高脂饮食喂养致孕早期小鼠子宫内膜蜕膜化反应损伤。
Impairment of endometrial decidual reaction in early pregnant mice fed with high fat diet.
出版信息
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2022 Apr 25;51(2):204-214. doi: 10.3724/zdxbyxb-2021-0354.
OBJECTIVE
To investigate the effect of obesity induced by high fat diet on decidual reaction of endometrium in mice, and the effect of high fat treatment on decidual reaction of endometrial stromal cells.
METHODS
Twelve 4-week-old healthy C57BL/6J female mice were randomly divided into high fat diet group and control group with 6 mice in each group. They were fed with high fat diet (22 kJ/g) or normal diet (16 kJ/g) for 12 weeks, respectively. The body weight of mice was measured every week. After feeding for 12 weeks, the body length and width of mice were measured, and the levels of fasting serum triglyceride and total cholesterol were determined. Then the mice were mated with healthy C57BL/6J male mice, and the uterine tissues were collected on the seventh day of pregnancy. The decidual cells and collagen fibers in mouse endometrium was observed by HE staining and Masson staining respectively. The expression of decidual reaction related proteins in mouse endometrium were detected by immunohistochemistry and Western blotting. Mouse endometrial stromal cells (mESCs) were isolated and treated with the oleic acid and palmitic acid , and the decidual reaction was induced with estradiol and progesterone. The accumulation of lipid droplets in mESCs was observed by oil red O and Bodipy staining. The cytoskeleton of mESCs was observed by phalloidin staining. The levels of decidual reaction related genes and proteins were detected by real-time fluorescence quantitative PCR and Western blotting.
RESULTS
After feeding for 12 weeks, the body weight of mice in the high fat group was significantly higher than that in the control group ( <0.01), and there was no significant difference in body length between two groups ( >0.05), but the body width of mice in the high fat group was significantly larger than that in the control group ( <0.01), and the levels of serum triglyceride and total cholesterol were significantly higher than those in the control group (Both <0.05). The number of embryo implantation in the high fat group was significantly less than that in the control group ( <0.01). The differentiation of mESCs to decidual cells in high fat group was slow and abnormal. The expression levels of decidual reaction markers bone morphogenetic protein (BMP)2 and homeobox A10 (HOXA10) were lower than those in the control group, and there was significant difference in the expression level of HOXA10 ( <0.01). The results of oil red O and Bodipy staining in mESCs showed that after high fat treatment, the accumulation of lipid droplets increased significantly, phalloidin staining showed abnormal cytoskeleton morphology. The expression levels of decidual reaction related genes , and proteins BMP2, HOXA10 and cyclooxygenase (COX)2 were significantly lower than those in the control group ( <0.05).
CONCLUSION
Obesity induced by high fat diet and high fat treatment can impair the decidual reaction of endometrium and endometrial stromal cells in mice.
目的
探讨高脂饮食诱导的肥胖对小鼠子宫内膜蜕膜反应的影响，以及高脂处理对子宫内膜基质细胞蜕膜反应的影响。
方法
将 12 只 4 周龄健康 C57BL/6J 雌性小鼠随机分为高脂饮食组和对照组，每组 6 只。分别给予高脂饮食（22 kJ/g）或正常饮食（16 kJ/g）喂养 12 周。每周测量小鼠体重。喂养 12 周后，测量小鼠体长和体宽，测定空腹血清甘油三酯和总胆固醇水平。然后将小鼠与健康 C57BL/6J 雄性小鼠交配，在妊娠第 7 天收集子宫组织。分别通过 HE 染色和 Masson 染色观察小鼠子宫内膜中的蜕膜细胞和胶原纤维，通过免疫组化和 Western blot 检测小鼠子宫内膜中蜕膜反应相关蛋白的表达。分离并培养小鼠子宫内膜基质细胞（mESCs），用油酸和棕榈酸处理，用雌二醇和孕酮诱导蜕膜反应。用油红 O 和 Bodipy 染色观察 mESCs 中脂质滴的积累，用鬼笔环肽染色观察 mESCs 的细胞骨架。通过实时荧光定量 PCR 和 Western blot 检测蜕膜反应相关基因和蛋白的表达。
结果
喂养 12 周后，高脂组小鼠体重明显高于对照组（ <0.01），两组体长无明显差异（ >0.05），但高脂组小鼠体宽明显大于对照组（ <0.01），血清甘油三酯和总胆固醇水平明显高于对照组（均 <0.05）。高脂组胚胎着床数明显少于对照组（ <0.01）。高脂组 mESCs 向蜕膜细胞的分化缓慢且异常。蜕膜反应标志物骨形态发生蛋白 2（BMP）2 和同源盒 A10（HOXA10）的表达水平低于对照组，且 HOXA10 的表达水平差异有统计学意义（ <0.01）。mESCs 的油红 O 和 Bodipy 染色结果显示，高脂处理后脂质滴积累明显增加，鬼笔环肽染色显示细胞骨架形态异常。蜕膜反应相关基因和蛋白 BMP2、HOXA10 和环氧化酶（COX）2 的表达水平明显低于对照组（ <0.05）。
结论
高脂饮食诱导的肥胖和高脂处理均可损害小鼠子宫内膜和子宫内膜基质细胞的蜕膜反应。
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