蝎毒多肽有效组分对辐射后M-NFS-60细胞的促增殖作用

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M—NFS-60细胞增殖的影响。方法：采用Alamar Blue摄入法检测细胞的增殖情况。①选择10个细胞浓度．分别于0h、2h、4h、5．5h和20h，检测细胞的增殖情况，确定M—NFS-60细胞对Alamar Blue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；⑧M—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48h，测定细胞增殖情况。结果：①M—NFS-60细胞的接种密度为5×10^4cells／mL时，对AlamarBlue的还原能力最强，孵育时间8h后，M—NFS-60细胞对Alamar Blue的还原率开始增强，72h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48h，细胞的增殖率（％）分别为121．58±2．62（113）、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）；113、Ⅲ3、Ⅳ分别与IL-3联用处理细胞48h后的增殖率分别为163．98±9．20（113±IL3）、159．89±8．31（Ⅳ-IL3）、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。

The Accelerating-effect on Irradiated M-Nfs-60 Cells Proliferation of Effective Peptide Components from Scorpion Venom

Objective：To observe promoting effects of scorpion venom peptide （SVP） and IL-3 on M-CSF- dependent cell line M-NFS-60 proliferation, as well as its effects on irradiated M-Nfs-60 cells. Methods： By using AlamarBlue viability/proliferation assay to detected cell proliferation situations. （1）Selecting the 10 cells density and detecting of cell proliferation at 0 h, 2 h, 4 h, 5.5 h and 20 h respectively to determine the best cell density and incubation time of M-NFS-60 cells response to AlamarBlue; （2）By using the best cell densityand incubation time to observe the proliferation-accelerating role of 10 SVP components on M-NFS-60 cells, and then to screening the effective component; （3）M-NFS-60 cells were 60Coγ-ray irradiated and then given saline, SVP, IL-3 or SVP ＋ IL-3 treatment for 48 h ours. The rate of cell proliferation was detected. Results： （1）The strongest ability to reduce AlamarBlue wasobserved at the cell density of 5 × 10^4cells/ml. When the incubating time from 8 h to 72 h, the rate of M-NFS-60 cells to reduce the AlamarBlueTM was gradually increased in a linear manner; （2）SVP Ⅱ3 and Ⅳ can significantly promote the M-NFS-60 cell proliferation,which was screened from the 10 SVP component; （3）The irradiated M-NFS-60 cells were treated by SVPⅡ3 ,Ⅲ3 ,Ⅳ ,or IL-3 ,cell proliferating rate （ % ） were 121.58 ±2.62 （Ⅱ3） ,123.39 ±4.45 （ Ⅲ3 ）, 123.51 ± 5. 037 （ Ⅳ）, and 140.12 ±1.68 （ IL-3 ）. Compared with the blank control group （ 100.130 ±. 006 ）, the difference was significant （P 〈 0.01 ）. Combined treatment of SVP and IL-3, the cells proliferating rates were 163. 98 ± 9.20（ Ⅱ3 ± IL3 ）, 159.89 ± 8.31 （ Ⅳ ＋ IL3 ） and 148.92 ± 9.74 （ Ⅲ3 ＋ IL3 ）. Compared with SVP treatment group, the difference was significant （ P 〈 0.05 ）. Conclusions： There were effective peptide components to accelerating M-NFS-60 cells proliferation in scorpion venom. The SVP pepfides could significantly promote M-NFS-60 cells proliferation after it was irradiated and the peptides act synergistically with IL-3.