牦牛宫腔液外泌体包装孕酮效果的评价及其对小鼠子宫容受性的影响

doi:10.3969/j.issn.1674-7968.2025.06.010

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Abstract Progesterone (P4) is an important reproductive hormone that regulates the synchronisation of endometrial tolerance and embryonic development, and ensures successful follicle implantation; exosomes have good biocompatibility, low immunogenicity and are excellent carriers for drug transport. In order toexplore whether yak (Bos grunniens) uterine fluid-derived exosomes could be used as delivery vehicles fordrugs such as hormones, proteins or RNAs. In this study, exosomes of yak uterine fluid were obtained by ultra- high-speed centrifugation, and the morphological and marker proteins (CD81 and TSG101) were identified by transmission electron microscopy and Western blot, respectively, and the histological changes of mouse (Musmusculus) organs were analyzed by intraperitoneal injection of mice to evaluate their safety. Yak uterine fluid exosomes were combined with progesterone ultrasonication packaging, and the effect of exosome packaging on progesterone was analyzed by comparing the endometrial thickness and the expression levels of leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) in mice. The results showed that the exosomes of yak uterine fluid obtained by ultracentrifugation were typical in morphology, and the labeled proteins of CD81 and TSG101 were clearly expressed. The expression levels of exosome marker proteins CD81 and TSG101 decreased by sonication, which were 0.74 and 0.83 times higher than those in the control group, respectively. The packaging rate of exosome-packed progesterone was (48.4±1.29)%. Through histological comparison, the optimal concentration of dimethyl sulfoxide (DMSO) as a progesterone co-solvent for exosome packaging was0.5%, and yak exosomes had no damage to mouse organs and tissues. Progesterone after exosome packaging significantly increased endometrial thickness and up-regulated the mRNA and protein expression levels of LIF in mice (P<0.05). The mRNA and protein expression levels of endometrial IL-6 in the single progesterone-treated group were lower than those in the control group, while the expression levels of IL-6 in the exosome- packaged progesterone-treated group increased. The results showed that yak uterine fluid exosomes could be used as effective drug delivery vehicles for progesterone, which could significantly improve progesterone bioavailability. This study provides a theoretical reference for the application of reproductive tract exosomes in the regulation of animal reproduction.

Key words： Yak Exosomes Progesterone Uterine receptivity

Received: 12 September 2024

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Corresponding Authors: *panyangyang_2007@126.com

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	Articles by authors
	HE Yong-Gang
	YU Tian-Jun
	HE Yu-Long
	WANG Xin-Qiang
	LI Zhi-Jie
	WANG Jing-Lei
	WANG Meng
	LI Tian-Hao
	PAN Yang-Yang
	XU Geng-Quan

Cite this article:

HE Yong-Gang,YU Tian-Jun,HE Yu-Long, et al. Evaluation of the Effect of Progesterone Packaging in Yak (Bos grunniens) Uterine Fluid Exosomes and Its Effect on Mouse (Mus musculus) Uterine Receptivity[J]. 农业生物技术学报, 2025, 33(6): 1290-1300.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2025.06.010 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2025/V33/I6/1290

