转化生长因子β及重组人骨形成蛋白2对雪旺细胞生物学行为影响的实验研究

STUDY ON BIOLOGICAL EFFECT OF TRANSFORMING GROWTH FACTOR β AND RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN 2 ON THE SCHWANN CELL

目的研究转化生长因子β(transformin ggrowth factorβ,TGF-β)、重组人骨形成蛋白2(recom binant humanbonemor phogenetic protein2,rhBMP-2)对雪旺细胞生物学行为的影响。方法体外培养SD乳鼠雪旺细胞,按5×103/孔接种96孔板,分为3组。A组:加入50ng/mlTGF-β;B组:加入50ng/mlrhBMP-2;C组不作任何处理,作为空白对照组。采用MTT法每天取4孔连测9d细胞数,并绘制生长曲线;流式细胞仪测定细胞周期以观察雪旺细胞增殖情况;ELISA双夹心法测定培养液中神经生长因子(nervegrowth factor,NGF)含量。结果MTT法检测显示A、B组细胞生长曲线远离C组,逐渐上升到第8、9天差异明显,A组较B组上升趋势更明显,差异有统计学意义(P<0.05)。流式细胞仪测定显示A组和B组细胞增殖指数较C组高(P<0.05)。ELISA法检测A组上清NGF含量最高,各组间比较差异有统计学意义(P<0.05)。结论外源性TGF-β、rhBMP-2均能促进雪旺细胞增殖和分泌NGF的功能,而TGF-β更优于rhBMP-2。

Objective To study biological effect of transforming growth factor β（TGF-β） and recombinant human bone morphogenetic protein 2 （rhBMP 2） on the Schwann cell（SC） in vitro, Methods Cultured SC from newborn SD rats were implanted at 5×10^3/well in 96-well-plate （36 wells in each group, altogether 3 groups）：TGF-β group （group A） treated with 50 ng/ml TGF-β rhBMP 2 group （group B） treated with 50 ng/ml rhBMP-2 and control group （group C）. SC proliferation activity was assessed by MTT and flow cytometry （FCM） methods, and nerve growth factor （NGF） synthesis in SC culture media was detected by ELISA method. Results MTT observation indicated that there was significant difference in the growth curve among 3 groups until the 8th and 9th day, Group A had more obvious rising tendency than group B and group C, FCM observation indicated that the proliferation index of group A and group B was higher than that of group C（P〈0, 05）, ELISA observation indicated that there was signiSicant difference in the NGF concentration of the culture medium among the 3 groups（P〈0.05）, Group A had the highest NGF concentration. Conclusion Exogenous TGF-β and rhBMP-2 can promote SC＇s ability to proliferate NGF, but TGF-β is more effective than rhBMP 2.