and HFD +fish oil group, with 12 mice in each group. Mice in three groups were fed with basal diet, HFD and HFD supplemented with 5% fish oil (iso caloric with HFD), respectively. The experiment lasted for 21 weeks, and the food intake and body weight of mice were measured weekly. During the experiment, intestinal permeability of each mouse was measured, and the fat content and energy in feces were determined. At the end of the experiment, endotoxin levels in serum were detected, and intestinal morphology, the number of intestinal goblet cells, and the expressions of intestinal tight junction proteins and inflammatory factors were examined.
ResultCompared with HFD group, addition of fish oil in diet significantly increased food intake and energy intake, decreased fecal crude fat content and energy excretion, while significantly reduced body weight of mice (P<0.05). In terms of intestinal morphology, compared with HFD group, addition of fish oil significantly increased the ratio of villus height to crypt depth (lV/dC) by 43.1% in mouse jejunum and 67.5% in ileum and the number of goblet cells in ileum villous by 16.7% (P<0.05). Compared with HFD group, addition of fish oil reduced the fluorescent glucan and endotoxin levels in serum by 34.3% and 50.4% respectively (P<0.05). Addition of fish oil also reversed the decrease in expressions of intestinal tight junction proteins caused by HFD. In terms of expression of inflammation factors, compared with HFD group, addition of fish oil significantly decreased the expressions of proinflammatory factors includingIL-8, IL-6 and IL-1β, and significantly elevated the expression of anti-inflammatory factor IL-10 in mouse jejunum and ileum (P<0.05).
ConclusionDietary supplementation of fish oil can reduce the HFD-induced impairment of intestinal barrier function, which might be associated with the fish oil reducing intestinal inflammation.
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Keywords:
- mouse /
- fish oil /
- high-fat diet /
- intestinal barrier /
- tight junction protein /
- inflammation
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图 1 日粮添加鱼油对小鼠生长性状的影响
各图中，柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 1. Effect of dietary supplementation of fish oil on mouse growth
In each graph, different lowercase letters on bars indicate significant differences(P<0.05，Duncan’s method)
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图 2 日粮添加鱼油对小鼠空肠和回肠绒毛形态的影响
Figure 2. Effect of dietary supplementation of fish oil on the villus morphology of mouse jejunum and ileum
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图 3 日粮添加鱼油对小鼠空肠和回肠的绒毛高度与隐窝深比值(lV/dC)的影响
各图中，柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 3. Effect of dietary supplementation of fish oil on the ratio of villus length to crypt depth (lV/dC) in mouse jejunum and ileum
In each graph, different lowercase letters on bars indicate significant differences(P<0.05, Duncan’s method)
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图 5 日粮添加鱼油对小鼠回肠杯状细胞数量的影响
柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 5. Effect of dietary supplementation of fish oil on goblet cell number in mouse ileum
Different lowercase letters on bars indicate significant differences (P<0.05, Duncan’s method)
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图 6 日粮添加鱼油对小鼠血清中荧光葡聚糖和内毒素含量的影响
各图中，柱子上方的不同小写字母表示差异显著(P<0.05，Duncan’s法)
Figure 6. Effects of dietary supplementation of fish oil on fluorescent glucan and endotoxin contents in mouse serum
In each graph, different lowercase letters on bars indicate significant differences(P<0.05, Duncan’s method)
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图 7 小鼠空肠和回肠紧密连接蛋白的Western blot检测结果
Figure 7. Western blot results of tight junction proteins in mouse jejunum and ileum
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图 8 日粮添加鱼油对小鼠空肠和回肠紧密连接蛋白表达的影响
各图中，相同蛋白柱子上方的不同小写字母表示组间差异显著(P<0.05，Duncan’s法)
Figure 8. Effect of dietary supplementation of fish oil on the expression of tight junction protein in mouse jejunum and ileum
In each graph, different lowercase letters on bars of the same protein indicate significant differences among groups(P<0.05, Duncan’s method)
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图 9 日粮添加鱼油对小鼠空肠和回肠炎症因子表达的影响
各图中，相同基因柱子上方的不同小写字母表示组间差异显著(P<0.05，Duncan’s法)
Figure 9. Effect of dietary supplementation of fish oil on the expression of inflammatory factor in mouse jejunum and ileum
In each graph, different lowercase letters on bars of the same gene indicate significant differences among groups(P<0.05, Duncan’s method)
表 1 小鼠日粮成分组成
Table 1 The dietary composition for mice
项目
Item组成
Component对照组
CK group高脂组
HFD group高脂+鱼油组
HFD+ fish oil group原料及质量分数/(g·kg−1)
Ingredient and content玉米淀粉 Corn starch 397.5 0 0 酪蛋白 Casein 200.0 261.7 261.7 麦芽糖糊精 Maltodextrin 132.0 163.6 163.6 蔗糖 Sucrose 100.0 89.8 89.8 豆油 Soybean oil 70.0 32.7 32.7 猪油 Lard 0 320.6 273.7 纤维素 Cellulose 50.0 65.4 65.4 混合矿物质 Mineral Mix 35.0 45.8 45.8 混合维生素 Vitamin Mix 10.0 13.1 13.1 L−胱氨酸 L−cystine 3.0 3.9 3.9 氯化胆碱 Choline chloride 2.5 3.3 3.3 鱼油 Fish Oil 0 0 50.0 合计 Total 1 000.0 999.9 1 003.0 营养成分及质量分数/%
Nutrient and content蛋白 Protein 17.9 26.2 26.2 脂肪 Fat 7.0 34.9 34.9 碳水化合物 Carbohydrate 64.4 26.3 26.3 总能/(kJ·g−1)
Total energy15.07 21.93 21.93 
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表 2 小鼠炎症因子引物序列
Table 2 Sequences for primers of mouse inflammation factors
基因
Gene引物序列(5′→3′)
Primer sequence产物长度/bp
Product lengthIL-1β F：TCTCTGGCCTTGGAACATAGTCT
R：CAAAGCCATCAGTGAACTCATCA145 IL-6 F：TACCACTCCCAACAGACCTG
R：CAAGTGCATCATCGTTGTTCA135 IL-8 F：CACCCTCTGTCACCTGCTCAA
R：ATGGCGCTGAGAAGACTTGGT101 IL-10 F：CGGGAAGACAATAACTGCACCC
R：CAAAGCCATCAGTGACCTAATCA130 TNF-α F：AGGCACTCCCCCAAAAGATG
R：CAAAGCCATCAGTGACCTAATCA145 β-actin F：TCTCTGTCCTTGGAACATAGTCT
R：CAAAGCCATCAGTGACCTAATCA142
下载: 导出CSV
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