Evaluation of the Effect of Progesterone Packaging in Yak (Bos grunniens) Uterine Fluid Exosomes and Its Effect on Mouse (Mus musculus) Uterine Receptivity
HE Yong-Gang1, YU Tian-Jun2, HE Yu-Long3, WANG Xin-Qiang3, LI Zhi-Jie1, WANG Jing-Lei1, WANG Meng1, LI Tian-Hao1, PAN Yang-Yang1,4*, XU Geng-Quan1,4
1 College of Animal Medicine, Gansu Agricultural University, Lanzhou 730070, China; 2 Animal Husbandry and Veterinary Technical Service Center of Su Bei Mongol Autonomous County, Jiuquan 736300, China; 3 Jiuquan Animal Husbandry and Veterinary General Station, Jiuquan 736300, China; 4 Gansu Provincial Bovine and Sheep Embryo Engineering Technology Research and Development Center, Lanzhou 730070, China
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.
何永刚, 于天军, 何玉龙, 王新强, 李志杰, 王靖雷, 王萌, 李天昊, 潘阳阳, 徐庚全. 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[J]. 农业生物技术学报, 2025, 33(6): 1290-1300.
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. Evaluation of the Effect of Progesterone Packaging in Yak (Bos grunniens) Uterine Fluid Exosomes and Its Effect on Mouse (Mus musculus) Uterine Receptivity. 农业生物技术学报, 2025, 33(6): 1290-1300.
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