[1] 董峰.2015. 苦马豆素诱导山羊胎盘滋养层细胞凋亡的信号转导通路研究[D]. 博士学位论文, 西北农林科技大学, 导师 : 童德文 , pp. 12-20.
(Dong F.2015. Study on the signal transduction pathway of apoptosis induced by pi-cloram induced goat placental trophoblast cells[D]. The-sis for Ph. D., Northwest Agriculture and Forestry Uni-versity, Supervisor: Tong D W, pp. 12-20.)
[2] 李琦, 秦雪, 冯瑞, 等. 2022. 孕酮对奶牛子宫内膜上皮细胞凋亡和容受性的影响[J]. 中国兽医学报, 42(09): 1915-1922.
(Li Q, Qin X, Feng R, et al.2022. Effect of pro-gesterone on apoptosis and tolerance of endometrial epi-thelial cells in dairy cows[J]. Chinese Journal of Veteri-nary Science, 42(09): 1915-1922.)
[3] 王红战, 陈艳茹, 李琦, 等. 2024. LIF 调控奶牛子宫内膜上皮细胞容受性基因表达研究[J]. 黑龙江动物繁殖, 32(01): 1-6.
(Wang H Z, Chen Y R, Li Q, et al.2024. LIF regulates the expression of tolerance genes in endometri-al epithelial cells of dairy cows[J]. Heilongjiang Animal Breeding, 32(01): 1-6.)
[4] 王丽娜, 孙丰源. 2024. 外泌体在眼科应用的研究进展[J]. 国际眼科杂志 , 24(04): 596-600.
(Wang L N, Sun F Y.2024. Progress of exosomes in ophthalmology[J]. Inter-national Journal of Ophthalmology, 24(04): 596-600.)
[5] 王正刚.2022. EphA2 靶向性载药微囊泡的构建及其抗骨肉瘤作用研究[D]. 博士学位论文, 华中科技大学, 导师: 陈安民 , pp. 85-102.
(Wang Z G.2022. Construction of EphA2-targeted drug-carrying microvesicles and its anti-osteosarcoma effect[D]. Thesis for Ph. D., Huazhong University of Science and Technology, Supervisor: Chen A M, pp. 85-102.)
[6] 文武龙, 张炜烨, 贺雨欣, 等. 2023. 基于牛乳外泌体与脂质体的杂化外泌体的制备及其负载青藤碱治疗类风湿性关节炎的药效学研究[J]. 中国中药杂志 , 49(15): 4069-4077.
(Wen W, Zhang W, He Y, et al.2023. Prepa-ration of hybrid exosomes based on bovine milk exo-somes and liposomes and pharmacodynamic study of their loading with cytisine in the treatment of rheuma-toid arthritis[J]. Chinese Journal of Traditional Chinese Medicine, 49(15): 4069-4077.)
[7] 袁晨丰.2023. 卵泡液外泌体对猪卵泡细胞增殖、类固醇合成、抗氧化应激和趋化因子分泌的影响[D]. 博士学位论文, 吉林大学, 导师: 周虚, pp. 1-8; 17-30.
(Yuan C F.2023. Effects of follicular fluid exosomes on prolifera-tion, steroid synthesis, anti-oxidative stress and chemo-kine secretion in porcine follicular cells[D]. Thesis for Ph.D., Jilin University, Supervisor: Zhou X, pp. 1-8; 17-30.)
[8] 张明, 郑洁, 甘潇, 等. 2013. 孕酮、干扰素 τ 对牛子宫内膜上细胞外基质及其相关配体表达的影响[J]. 畜牧兽医学报 , 44(02): 309-315.
(Zhang M, Zheng J, Gan X, et al.2013. Effects of progesterone and interferon τ on the ex-pression of extracellular matrix and related ligands in bovine endometrium[J]. Journal of Animal Husbandry and Veterinary Science, 44(02): 309-315.)
[9] 赵婷婷, 李俊峰, 张立婷. 2024. 间充质干细胞源性外泌体对肝纤维化潜在治疗机制的研究进展[J]. 中国临床药理学与治疗学, 29(04): 475-480.
(Zhao T T, Li J F, Zhang L T.2024. Progress in the study of potential therapeutic mechanisms of mesenchymal stem cell-derived exo-somes for liver fibrosis[J]. Chinese Clinical Pharmacolo-gy and Therapeutics, 29(04): 475-480.)
[10] 周琦琦, 孙振, 刘洋洋, 等. 2021. 过表达 NAMPT 脐带间充质干细胞外泌体的抗衰老作用[J]. 扬州大学学报(农业与生命科学版), 42(04): 1-6.
(Zhou Q Q, Sun Z, Liu Y Y, et al.2021. Anti-aging effect of exosomes from um-bilical cord mesenchymal stem cells overexpressing NAMPT[J]. Journal of Yangzhou University(Agricultur-al and Life Sciences), 42(04): 1-6.)
[11] Ashary N, Tiwari A, Modi D.2018. Embryo implantation: War in times of love[J]. Endocrinology, 159: 1188-1198.
[12] Blavy P, Friggens N C, Nielsen K R, et al.2018. Estimating probability of insemination success using milk proges-terone measurements[J]. Journal of Dairy Science, 101(2): 1648-1660.)
