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目的观察蝎毒增殖多肽(Buthus martensii Karsch proliferation peptide,BmKpp)对X射线辐射后大鼠骨髓间充质干细胞(rat bone-marrow-derived mesenchymal stem cells,rBMSCs)生长的影响。方法利用全骨髓细胞贴壁法分离纯化rBMSCs,体外扩增传代,取第3代至第8代细胞为实验对象。流式细胞术对rBMSCs进行细胞表型鉴定后,将细胞设为非辐射组(IR-组)和辐射组(IR+组,总剂量为8.0 Gy、标称剂量率300 MU/min的X射线照射)。CCK-8法检测BmKpp对各组细胞在不同时间的增殖情况。结果体外培养的第3代或第8代rBMSCs的细胞表型均为:CD29+/CD90+/CD34-/CD45-。在BmKpp对X射线辐射后rBMSCs作用的最低有效浓度筛选的实验中,BmKpp 8μg/ml处理组的吸光度(A值)显著高于辐射对照组(P<0.01);BmKpp(8μg/ml)有促进辐射后的rBMSCs增殖的作用,尤以辐射后第8天、第16天、第18天和第20天效果最为明显,其A值与对照组的相比,P均<0.05;BmKpp(8μg/ml)未能显著促进非辐射rBMSCs的增殖,与对照组相比无统计学差异(P>0.05)。结论 BmKpp能促进辐射后的rBMSCs的增殖,从而发挥其促进辐射后造血功能的恢复作用。
Objective To observe the effect of Buthus martensii Karsch proliferation peptide (BmKpp) on the growth of rat bone marrow-derived mesenchymal stem cells (rBMSCs) after X-ray irradiation. Methods The whole rBMSCs were isolated and purified by whole bone marrow adherent method. The passaged passages were expanded in vitro. The third passage to the eighth passage cells were used as experimental subjects. After the cell phenotypes were identified by flow cytometry, the cells were set as non-irradiated group (IR-group) and irradiated group (total dose of 8.0 Gy, with a nominal dose rate of 300 MU / min) ). CCK-8 method was used to detect the proliferation of BmKpp cells at different time points. Results The cell phenotypes of the third generation or the eighth generation rBMSCs cultured in vitro were: CD29 + / CD90 + / CD34- / CD45-. BmKpp 8μg / ml treatment group absorbance (A value) was significantly higher than the radiation control group (P <0.01); BmKpp (8μg / ml) in the BmKpp X-ray irradiation rBMSCs after the most effective concentration of screening experiments, The effect of radiation on the proliferation of rBMSCs was most obvious on the 8th day, the 16th day, the 18th day and the 20th day after irradiation. The value of A was significantly lower than that of the control group (P <0.05) ml) did not significantly promote the proliferation of non-irradiated rBMSCs, compared with the control group, there was no significant difference (P> 0.05). Conclusion BmKpp can promote the proliferation of irradiated rBMSCs and thus play a role in the recovery of hematopoietic function after radiation.