高脂饮食孕鼠胎盘微环境特征及其意义

doi:10.3969/j.issn.1674-8115.2023.04.001

上海交通大学学报（医学版） ›› 2023, Vol. 43 ›› Issue (4): 397-405.doi: 10.3969/j.issn.1674-8115.2023.04.001

• 论著 · 基础研究 •

高脂饮食孕鼠胎盘微环境特征及其意义

徐一丹¹(), 张骞仁¹, 卢星宇¹, 董艳¹^,²()

^1.上海交通大学医学院附属新华医院内分泌科，上海 200092
^2.上海市儿科医学研究所，上海 200092

收稿日期:2023-02-14 接受日期:2023-03-09 出版日期:2023-04-28 发布日期:2023-04-28
通讯作者: 董艳 E-mail:13862305195@163.com;dongyansh@sjtu.edu.cn
作者简介:徐一丹（1997—），女，硕士生；电子信箱：13862305195@163.com。
基金资助:
国家重点研发计划(2018YFC1004604);上海市科学技术委员会科研基金资助项目(18ZR1431100)

Effect of maternal high-fat diet on placental phenotype in mice

XU Yidan¹(), ZHANG Qianren¹, LU Xingyu¹, DONG Yan¹^,²()

^1.Department of Endocrinology, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
^2.Shanghai Institute of Pediatrics, Shanghai 200092, China

Received:2023-02-14 Accepted:2023-03-09 Online:2023-04-28 Published:2023-04-28
Contact: DONG Yan E-mail:13862305195@163.com;dongyansh@sjtu.edu.cn
Supported by:
National Key Research and Development Program(2018YFC1004604);Scientific Reasearch Fund of Shanghai Science and Technology Commission(18ZR1431100)

摘要/Abstract

摘要：

目的·通过分析高脂饮食对孕鼠胎盘微环境的影响，探讨胎盘微环境改变在母子代际传递中的作用。方法·3周龄C57BL/6J雌性小鼠在适应环境1周后随机分为2组，分别给予高脂饲料（高脂饮食组，HFD组）和普通饲料（对照饮食组，CD组），5周后分别与正常饲喂雄鼠交配怀孕，2组孕鼠按照原饲料继续喂养至孕20 d。于孕20 d时收集母鼠胎盘、肝脏和胎鼠肝脏组织，通过苏木精-伊红染色（hematoxylin-eosin staining，H-E染色）、免疫组织化学（免疫组化）染色、Western blotting以及反转录聚合酶链反应（RT-PCR）等方法观察母鼠高脂饮食对胎盘炎症状态和胎盘结构的影响；检测胎鼠肝脏组织脂质沉积水平；检测子鼠3周龄断乳时的体质量、空腹血糖和腹腔注射葡萄糖糖耐量检测水平。结果·经孕前与孕期饮食干预后，HFD组母鼠体质量增加，肝脏三酰甘油（triacylglycerol，TAG）和总胆固醇（total cholesterol，TC）水平均明显高于CD组母鼠，差异有统计学意义（均P<0.05）。与CD组相比，HFD组胎鼠肝组织可见大小不等的脂性空泡，肝细胞出现脂肪变性，肝脏TAG水平上升，但差异无统计学意义（P>0.05）；HFD组子鼠3周龄时体质量明显升高，空腹血糖水平升高，葡萄糖耐量试验曲线下面积增加（均P<0.05）；免疫组化结果显示HFD组母鼠胎盘内白细胞介素-6（interleukin-6，IL-6）、肿瘤坏死因子α（tumor necrosis factor-α，TNF-α）水平明显增加（均P<0.05），而IL-1β水平无明显变化；H-E染色显示HFD组胎盘迷路区（母胎交换区）面积占比明显减少，通过ImageJ软件统计2组差异具有统计学意义（P<0.05），并且血管间膜增厚，母体血窦狭小；RT-PCR结果显示HFD组胎盘紧密连接相关蛋白Zo-1（zonula occludens 1）和claudin的表达增加，差异有统计学意义（均P<0.05）。结论·母代高脂饮食可引起胎盘炎症因子水平增加和胎盘结构异常，这些改变可能与子代糖脂代谢紊乱有关。

