Demyashkin Grigory A, Vadyukhin Matvey A, Shekin Vladimir I
I.M. Sechenov First Moscow State Medical University (Sechenov University), Moscow, Russia.
A. Tsyb Medical Radiological Research Centre, Ministry of Health of Russian Federation, Obninsk, Russia.
J Reprod Infertil. 2023 Apr-Jun;24(2):94-100. doi: 10.18502/jri.v24i2.12494.
At present, the damage of male reproductive function caused by electron irradiation, as well as the development of methods for its correction are the relevant topics for further research. In fact, the effect of leukocyte-poor platelet-rich plasma (LP-PRP) growth factors are poorly investigated on different aspects of male fertility.
In this study, Wistar rats were divided into four groups; I) control which were injected with saline; II and III) groups (n=65) whose testes were locally irradiated with electrons to a dose of 2 Gy (linear accelerator "NOVAC-11"); III) the group (n=30) which received LP-PRP for 11 weeks after irradiation; and IV) animals (n=30) which received only LP-PRP (conditional control). The testes were studied by histological, immunohistochemical (IHC), western blotting, and TUNEL methods using Ki-67, Bcl-2, and p53. Comparison between groups was performed and p<0.01 was set as the level of significance.
The results showed a decrease in the expression levels of Ki-67 and Bcl-2 besides an increase in p53-positive cells by the end of the experiment (p<0.01). After injection of LP-PRP, a gradual restoration of the proliferative activity of gametes was noted, which was confirmed by an increase in the proportion of Ki-67- and Bcl-2-positive germ cells (46.4±2.3, p<0.01 and 23.5±1.1, respectively, p<0.01).
Ki-67 expression and TUNEL analysis in the testes revealed a modulation of the proliferative-apoptotic balance towards apoptosis of germ cells after 2 Gy local electron irradiation. A tendency to restore the proliferative-apoptotic balance was noted after LP-PRP injections as indicated by increase in Ki-67-positive germ cells.
目前，电子辐射对男性生殖功能的损害以及其纠正方法的开发是进一步研究的相关课题。事实上，白细胞贫乏的富血小板血浆（LP-PRP）生长因子对男性生育力不同方面的影响研究较少。
在本研究中，将Wistar大鼠分为四组；I）注射生理盐水的对照组；II和III）两组（n = 65），其睾丸局部接受2 Gy电子照射（直线加速器“NOVAC - 11”）；III）照射后接受LP-PRP治疗11周的组（n = 30）；IV）仅接受LP-PRP的动物组（n = 30）（条件对照组）。使用Ki-67、Bcl-2和p53通过组织学、免疫组织化学（IHC）、蛋白质印迹和TUNEL方法对睾丸进行研究。进行组间比较，将p < 0.01设定为显著性水平。
结果显示，到实验结束时，Ki-67和Bcl-2的表达水平降低，p53阳性细胞增加（p < 0.01）。注射LP-PRP后，观察到配子增殖活性逐渐恢复，这通过Ki-67和Bcl-2阳性生殖细胞比例的增加得到证实（分别为46.4±２.3，p < 0.01和23.5±1.1，p < 0.01）。
睾丸中的Ki-67表达和TUNEL分析显示，2 Gy局部电子照射后，增殖-凋亡平衡向生殖细胞凋亡方向调节。LP-PRP注射后，观察到增殖-凋亡平衡有恢复的趋势，表现为Ki-67阳性生殖细胞增加。
