Vishnyakova Polina, Artemova Daria, Elchaninov Andrey, Efendieva Zulfiia, Apolikhina Inna, Sukhikh Gennady, Fatkhudinov Timur
National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of Ministry of Healthcare of Russian Federation, Moscow, Russia.
Peoples' Friendship University of Russia (RUDN University), Moscow, Russia.
PeerJ. 2020 Nov 30;8:e10415. doi: 10.7717/peerj.10415. eCollection 2020.
Platelet-rich plasma (PRP), which represents a valuable source of growth factors, is increasingly being applied in regenerative medicine. Recent findings suggest the feasibility of using PRP in the treatment of infertility secondary to refractory thin endometrium. Mesenchymal stem/stromal cells (MSCs) of the endometrium are an essential cellular component responsible for extracellular matrix remodeling, angiogenesis, cell-to-cell communication, and postmenstrual tissue repair. Using a rat model, we examine the effects of autologous PRP on MSCs isolated from the uterus and compare them with the effects of autologous ordinary plasma (OP) and complete growth medium.
MSCs were isolated from uterine tissues via enzymatic disaggregation. Flow cytometry immunophenotyping of the primary cell cultures was complemented by immunocytochemistry for Ki-67 and vimentin. The ability of MSCs to differentiate in osteo-, chondro-, and adipogenic directions was assessed using differentiation-inducing media. The levels of autophagy and apoptosis markers, as well as the levels of matrix metalloproteinase 9 (MMP9) and estrogen receptor α, were assessed by western blotting.
After 24 h incubation, the proliferation index of the PRP-treated MSC cultures was significantly higher than that of the MSC cultures treated with complete growth medium. PRP treatment elevated production of LC3B protein, an autophagy marker, while OP treatment upregulated the expression of stress-induced protein p53 and extracellular enzyme MMP9. The results indicate practical relevance and validity for PRP use in the treatment of infertility.
富含血小板血浆（PRP）是生长因子的宝贵来源，在再生医学中的应用越来越广泛。最近的研究结果表明，使用PRP治疗难治性薄子宫内膜继发的不孕症具有可行性。子宫内膜间充质干/基质细胞（MSCs）是负责细胞外基质重塑、血管生成、细胞间通讯和月经后组织修复的重要细胞成分。我们使用大鼠模型，研究自体PRP对从子宫分离的MSCs的影响，并将其与自体普通血浆（OP）和完全生长培养基的影响进行比较。
通过酶解从子宫组织中分离MSCs。原代细胞培养物的流式细胞术免疫表型分析通过Ki-67和波形蛋白的免疫细胞化学进行补充。使用诱导分化培养基评估MSCs在成骨、成软骨和成脂方向分化的能力。通过蛋白质印迹法评估自噬和凋亡标志物的水平，以及基质金属蛋白酶9（MMP9）和雌激素受体α的水平。
孵育24小时后，PRP处理的MSC培养物的增殖指数显著高于用完全生长培养基处理的MSC培养物。PRP处理提高了自噬标志物LC3B蛋白的产生，而OP处理上调了应激诱导蛋白p53和细胞外酶MMP9的表达。结果表明PRP在不孕症治疗中的实际相关性和有效性。
