人参皂苷Rg1对白消胺诱导细胞衰老的延缓作用

Ginsenoside Rg1 attenuates Busulfan-induced premature senescence

目的探讨人参皂苷Rg1(Ginsenoside Rg1,Rg1)对白消胺(Busulfan,BU)诱导的人胚肺成纤维细胞衰老的延缓作用。方法将人胚肺成纤维细胞(<24代)随机分为空白对照组(常规培养)、BU组(以120μmol/L BU处理复制细胞衰老模型)、BU+Rg1组(120μmol/L BU+10μmol/L Rg1)、Rg1预+BU组(10μmol/L Rg1预处理后加入120μmol/L BU)和Rg1预+BU+Rg1组(10μmol/L Rg1预处理后加入120μmol/L BU,再加入10μmol/L Rg1),通过倒置显微镜观察细胞形态的变化,细胞增殖试验检测细胞增殖情况,流式细胞术分析细胞周期的分布;半乳糖苷酶(SA-β-gal)染色法观察细胞衰老情况。结果 BU组细胞胞体扁平、宽大、不规则,出现空泡,而Rg1处理各组细胞的衰老表型出现一定程度的改善,以Rg1预+BU+Rg1组效果最佳,但未完全恢复;BU组细胞增殖能力较空白对照组明显降低,Rg1处理组细胞增殖能力有所恢复,且随着时间的延长效果越明显,其中以Rg1预+BU+Rg1组最佳;Rg1预+BU组和Rg1预+BU+Rg1组G2/M期比例较BU组显著降低(P<0.05),其中以Rg1预+BU+Rg1组降低最明显;Rg1预+BU组和Rg1预+BU+Rg1组衰老细胞数较BU组显著降低(P<0.05)。结论 Rg1能有效对抗BU诱导的细胞早衰,其具体作用机制有待进一步深入研究。

Objective To investigate the role of ginsenoside Rgl in attenuating Busulfan（BU）-induced premature senescence of human embryonic lung fibroblasts. Methods Human embryonic lung fibroblasts within 24 passages were divided into five groups. The fibroblasts in blank control group were subjected to routine culture, while those in BU group were treated with 120 μmol/L BU to copy cell model of premature senescence, those in BU ＋ Rgl group with 120 μmol/L BU ＋ 10 pμmol/L Rgl, those in Rgl pret reatment ＋ BU group were pretreated with 10 μmol/L Rgl then treated with 120 μmol/L BU, and those in Rgl pretreatment ＋ BU ＋ Rgl group were pretreated with 10 μmol/L Rgl then treated with 120 μmol/L BU and 10 μmol/L Rgl. The fibroblasts in various groups were observed for morphological changes under inverted microscope, determined for proliferation by cell proliferation test, ana- lyzed for cell cycle distribution by flow cytometry, then observed for senescence by SA-β-gal staining. Results The fibroblasts in BU group were flat, wide and in irregular shape, in which vacuoles were observed. However, the premature senescence of fibroblasts treated with Rgl was improved, especially those in Rgl pretreatment ＋ BU ＋ Rgl group, but not recovered completely. The prolifer- ation ability of fibroblasts was significantly lower in BU group than in blank control group, while recovered after treatment with Rgl, in a time-dependent mode, especially in Rgl pretreatment ＋ BU ＋ Rgl group. The percentages of fibroblasts at G2/M phase in Rgl pretreatment ＋ BU and Rgl pretreatment ＋ BU ＋ Rgl groups, especially those in the latter, were significantly lower than that in BU group （P 〈 0. 05）. The counts of senescent fibroblasts were significantly lower in Rgl pretreatment ＋ BU and Rgl pretreatment ＋ BU ＋ Rgl groups than in BU group （P 〈 0, 05）. Conclusion Rgl attenuated the premature senescence of human embryonic lung fibroblasts induced by BU, of which mechanism was to be further studied.