生物复合纳米颗粒PLGA-RES在保护卵母细胞免受玻璃化刺激方面的卓越性能。
Superior performance of biocomposite nanoparticles PLGA-RES in protecting oocytes against vitrification stimuli.
作者信息
Hai Guiping, Bai Jiachen, Liu Yucheng, Li Jun, Liu Aiju, Wang Jingjing, Liu Qian, Liu Weijun, Wan Pengcheng, Fu Xiangwei
机构信息
College of Animal Science, Xinjiang Agricultural University, Ürümqi, China.
State Key Laboratory of Sheep Genetic Improvement and Healthy Breeding, Institute of Animal Husbandry and Veterinary Sciences, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi, China.
出版信息
Front Bioeng Biotechnol. 2024 Mar 11;12:1376205. doi: 10.3389/fbioe.2024.1376205. eCollection 2024.
Irreversible cryogenic damage caused by oocyte vitrification limits its widespread use in female fertility preservation. In recent years, nanoparticles (NPs) have gained great attention as potential alternatives in protecting oocytes against cryoinjuries. In this paper, a novel composite nanoparticle, poly (lactic-co-glycolic acid)-resveratrol (PLGA-RES) was designed to improve the biocompatibility and sustained release properties by encapsulating natural antioxidant RES into PLGA NPs. Firstly, biotoxicity and oxidation resistance of PLGA-RES were determined, and the results showed that PLGA-RES had nontoxic effect on oocyte survival during maturation (IVM) (97.08% ± 0.24% vs. 98.89% ± 1.11%, > 0.05). Notably, PLGA-RES even increased maturation (65.10% ± 4.11% vs. 52.85% ± 2.87%, < 0.05) and blastocyst rate (56.13% ± 1.36% vs. 40.91% ± 5.85%, < 0.05). Moreover, the reduced reactive oxygen species (ROS) level (13.49 ± 2.30 vs. 34.07 ± 3.30, < 0.01), increased glutathione (GSH) (44.13 ± 1.57 vs. 37.62 ± 1.79, < 0.01) and elevated mitochondrial membrane potential (MMP) levels (43.10 ± 1.81 vs. 28.52 ± 1.25, < 0.01) were observed in oocytes treated with PLGA-RES when compared with that of the control group. Subsequently, the role of PLGA-RES played in oocytes during vitrification was systematically evaluated. The results showed that the addition of PLGA-RES during vitrification and thawing significantly improved the survival rate (80.42% ± 1.97% vs. 75.37% ± 1.3%, < 0.05). Meanwhile, increased GSH (15.09 ± 0.86 vs. 14.51 ± 0.78, < 0.01) and mitochondrial membrane potential (22.56 ± 3.15 vs. 6.79 ± 0.60, < 0.01), decreased reactive oxygen species levels (52.11 ± 2.95 vs. 75.41 ± 7.23, < 0.05) and reduced mitochondrial abnormality distribution rate (25.00% ± 0.29% vs. 33.33% ± 1.15%, < 0.01) were assessed in vitrified MII oocytes treated with PLGA-RES. Furthermore, transcriptomic analyses demonstrated that PLGA-RES participated in endocytosis and PI3K/AKT/mTOR pathway regulation, which was verified by the rescued expression of ARRB2 and ULK3 protein after PLGA-RES treatment. In conclusion, PLGA-RES exhibited potent antioxidant activity, and could be used as an efficacious strategy to improve the quality of vitrified oocytes.
卵母细胞玻璃化冷冻所导致的不可逆低温损伤限制了其在女性生育力保存中的广泛应用。近年来，纳米颗粒（NPs）作为保护卵母细胞免受冷冻损伤的潜在替代物受到了广泛关注。本文设计了一种新型复合纳米颗粒聚（乳酸 - 乙醇酸） - 白藜芦醇（PLGA - RES），通过将天然抗氧化剂白藜芦醇（RES）包裹在聚乳酸 - 乙醇酸共聚物（PLGA）纳米颗粒中来提高其生物相容性和缓释性能。首先，测定了PLGA - RES的生物毒性和抗氧化性，结果表明PLGA - RES对卵母细胞体外成熟（IVM）过程中的存活无毒性影响（97.08% ± 0.24% 对 98.89% ± 1.11%，> 0.05）。值得注意的是，PLGA - RES甚至提高了卵母细胞的成熟率（65.10% ± 4.11% 对 52.85% ± 2.87%，< 0.05）和囊胚率（56.13% ± 1.36% 对 40.91% ± 5.85%，< 0.05）。此外，与对照组相比，经PLGA - RES处理的卵母细胞中活性氧（ROS）水平降低（13.49 ± 2.30 对 34.07 ± 3.30，< 0.01），谷胱甘肽（GSH）增加（44.13 ± 1.57 对 37.62 ± 1.79，< 0.01），线粒体膜电位（MMP）水平升高（43.10 ± 1.81 对 28.52 ± 1.25，< 0.01）。随后，系统评估了PLGA - RES在卵母细胞玻璃化冷冻过程中所起的作用。结果表明，在玻璃化冷冻和解冻过程中添加PLGA - RES可显著提高存活率（80.42% ± 1.97% 对 75.37% ± 1.3%，< 0.05）。同时，在经PLGA - RES处理的玻璃化MII期卵母细胞中，GSH增加（15.09 ± 0.86 对 14.51 ± 0.78，< 0.01），线粒体膜电位升高（22.56 ± 3.15 对 6.79 ± 0.60，< 0.01），活性氧水平降低（52.11 ± 2.95 对 75.41 ± 7.23，< 0.05），线粒体异常分布率降低（25.00% ± 0.29% 对 33.33% ± 1.15%，< 0.01）。此外，转录组分析表明PLGA - RES参与内吞作用和PI3K/AKT/mTOR信号通路调控，这在PLGA - RES处理后ARRB2和ULK3蛋白的表达恢复中得到了验证。综上所述，PLGA - RES具有强大的抗氧化活性，可作为提高玻璃化冷冻卵母细胞质量的有效策略。
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