利用牛子宫内膜上皮细胞和小鼠子宫内膜炎模型评价胎盘组织源脂质体载药复合物的效果

doi:10.11843/j.issn.0366-6964.2025.10.043

摘要/Abstract

摘要：

本研究旨在评估胎盘来源的磷脂作为抗菌药物载体的可行性，并探索用于治疗子宫内膜炎。用大肠杆菌(E. coli)诱导牛子宫内膜上皮细胞构建体外炎症模型和小鼠子宫内膜炎在体模型，评价庆大霉素-胎盘脂质体复合物(L-GEN)治疗效果。结果显示：用薄膜分散法制备庆大霉素脂质体(L-GEN)；L-GEN混悬液呈乳白色，泛蓝光，且流动性较好；纳米粒度电位仪测得L-GEN粒径为293.7 nm±2.4 nm，分散系数为0.21±0.02。测试发现30 d内L-GEN粒径无显著变化，表明脂质体较稳定。转录组测序分析显示，L-GEN组相比于对照组，对牛子宫内膜细胞影响更小，经KEGG通路富集发现，主要富集在PPAR信号通路影响细胞脂质代谢，降低细胞损伤和炎性，抗菌活性增强。在体试验发现，L-GEN可通过缓解E. coli灌注后小鼠子宫组织的充血肿胀以及炎性细胞浸润，炎性相关基因IL-2、IL-6、IL-1β、TNF-α、TLR2表达水平下降(P≤0.05)，缓解小鼠子宫内膜损伤和炎症反应。经子宫内膜炎细胞模型和小鼠子宫内膜炎在体模型验证，发现胎盘源脂质体载药复合体可有效缓减子宫内膜炎性反应，增强治疗效果，为后续使用组织源脂质体运载抗生素治疗子宫内膜炎提供参考。

关键词: 庆大霉素脂质复合物, 大肠杆菌, 子宫内膜细胞, 子宫内膜炎

Abstract:

This study aimed to evaluate the feasibility of placenta-derived phospholipids as carriers for antibacterial drugs and explore their application in the treatment of endometritis. An in vitroinflammatory model of bovine endometrial epithelial cells and an in vivomodel of endometritis in mice were established using Escherichia coli (E. coli), and the therapeutic effect of gentamicin-placenta lipid complex (L-GEN) was evaluated. Results were as follows: Gentamicin liposomes (L-GEN) were prepared by the film dispersion method; the L-GEN suspension was milky white with a bluish tinge and good fluidity. The particle size of L-GEN was measured by a nanoparticle size and zeta potential analyzer to be 293.7 nm±2.4 nm, with a polydispersity index of 0.21±0.02. No significant change in the particle size of L-GEN was observed within 30 days, indicating the stability of the liposomes. Transcriptome sequencing analysis showed that L-GEN had a smaller impact on bovine endometrial cells compared to the control group. KEGG pathway enrichment analysis revealed that L-GEN mainly enriched in the PPAR signaling pathway, affecting cellular lipid metabolism, reducing cell damage and inflammation, and enhancing antibacterial activity. In vivo experiments found that L-GEN could alleviate the congestion and swelling of the mouse uterus tissue and inflammatory cell infiltration after E. coli perfusion, and the expression levels of inflammatory-related genes IL-2, IL-6, IL-1β, TNF-α, and TLR2 decreased (P≤ 0.05), thereby alleviating endometrial damage and inflammatory response in mice. Through the endometritis cell model and the in vivomodel of endometritis in mice, it was found that the placenta-derived lipid drug delivery complex can effectively alleviate endometrial inflammatory responses and enhance therapeutic effects, providing a reference for the subsequent use of tissue-derived liposomes to carry antibiotics for the treatment of endometritis.

Key words: gentamicin lipid complex, Escherichia coli, endometrial cells, endometritis

中图分类号:

S859.53

周瑾, 庄玉洁, 张雅琳, 谢小雨, 孙明珠, 张雪梅, 王小龙, 张志平, 张君涛. 利用牛子宫内膜上皮细胞和小鼠子宫内膜炎模型评价胎盘组织源脂质体载药复合物的效果[J]. 畜牧兽医学报, 2025, 56(10): 5239-5250.

ZHOU Jin, ZHUANG Yujie, ZHANG Yalin, XIE Xiaoyu, SUN Mingzhu, ZHANG Xuemei, WANG Xiaolong, ZHANG Zhiping, ZHANG Juntao. Antibacterial Test of Placental Tissue-derived Liposome Drug Complex in Endometritis Model[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(10): 5239-5250.

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参考文献 19

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引物Primer	产物大小/bp Product size	序列(5′→3′)Sequence
TNF-α	133	F: CTTCTCATTCCTGCTTGTG R: ACTTGGTGGTTTGCTACG
IL-1β	139	F: CCTGGGCTGTCCTGATGAGAG R: TCCACGGGAAAGACACAGGTA
β-actin	102	F: CCTGCGGCATTCACGAAAC R: GGGGGCGCGATGATCTTG
IL-2	125	F: TTGGACCTCTGCGGCATGTT R: CTCAAATGTGTTGTCAGAGCCC
IL-6	132	F: GGCGGATCGGATGTTGTGAT R: GGACCCCAGACAATCGGTTG
TLR2	113	F: TTTGCTCCTGCGAACTCC R: GCCACGCCCACATCATTC
CD36	120	F: GTGTAACTGTCCTCTGTGTCTAAGC R: GCCTCAAGTGCTGGGTCAAATC
PLIN2	127	F: GATGCTGTGACGACTACTGTGAC R: TGCCACTGACCACGGACTTG
SCD	112	F: CTCCGCTCCGCAGTCTCAG R: TGGTGGTGGTGGTTGTGTAGG
HMGCS1	128	F-CACTACAGCCGAGCATATTCCAAG R-AAGTCCTCCACCTCACAGAAGTATC
ANGPTL4	81	F-TCTGGTGGCTGGTGGTTTGG R-CTGCCGCTGCCTTGGAATG

差异表达基因集名称 Differentially expressed gene set name	差异表达基因数目 Number of differentially expressed genes	上调基因数目 Up-regulate the number of genes	下调基因数目 Number of down-regulated genes
E.coli_vs_GEN	4 344	2 093	2 251
E.coli_vs_L	3 476	1 595	1 881
E.coli_vs_L-GEN	3 268	1 517	1 751
GEN vs_L-GEN	70	44	26
GEN_vs_L	101	61	40

基因名称 Gene name	对照组 Control FPKM	L-GEN组 Treat FPKM	对数 log₂ (Treat /Control)	P值 P-value (Control-vs Treat)
CD36	0.156 739	0.497 800	2.196 005 757 505 25	1.09×10^-6
PLIN2	46.520 181	113.105 376	1.244 822 273 767 47	1.27×10^-42
SCD	19.684 074	11.821 800	1.192 551 399 957 22	4.64×10^-26
HMGCS1	21.208 651	16.174 757	1.048 692 230 259 87	4.65×10^-13
ANGPTL4	1.229 391	13.984 678	3.260 825 231 994 90	2.91×10^-45

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