[13] Canellada A, Alvarez I, Berod L, et al.2008. Estrogen and progesterone regulate the IL-6 signal transduction path-way in antibody secreting cells[J]. Steroids, 111(3-5): 255-261.
[14] Davaran S, Alimirzalu S, Nejati-Koshki K, et al.2014. Physi-cochemical characteristics of Fe3O4 magnetic nanocom-posites based on Poly(N-isopropylacrylamide)for anti-cancer drug delivery[J]. Asian Pacific Journal of Cancer Prevention, 15(1): 49-54.
[15] Feng R, Qin X, Li Q, et al.2022. Progesterone regulates in-flammation and receptivity of cells via the NF-κB and LIF/STAT3 pathways[J]. Theriogenology, 186: 50-59.
[16] Giacomini E, Scotti G M, Vanni V S, et al.2021. Global tran-scriptomic changes occur in uterine fluid-derived extra-cellular vesicles during the endometrial window for em-bryo implantation[J]. Human Reproduction, 36: 2249-2274.
[17] Hajipour H, Farzadi L, Roshangar L, et al.2021. A human chorionic gonadotropin(hCG)delivery platform using engineered uterine exosomes to improve endometrial re-ceptivity[J]. Life Sciences, 275: 119351.
[18] Hajipour H, Ghorbani M, Kahroba H, et al.2019. Arginyl-gly-cyl-aspartic acid(RGD)containing nanostructured lipid carrier co-loaded with doxorubicin and sildenafil citrate enhanced anti-cancer effects and overcomes drug resis-tance[J]. Process Biochemistry, 84: 172-179.
[19] Huppertz B, Kertschanska S, Frank H G, et al.1996. Extracel-lular matrix components of the placental extravillous tro-phoblast: Immunocytochemistry and ultrastructural dis-tribution[J]. Histochemistry and Cell Biology, 106(3): 291-301.
[20] Kalluri R, Lebleu V S.2020. The biology function and bio-medical applications of exosomes[J]. Science, 367:6977-7017.
[21] Meng X, Chen C, Qian J, et al.2023. Energy metabolism and maternal-fetal tolerance working in decidualization[J]. Frontiers in Immunology, 14: 1203719-1203731.
[22] Moench R, Grimmig T, Kannen V, et al.2016. Exclusive inhi-bition of PI3K/Akt/mTOR signaling is not sufficient to prevent PDGF-mediated effects on glycolysis and prolif-eration in colorectal cancer[J]. Oncotarget, 7(42): 68749-68767.
[23] Mullen M P, Elia G, Hilliard M, et al.2012. Proteomic charac-terization of histotroph during the preimplantation phase of the estrous cycle in cattle[J]. Journal of Proteome Re-search, 11: 3004-3018.
[24] Pegtel D M, Gould S J.2019. Exosomes[J]. Annual Review of Biochemistry, 88: 487-514.
[25] Stefani C, Miricescu D,Stanescu-Spinu I I, et al. 2021.
[26] Growth factors, PI3K/AKT/mTOR and MAPK signal-ing pathways in colorectal cancer pathogenesis: Where are we now?[J]. International Journal of Molecular Sci-ences, 22(19): 10260.
[27] Szekeres-Bartho J, Halasz M, Palkovics T.2009. Progesterone in pregnancy: Receptor-ligand interaction and signaling pathways[J]. Journal of Reproductive Immunology, 83(1-2): 60-64.
[28] Teh W T, McBain J, Rogers P.2016. What is the contribution of embryo-endometrial asynchrony to implantation fail-ure?[J]. Journal of Assisted Reproduction and Genetics,33: 1419-1430.
[29] Wollert T, Hurley J H.2010. Molecular mechanism of multi-vesicular body biogenesis by ESCRT complexes[J]. Na-ture, 464(7290): 864-869.
[30] Zhang H.2018. Extracellular vesicles in reproduction-small packages, big consequences[J]. Cell Communication and Signaling, 16(1): 71.
[31] Zhang X, Wei H.2021. Role of decidual natural killer cells in human pregnancy and related pregnancy complications[J]. Frontiers in Immunology, 12: 728291.