关键词: 高脂饮食, 胎盘, 滋养层细胞, 母子代际传递

Abstract:

Objective ·To analyze the effects of maternal high-fat diet on placental phenotype, and investigate the role of placental microenvironment in the intergenerational transmission. Methods ·The 3-week-old C57BL/6J female mice were fed with either the high-fat diet (HFD group) or the control diet (CD group) for 5 weeks before mating and throughout gestation. Placentas and fetal liver tissues were collected from maternal mice after 20 d of gestation. The effects of maternal HFD on the placental inflammation and placental structure were investigated by hematoxylin-eosin staining (H-E staining), immunohistochemistry, Western blotting and RT-PCR. The lipid deposition levels in fetal livers were also detected. Body weight changes, fasting blood glucose and glucose tolerance levels of the 3-week-old weaned mice were also detected. Results ·The body weight of female mice in the HFD group increased significantly, and the liver triacylglycerol (TAG) and total cholesterol (TC) levels were higher than those in the CD group (all P<0.05). Compared to the CD group, the liver cells of fetus in the HFD group showed steatosis, lipid vacuoles of different sizes, and the content of TAG in the fetal livers in the HFD group increased, but the difference was not statistically significant (P>0.05). Compared to the CD group, the body weight, fasting blood glucose level and the area under the glucose tolerance curve of the 3-week-old weaned mice of the HFD group were increased significantly (all P<0.05). The immunohistochemical results showed that the levels of interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in the placenta in the HFD group were significantly increased (both P<0.05), and the level of IL-1β did not change significantly. In addition, H-E staining of placentas showed that the area proportion of the labyrinth zone (the maternal-fetal exchange area) in the HFD group decreased significantly, which was statistically significant by ImageJ software (P<0.05). The intervascular membrane thickened, and the maternal blood sinuses were narrow. RT-PCR results showed the expressions of placental tight-junction-related protein Zo-1 (zonula occludens 1) and claudin were increased (both P<0.05). Conclusion ·Maternal high-fat diet may result in placental inflammation and abnormal structure, which may be related to glucose and lipid metabolism disorder in offspring.

Key words: maternal high-fat diet, placenta, trophoblast cell, intergenerational transmission

中图分类号:

R715.3

徐一丹, 张骞仁, 卢星宇, 董艳. 高脂饮食孕鼠胎盘微环境特征及其意义[J]. 上海交通大学学报（医学版）, 2023, 43(4): 397-405.

XU Yidan, ZHANG Qianren, LU Xingyu, DONG Yan. Effect of maternal high-fat diet on placental phenotype in mice[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2023, 43(4): 397-405.

图/表 5

图1 2组母鼠体质量、肝脏脂质含量以及摄食量改变Note： A. Weight change curves of female mice before pregnancy. B. TC content in livers of female mice in the two groups. C. TAG content in livers of female mice in the two groups. D. Changes of weekly food intake before pregnancy of female mice in the two groups. ①P=0.015, ②P=0.005, ③P=0.001, ④P=0.009.

Fig 1 Changes in body weight, liver lipid content and food intake of female mice in the two groups

图2 2组母鼠胎盘炎症因子表达水平Note： A. Immunohistochemical staining of IL-6 in placentas and its average optical density. B. Immunohistochemical staining of IL-1β in placentas and its average optical density. C. Immunohistochemical staining of TNF-α in placentas and its average optical density. ①P=0.038, ②P=0.002.

Fig 2 Expression levels of inflammatory cytokines in placentas of female mice in the two groups

图3 2组母鼠胎盘结构改变Note：A. H-E staining sections of the placentas in the two groups. Dotted lines divided different placental zones. JZ—junction zone; LZ—labyrinth zone. B. ImageJ calculated the area proportion of placental labyrinth zone. C. H-E staining sections of the placentas were taken to observe the structure of LZ of the placentas in the two groups (* representing maternal blood sinuses, ▲ representing trophoblast cells, →← representing intervascular membranes). ①P=0.009.

