高脂日粮对初情期小鼠乳腺发育的影响及分子机制

孟莹莹; 张静; 张枫琳; 艾玮; 朱晓彤; 束刚; 王丽娜; 高萍; 江青艳; 王松波

doi:10.7671/j.issn.1001-411X.2017.03.002

高脂日粮对初情期小鼠乳腺发育的影响及分子机制

华南农业大学动物科学学院，广东广州 510642

基金项目:

广东省科技计划 2015B020203005

广东省科技计划 2015A020224039

广东省现代农业科技创新联盟建设项目 2016LM2150

广州市珠江科技新星专项 201610010073

详细信息

作者简介:
孟莹莹(1989—)，男，硕士研究生，E-mail:1326036268@qq.com

通讯作者:
王松波(1980—)，男，副教授，博士，E-mail:songbowang@scau.edu.cn

中图分类号: S828
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出版历程
- 收稿日期: 2016-07-16
- 网络出版日期: 2023-05-17
- 刊出日期: 2017-05-09

Effect of high-fat diet on mammary gland development of pubertal mice and the molecular mechanism

College of Animal Science, South China Agricultural University, Guangzhou 510642, China

摘要

摘要:
目的
研究高脂日粮对初情期小鼠乳腺发育的影响及可能机制。
方法
选用20只4周龄昆明雌性小鼠，设对照组和高脂日粮组，试验期4周，每周称体质量并统计采食量。试验结束后采集小鼠乳腺并称质量，采用whole-mount及HE染色观察小鼠乳腺组织形态结构及导管数量。利用Western blot检测初情期小鼠乳腺组织胰岛素样生长因子1(Insulin-like growth factor1, IGF-1)、增殖相关信号通路关键蛋白蛋白激酶B(Protein kinase B, Akt)、胞外调控的蛋白激酶(Extracellular regulated protein kinase, ErK)以及炎症信号通路关键蛋白Toll样受体4(Toll-like receptor 4, TLR4)、c-Jun氨基末端激酶(c-Jun N-terminal kinase, JNK)和核因子κB激酶抑制剂(Inhibitor of nuclear factor kappa-B kinase, IKK)的表达。
结果
高脂日粮组小鼠采食量极显著低于对照组(P < 0.01)，但在能量摄入方面与对照组无显著差异。高脂日粮组小鼠的体质量增加显著高于对照组(P < 0.05)。高脂日粮组小鼠在乳腺质量、乳腺质量与体质量的比值上均极显著高于对照组(P < 0.01)。乳腺组织的whole-mount及HE染色结果表明，高脂日粮组小鼠乳腺导管发育明显被抑制，其乳腺组织的导管数和密度明显降低，乳腺组织的终末乳芽数量也极显著低于对照组(P < 0.01)。高脂日粮能够显著抑制小鼠乳腺组织IGF-I的蛋白表达以及Akt、Erk和IKK的磷酸化水平(P < 0.01或0.05)，极显著促进小鼠乳腺组织TLR4的蛋白表达(P < 0.01) 和显著促进JNK的磷酸化水平(P < 0.05)。
结论
高脂日粮能够抑制初情期小鼠乳腺发育，这可能是通过抑制乳腺增殖的相关信号和促进乳腺组织炎症的相关信号来实现。
- 高脂日粮 /
- 小鼠 /
- 乳腺发育 /
- 增殖 /
- 炎症 /
- 蛋白表达
Abstract:
Objective
To investigate the effect of high-fat diet (HFD) on mammary gland development of pubertal mice and explore the possible underlying mechanisms.
Method
Twenty 4-week-old Kunming female mice were randomly divided into control group and HFD group. The trial lasted for four weeks. The body weight and feed intake were recorded weekly. At the end of the trial, mammary glands were collected and weighed. The morphology and number of ducts were observed by using whole-mount and HE staining. Western blot was used to analyze the expressions of insulin-like growth factor 1 (IGF-1), proliferation related signaling pathway proteins such as protein kinase B (Akt) and extracellular regulated protein kinases (ErK), as well as inflammation related signaling pathway proteins, including toll-like receptor 4 (TLR4), c-Jun N-terminal kinase (JNK) and inhibitor of nuclear factor kappa-B kinase (IKK) in the mammary glands.
Result
The feed intake of the HFD group was significantly lower than that of control group(P < 0.05), There was no significant difference in energy intakes between two groups. The body weight gain of HFD group was significantly higher than that of control group (P < 0.05). In addition, HFD significantly increased mammary gland weight and the ratio of mammary gland weight to body weight (P < 0.01). The results of whole-mount and HE staining demonstrated that the mammary gland development of HFD group mice were markedly inhibited, with significantly decreased duct number, duct density and number of terminal end buds (P < 0.01). HFD significantly inhibited IGF-1 expression and the phosphorylation of Akt, Erk and IKK in mammary glands (P < 0.01 or 0.05), while significantly promoted TLR4 expression (P < 0.01) and JNK phosphorylation (P < 0.05).
Conclusion
HFD can suppress mammary gland development of pubertal mice, which might result from the inhibiton of proliferative signals and stimulation of inflammatory signals.
- high-fat diet (HFD) /
- mouse /
- mammary gland development /
- proliferation /
- inflammation /
- protein expression

