Department of Biophysics, Faculty of Physics, Lomonosov Moscow State University, Moscow, Russian Federation.
Department of Radiobiogy, Institute of theoretical and experimental biophysics, Russian Academy of Sciences (ITEB RAS), Russian Federation.
Biochem Biophys Res Commun. 2024 Nov 12;733:150442. doi: 10.1016/j.bbrc.2024.150442. Epub 2024 Jul 23.
Radioprotectors help to protect the body or at least minimize the negative consequences of radiation exposure. The present study aimed to assess the radioprotective potential of Helianthus tuberosus L. polysaccharide (HTLP) in vitality and micronuclei tests. To assess the cytotoxic effects of HTLP, both vitality and MTT reductase assays were conducted.
RAW 264.7 cells viability was assessed 24 h after adding 200 μg/ml HTLP solution by staining cell cultures with propidium iodide and bis-benzimide to detect the nuclei of dead cells and the total number of cells in culture. To assess cell viability via cellular metabolic activity MTT test was used. In this work outbred 24-30 g 5-months old SHK mice have been used. Irradiation was provided with proton beams with an energy of 660 MeV at a dose rate of 80 Gy with doses 1.5 Gy for micronuclei test and 8.5 Gy for survival test. Whole body X-ray irradiation was conducted using the RUT-15 therapeutic X-ray unit with doses of 1.5 Gy for MN test and 6.5 Gy for survival. The HTLP sterile solution in dose 100 μg/animal was injected into the tail vein 15 min before X-ray or proton irradiation.
s: Vitality test showed no significant differences between the control group and cells treated with 200 μl of 200 μg/ml HTLP solution, though a greater variability was noted. In contrast, the MTT assay indicated enhanced cell viability in the HTLP-treated cells. HTLP does not exert any toxic effects in cell culture. Moreover, results of MTT reductase assay shows, that HTLP may enhance the cells' metabolic activity. Animals pre-treated with HTLP displayed a significant reduction in micronuclei formation, showing five times fewer micronuclei in bone marrow cells compared to the non-treated group. This comparison highlights HTLP's potential protective effect against radiation-induced chromosomal damage. HTLP treatment demonstrates a significant reduction in hazard compared to the control, indicating its protective effects against irradiation. Thus, it can be concluded that the use of HTLP increases the likelihood of animal survival under the ionizing effects of X-rays and protons. The survival analysis reveals that the HTLP-treated groups exhibit a higher survival rate compared to both the control and Cysteamine-treated groups, suggesting a significant protective effect of HTLP against irradiation, regardless of the type of irradiation (proton or X-ray) with p < 0.0001.
辐射防护剂有助于保护身体，或者至少将辐射暴露的负面影响降到最低。本研究旨在评估 Helianthus tuberosus L. 多糖 (HTLP) 在活力和微核试验中的辐射防护潜力。为了评估 HTLP 的细胞毒性作用，进行了活力和 MTT 还原酶测定。
在添加 200μg/ml HTLP 溶液 24 小时后，通过用碘化丙啶和双苯并咪唑染色细胞培养物来评估 RAW 264.7 细胞的活力，以检测死细胞的细胞核和培养物中总细胞数。使用 MTT 试验通过细胞代谢活性来评估细胞活力。在这项工作中，使用了 24-30g 5 个月大的 SHK 小鼠。辐照采用能量为 660MeV 的质子束，剂量率为 80Gy，剂量为 1.5Gy 进行微核试验，8.5Gy 进行存活试验。全身 X 射线照射采用 RUT-15 治疗 X 射线装置进行，剂量为 1.5Gy 进行 MN 试验，6.5Gy 进行存活试验。将无菌 HTLP 溶液以 100μg/动物的剂量在 X 射线或质子辐照前 15 分钟经尾静脉注射。
活力试验显示，对照组和用 200μl 200μg/ml HTLP 溶液处理的细胞之间没有显著差异，但变异更大。相比之下，MTT 测定表明 HTLP 处理的细胞活力增强。HTLP 在细胞培养中没有产生任何毒性作用。此外，MTT 还原酶测定结果表明，HTLP 可能增强细胞的代谢活性。用 HTLP 预处理的动物显示出微核形成的显著减少，与未处理组相比，骨髓细胞中的微核减少了五倍。这一比较突出了 HTLP 对辐射诱导的染色体损伤的潜在保护作用。与对照组相比，HTLP 处理显著降低了危害，表明其对辐照的保护作用。因此，可以得出结论，HTLP 的使用增加了动物在 X 射线和质子电离效应下存活的可能性。生存分析表明，与对照组和半胱胺处理组相比，HTLP 处理组的存活率更高，表明 HTLP 对辐照有显著的保护作用，无论辐照类型（质子或 X 射线）如何，p<0.0001。