Fig 3 Changes of placental structure in the two groups

图4 2组胎盘紧密连接相关蛋白表达水平Note：A. Placental immunohistochemistry revealed the expression level of ZO-1 in the placentas. B. The expression level of Zo-1 mRNA in placentas was detected by RT-PCR. C. The mRNA expression levels of occludin and claudins in placentas in the two groups were detected by RT-PCR. ①P=0.006, ②P=0.001, ③P=0.012.

Fig 4 Expression levels of tight junction-related proteins in the placentas in the two groups

图5 CD组与HFD组子代表型改变Note： A. H-E staining and oil red staining of fetal liver tissues in the CD group and the HFD group. B. TAG levels in fetal liver tissues of the CD group and the HFD group. C. TC levels in fetal liver tissues of the CD group and the HFD group. D. Body weight of the 3-week-old offsprings in the CD group and the HFD group. E. FBG of the 3-week-old offsprings in the CD group and the HFD group. F/G. Intraperitoneal glucose tolerance test (IPGTT, F) and area under the curve (G) of the 3-week-old offsprings in the CD group and the HFD group. FBG—fasting blood glucose. ①P=0.000, ②P=0.046, ③P=0.009.

Fig 5 Phenotypic changes of offsprings in the CD group and the HFD group

参考文献 21

1	HOFFMAN D J, REYNOLDS R M, HARDY D B. Developmental origins of health and disease: current knowledge and potential mechanisms[J]. Nutr Rev, 2017, 75(12): 951-970.
2	ALFARADHI M Z, OZANNE S E. Developmental programming in response to maternal overnutrition[J]. Front Genet, 2011, 2: 27.
3	PANERA N, MANDATO C, CRUDELE A, et al. Genetics, epigenetics and transgenerational transmission of obesity in children[J]. Front Endocrinol (Lausanne), 2022, 13: 1006008.
4	CAO B G, LIU C X, ZHANG Q R, et al. Maternal high-fat diet leads to non-alcoholic fatty liver disease through upregulating hepatic SCD1 expression in neonate rats[J]. Front Nutr, 2020, 7: 581723.
5	GUDE N M, ROBERTS C T, KALIONIS B, et al. Growth and function of the normal human placenta[J]. Thromb Res, 2004, 114(5/6): 397-407.
6	MESTAN K, YU Y X, MATOBA N, et al. Placental inflammatory response is associated with poor neonatal growth: preterm birth cohort study[J]. Pediatrics, 2010, 125(4): e891-e898.
7	BURTON G J, FOWDEN A L, THORNBURG K L. Placental origins of chronic disease[J]. Physiol Rev, 2016, 96(4): 1509-1565.
8	BARKER D J P. The fetal and infant origins of disease[J]. Eur J Clin Investig, 1995, 25(7): 457-463.
9	HUANG Y H, YE T T, LIU C X, et al. Maternal high-fat diet during pregnancy and lactation affects hepatic lipid metabolism in early life of offspring rat[J]. J Biosci, 2017, 42(2): 311-319.
10	ZHANG Q, YE L, XIN F, et al. Milk fat globule membrane supplementation during suckling ameliorates maternal high fat diet-induced hepatic steatosis in adult male offspring of mice[J]. J Nutr, 2021, 151(6): 1487-1496.
11	GODFREY K M, REYNOLDS R M, PRESCOTT S L, et al. Influence of maternal obesity on the long-term health of offspring[J]. Lancet Diabetes Endocrinol, 2017, 5(1): 53-64.
12	MALTEPE E, FISHER S J. Placenta: the forgotten organ[J]. Annu Rev Cell Dev Biol, 2015, 31: 523-552.
13	ROCHA V Z, LIBBY P. Obesity, inflammation, and atherosclerosis[J]. Nat Rev Cardiol, 2009, 6(6): 399-409.
14	CHALLIER J C, BASU S, BINTEIN T, et al. Obesity in pregnancy stimulates macrophage accumulation and inflammation in the placenta[J]. Placenta, 2008, 29(3): 274-281.
15	陈镇燕, 王琪, 黄光英. 鼠类胎盘结构、血液循环及其来源[J]. 解剖学杂志, 2010, 33(2): 256-259.
	CHEN Z Y, WANG Q, HUANG G Y. Placenta structure, blood circulation and its source in rodents[J]. Chinese Journal of Anatomy, 2010, 33(2): 256-259.
16	郑婉珊, 胡晓倩, 王雁玲, 等. 胎盘屏障建立与维持的机制[J]. 生理学报, 2020, 72(1): 115-124.
	ZHENG W S, HU X Q, WANG Y L, et al. Mechanism for establishment of the placental defensive barrier[J]. Acta Physiologica Sinica, 2020, 72(1): 115-124.
17	FURUKAWA S, TSUJI N, SUGIYAMA A. Morphology and physiology of rat placenta for toxicological evaluation[J]. J Toxicol Pathol, 2019, 32(1): 1-17.
18	WANG Y W, YU H R, TIAO M M, et al. Maternal obesity related to high fat diet induces placenta remodeling and gut microbiome shaping that are responsible for fetal liver lipid dysmetabolism[J]. Front Nutr, 2021, 8: 736944.
19	KRETSCHMER T, TURNWALD E M, JANOSCHEK R, et al. Maternal high fat diet-induced obesity affects trophoblast differentiation and placental function in mice[J]. Biol Reprod, 2020, 103(6): 1260-1274.
20	MARZIONI D, BANITA M, FELICI A, et al. Expression of ZO-1 and occludin in normal human placenta and in hydatidiform moles[J]. Mol Hum Reprod, 2001, 7(3): 279-285.
21	OTANI T, FURUSE M. Tight junction structure and function revisited[J]. Trends Cell Biol, 2020, 30(10): 805-817.

