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目的:观察蝎毒多肽(SVP)及IL-3对M-CSF依赖细胞株M-NFS-60促增殖作用以及对辐射后MNFS-60细胞增殖的影响。方法:采用AlamarBlue摄入法检测细胞的增殖情况。①选择10个细胞浓度,分别于0 h、2 h、4 h、5.5 h和20 h,检测细胞的增殖情况,确定M-NFS-60细胞对AlamarBlue反应所需的最佳细胞密度和孵育时间;②采用最佳细胞密度和孵育时间,观察10个SVP组分促进M-NFS-60细胞的增殖作用,筛选出SVP的有效组分;③M-NFS-60细胞经60Coγ-射线照射后分别给予生理盐水、SVP、IL-3、SVP+IL-3处理48 h,测定细胞增殖情况。结果:①M-NFS-60细胞的接种密度为5×10~4 cells/mL时,对AlamarBlue的还原能力最强,孵育时间8 h后,M-NFS-60细胞对AlamarBlue的还原率开始增强,72 h达最高;②从10个SVP组分中筛选出Ⅱ3、Ⅳ能明显促进M-NFS-60细胞的增殖;③SVPⅡ3、Ⅲ3、Ⅳ或IL-3处理辐射后的细胞48 h,细胞的增殖率(%)分别为121.58±2.62(Ⅱ3)、123.39±4.45(Ⅲ3)、123.51±5.04(Ⅳ)、140.12±1.68(IL-3),与空白对照组(100.00±0.01)相比差异有显著性(P<0.01);Ⅱ3、Ⅲ3、Ⅳ分别与IL-3联用处理细胞48 h后的增殖率分别为163.98±9.20(Ⅱ3+IL3)、159.89±8.31(Ⅳ+Ⅱ3)、148.92±9.74(Ⅲ3+IL3),与SVP处理组比较差异有显著性(P<0.05)。结论:蝎毒中存在促进M-NFS-60增殖的有效多肽组分,这些多肽组分能明显促进辐射后M-NFS-60细胞的增殖,并与IL-3对辐射后M-NFS-60细胞的增殖具有协同作用。
Objective: To observe the effects of scorpion venom polypeptide (SVP) and IL-3 on the proliferation of M-CSF-dependent M-NFS-60 cells and the proliferation of MNFS-60 cells after irradiation. Methods: The proliferation of cells was detected by AlamarBlue ingestion. ① The cell proliferation was measured at 10 h, 2 h, 4 h, 5.5 h and 20 h, respectively, and the optimal cell density and incubation time for M-NFS-60 cells response to AlamarBlue ; ②Those optimal SVPs were used to observe the proliferation of M-NFS-60 cells by using the optimal cell density and incubation time, and the effective components of SVP were screened out; ③M-NFS-60 cells were irradiated with 60Coγ- Normal saline, SVP, IL-3, SVP + IL-3 for 48 h, the cell proliferation was measured. Results: ①M-NFS-60 cells had the strongest reducing ability to AlamarBlue when the inoculation density was 5 × 10 ~ 4 cells / mL. After 8 h of incubation, the reduction rate of AlamarBlue in M-NFS-60 cells began to increase, 72h, and reached the highest level at 72 hrs. (2) Screening out of the 10 SVP components, Ⅱ3 and Ⅳ could significantly promote the proliferation of M-NFS-60 cells; ③SVPⅡ3, Ⅲ3, Ⅳ or IL-3 treatment of irradiated cells 48 h, cell proliferation (%) Were 121.58 ± 2.62 (Ⅱ 3), 123.39 ± 4.45 (Ⅲ 3), 123.51 ± 5.04 (Ⅳ) and 140.12 ± 1.68 (IL-3) respectively, which were significantly different from those of the blank control group (100.00 ± 0.01) (P <0.01). The proliferation rates of cells treated with IL-3, IL-3 and IL-3 after treated with IL-3 were 163.98 ± 9.20 (Ⅱ3 + IL3), 159.89 ± 8.31 (Ⅳ + Ⅱ3), 148.92 ± 9.74 (Ⅲ 3 + IL3), which showed significant difference compared with SVP group (P <0.05). Conclusion: There are effective polypeptide components in scorpion venom that promote the proliferation of M-NFS-60. These peptide components can significantly promote the proliferation of M-NFS-60 cells after irradiated with IL-3, Cell proliferation has a synergistic effect.