Objective To investigate the mechanisms of ginsenoside Rh2 inducing dendrite formation in melanoma cells. Methods B16 cells were cultured with Rh2, and cells with induced dendrites were counted at the indicated time points. B16 cells were pretreated with methyl-β-cyclodextrin(MβCD)and dendrite induction by Rh2 was examined again. When Rh2 was added,lipid fluidity was measured by fluorescence polarization apparatus using two fluorescent probes, 1, 6-diphenyl-l. 3,5-hexatriene(DPH)and l-[4-(trimethylammonio)phenyl]-6-phenyl-l ,3, 5-hexatriene(TMA-DPH). Results In B16 melanoma cells,Rh2( 18. 5 jumol/L) induced dendrite formation within 2 h, which could be suppressed by depletion of cholesterol by pretreatment with 10 mmol/L M|3CD. Within 2 min of addition,Rh2 changed the fluorescence polarization of DPH,but not of TMA-DPH. Conclusion Rh2 affects the physical properties of cholesterol-regulated membrane lipid bilayers and leads to changes in cellular functions.%目的 探讨人参皂甙Rh2诱导黑色素瘤细胞树突生成的机制.方法 在培养的B16黑色素瘤细胞中加入Rh2,记录其诱导细胞树突生成的时间及细胞数.加入甲基-β-环糊精预处理清除胆固醇后,再次观察对Rh2诱导细胞树突生成的影响.用荧光偏振测量仪检测Rh2对反映膜流动性的DPH(1,6-二苯基1,3,5-已三烯)及TMA-DPH(1-[4-(三甲铵)苯基]-6-苯基1,3,5-已三烯)两种荧光探针的影响.结果 在B16黑色素瘤细胞中,Rh2(18.5 μmol/L)在2h内诱导了树突的生成,而用10 mmol/L的甲基-β-环糊精预处理清除胆固醇后即可抑制Rh2的这种作用.加入Rh2后2min内,Rh2即改变了DPH的荧光偏振,但对TMA- DPH却没有影响.结论 Rh2影响了胆固醇调节的膜脂质双层的生理特性,从而可能进一步导致细胞功能的改变.
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