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[2]	徐文晖, 卞京, 郑磊贞. 结直肠癌和高脂饮食关系的研究进展及其防治对策[J]. 上海交通大学学报(医学版), 2021, 41(11): 1514-1517.
[3]	郭怡琼，吴琼，吴雅婷，高璐璐，杨建军. 枸杞多糖和有氧运动对大鼠非酒精性脂肪肝的干预效果及其机制研究[J]. 上海交通大学学报（医学版）, 2020, 40(1): 30-.
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[5]	孙宝航行 1，鲁奕 1，王怡沁 1，周嘉乐 1，林梦璐 1，宋玮 2，姜萌 1，卜军 1. 单个细胞来源胎盘间充质干细胞培养方法的比较研究[J]. 上海交通大学学报（医学版）, 2018, 38(9): 1033-.
[6]	彭海燕，李华萍. 高糖环境对滋养层细胞腺苷单磷酸活化蛋白激酶 α1 亚基表达及增殖的影响[J]. 上海交通大学学报（医学版）, 2018, 38(3): 299-.
[7]	李阳，肖丽，耿爱文，等. 不同饮食结构构建小鼠非酒精性脂肪性肝炎模型的实验研究[J]. 上海交通大学学报（医学版）, 2015, 35(11): 1682-.
[8]	张文天，连锋，薛松. 胎盘生长因子在心肌梗死后的心肌再生治疗中的研究进展[J]. 上海交通大学学报（医学版）, 2014, 34(10): 1543-.
[9]	陈轩，李华萍. 高脂饮食对孕鼠代谢及胎盘炎症和氧化应激反应的影响[J]. 上海交通大学学报（医学版）, 2013, 33(8): 1079-.
[10]	李黎, 徐亮, 滕银成. C型凝集素受体和Toll样受体在子前期重度患者胎盘中的表达[J]. , 2011, 31(8): 1145-.
[11]	翟华玲, 吴晖, 张岚, 等. 雄激素和高脂饮食对雌性大鼠卵巢、垂体、肾上腺超微结构的影响[J]. , 2011, 31(10): 1393-.
[12]	杨科峰, 房玥晖, 张雄, 等. 植物甾醇对大鼠脂质代谢紊乱的预防作用[J]. , 2010, 30(1): 13-.
[13]	熊瑛, 杨祖菁. 母体血浆胎盘亮氨酸氨肽酶和白介素-6与自发性早产的关系[J]. , 2009, 29(8): 955-.

高脂饮食孕鼠胎盘微环境特征及其意义

Effect of maternal high-fat diet on placental phenotype in mice

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