Кurinnyi D A, Rushkovsky S R, Demchenko O M, Pilinska M A
State institution «National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine», Melnykova str., 53, Kyiv, 04050, Ukraine.
Educational and Research Center «Institute of Biology and Medicine» of Kyiv Taras Shevchenko National University, Volodymyrska str., 64/13, Kyiv, 01601, Ukraine.
Probl Radiac Med Radiobiol. 2017 Dec;22:208-215.
To identify the possibility of modification by astaxanthin the level of genome damages induced by gamma quanta in the culture of human peripheral blood lymphocytes exposed in vitro on postsynthetic (G2) phase of the first mitotic cycle.
Peripheral blood lymphocytes from four apparently healthy volunteers 35-51 years old were cultivated using modified micromethod. To obtain genomic damages in G2 phase of the first mitotic cycle the part of cultures was irradiated by γ quanta in dose 1.0 Gy through 46 hours of cultivation. Astaxanthin in final con centration 20 μg/ml was exposed to lymphocytes' cultures before the irradiation. Cytogenetic analysis the uniform ly stained slides of metaphase chromosomes was carried out to determine the frequencies of chromosome and chro matid types of aberrations. Using the method of individual cells electrophoresis (Comet assay) the relative level of DNA damages (Tail Moment index) and the frequency of apoptotic cells with high level of DNA fragmentation were evaluated.
Mean group frequencies of chromosome aberrations after gamma irradiation of lymphocytes in vitro exceed ed those without radiation exposure and were 72.35 ± 1.17 and 2.46 ± 0.30 per 100 metaphases, respectively (p < 0.001), mainly due to chromatid type of aberrations (58.32 ± 1.29 per 100 metaphases). Adding of astaxanthin into culture medium before the irradiation did not result in changes as in the frequency of chromosomal damages (71.54 ± 1.34 per 100 metaphases) as in the spectrum of aberrations - also prevailed chromatid type of aberrations (58.47 ± 1.47 per 100 metaphases). The increase of Tail Moment index after radiation exposure (from 3.84 ± 0.36 to 12.06 ± 1.88, respectively, p < 0.001) and lack of significant impact of astaxanthin on this index in the irradiated lym phocytes (8.96 ± 2.39, p > 0.05) was established, ie astaxanthin didn't change the relative level of radiation induced DNA damages. Also apoptogenic effect of astaxanthin was not found: frequency of apoptotic cells were (2.25 ± 1.49) % in cultures of intact lymphocytes, (2.08 ± 1.54) % in irradiated cultures and (1.78 ± 1.25) % under joint action of gamma radiation and astaxanthin (p > 0.05).
Noimpactofastaxanthinongenomicinstabilityinducedbygammairradiation invitroinculturesof human peripheral blood lymphocytes on postsynthetic (G2) phase of first mitotic cycle had been established.
确定虾青素对处于第一个有丝分裂周期合成后期（G2期）的体外培养的人外周血淋巴细胞中γ量子诱导的基因组损伤水平进行修饰的可能性。
采用改良的微量方法培养来自4名35 - 51岁表面健康志愿者的外周血淋巴细胞。为了在第一个有丝分裂周期的G2期获得基因组损伤，在培养46小时后，用1.0 Gy剂量的γ量子对部分培养物进行照射。在照射前，将终浓度为20μg/ml的虾青素作用于淋巴细胞培养物。进行中期染色体均匀染色玻片的细胞遗传学分析，以确定染色体和染色单体类型畸变的频率。使用单细胞电泳法（彗星试验）评估DNA损伤的相对水平（尾矩指数）以及具有高水平DNA片段化的凋亡细胞频率。
体外γ照射淋巴细胞后，染色体畸变的平均组频率超过未照射组，分别为每100个中期细胞72.35±1.17和2.46±0.30（p<0.001），主要是染色单体类型的畸变（每100个中期细胞58.32±1.29）。在照射前向培养基中添加虾青素，无论是染色体损伤频率（每100个中期细胞71.54±1.34）还是畸变谱均未发生变化，染色单体类型的畸变也占主导（每100个中期细胞58. .47±1.47）。确定了照射后尾矩指数增加（分别从3.84±0.36增加到12.06±1.88，p<0.001），而虾青素对受照射淋巴细胞的该指数无显著影响（8.96±2.39，p>0.05），即虾青素未改变辐射诱导的DNA损伤的相对水平。此外，未发现虾青素的凋亡诱导作用：完整淋巴细胞培养物中凋亡细胞频率为（2.25±1.49）%，照射培养物中为（2.08±1.54）%，γ辐射与虾青素联合作用下为（1.78±1.25）%（p>0.05）。
已确定在第一个有丝分裂周期合成后期（G2期）的人外周血淋巴细胞体外培养中，虾青素对γ辐射诱导的基因组不稳定性无影响。