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图 1 高脂日粮对初情期小鼠采食量(A)、能量摄入(B)、体质量(C)和体质量增加(D)的影响

*、**分别表示与对照组相比差异达0.05、0.01的显著水平(t检验)

Figure 1. Effects of high-fat diet on the feed intake (A), energy intake (B), body weight (C) and body weight gain (D) of pubertal mice

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图 2 高脂日粮对初情期小鼠乳腺形态和导管发育的影响

A、B分别为whole-mount染色后对照组和高脂日粮组小鼠乳腺组织的低倍显微镜照片，C、D分别为whole-mount染色后对照组和高脂日粮组小鼠乳腺组织的高倍显微镜照片；E、F分别为HE染色后对照组和高脂日粮组小鼠乳腺组织的高倍显微镜照片。

Figure 2. Effects of high-fat diet on mammary gland morphology and duct development of pubertal mice

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图 3 高脂日粮对初情期小鼠乳腺增殖及炎症相关蛋白表达的影响

A：Western blot条带；B：图A的蛋白统计图，IGF-I和TLR4的相对表达量计算均以β-actin为内参，*、**分别表示与对照组相比差异达0.05、0.01的显著水平(t检验)。

Figure 3. Effects of high-fat diet on the expressions of proteins associated with proliferation and inflammation in mammary glands of pubertal mice

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表 1 对照组和高脂日粮组日粮成分对比

Table 1 Comparison of dietary composition for control group and high-fat diet group

成分	日粮质量/kg		日粮能量/kJ
成分	对照组	高脂日粮组	对照组	高脂日粮组
酷蛋白(80 Mesh)	189.56	258.51	3 172.56	4 325.38
L-胱氨酸	2.84	3.88	47.57	64.89
玉米淀粉	298.56	0	4 996.78	0
麦芽糊精10	33.17	161.57	555.22	2 703.37
蔗糖	331.74	88.67	5 551.96	1487.91
纤维素BW200	47.39	64.63	0	0
豆油	23.70	32.31	892.28	1 216.50
猪油	18.96	316.68	713.83	11 921.85
复合矿物质S10026	9.48	12.93	0	0
磷酸氢钙	12.32	16.80	0	0
碳酸钙	5.21	7.11	0	0
柠檬酸钾(1 H₂O)	15.64	21.33	0	0
复合维生素V10001	9.48	12.93	158.62	216.27
重酒石酸氢胆碱	1.90	2.59	0	0
FD & C蓝色染料#1	0.05	0.06	0	0
总计	1 000.00	1 000.00	16 088.82	21 936.17

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表 2 高脂日粮对初情期小鼠乳腺(第4对)质量和终末乳芽数量的影响¹⁾

Table 2 Effects of high-fat diet on mammary gland (the fourth pair) weight and the number of terminal end bud of pubertal mice

组别	乳腺质量/g	乳腺质量:体质量	TEB
对照组	0.206±0.006	0.53±0.000 3	81±2.41
高脂日粮组	0.357±0.037^**	0.97±0.001 3^**	29±2.42^**
1) **表示与对照组相比差异达到0.01的显著水平(t检验)

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