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目的探讨人参皂苷Rg1(ginsenoside Rg1)对缺氧复氧BMSCs增殖和凋亡的影响,并探讨其可能机制。方法实验分为BMSCs正常对照组、BMSCs缺氧复氧组(Model组)、人参皂苷Rg11×10~(-7)、1×10~(-6)、1×10~(-5)mol/L处理组、尼莫地平2.5×10~(-7)mol/L处理组(阳性对照组)。各组分别于缺氧复氧12h前加入完全培养基(正常对照组、Model组)、人参皂苷Rg1(人参皂苷Rg1各处理组)、尼莫地平(阳性对照组)。建立缺氧复氧BMSCs模型。采用TUNEL法检测各组的细胞凋亡率,免疫荧光和免疫印迹技术定性定量检测各组增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、Bcl-2、Bax的表达情况。结果TUNEL结果显示,BMSCs正常对照组未见明显凋亡细胞,Model组可见明显的凋亡细胞,与Model组比较,其余各处理组凋亡细胞数量均减少,以人参皂苷Rg1(1×10~(-5)mol/L)组最为明显。免疫荧光和免疫印迹结果显示,BMSCs正常对照组可见少量的PCNA、bcl-2、bax表达;Model组的PCNA、bcl-2的表达减少,但bax的表达显著增高,bcl-2/bax比值降低;与Model组比较,人参皂苷Rg1各组PCNA、bcl-2表达均呈不同程度的增高,而bax表达呈现相反的趋势,bcl-2/bax比值增高,以Rg1(1×10~(-5)mol/L)组最为明显。结论人参皂苷Rg1预处理对缺氧复氧BMSCs具有保护作用,其机制可能与下调bax的表达,上调PCNA、bcl-2的表达,抑制BMSCs凋亡和促进BMSCs增殖有关。
Objective To investigate the effect of ginsenoside Rg1 on proliferation and apoptosis of hypoxia-reoxygenation-induced BMSCs and to explore its possible mechanism. Methods The experiment was divided into three groups: BMSCs normal control group, BMSCs hypoxia-reoxygenation group (Model group), ginsenoside Rg11 × 10 -7, 1 × 10 -6, 1 × 10 -5 mol / L treatment group and nimodipine 2.5 × 10 -7 mol / L treatment group (positive control group). Each group was added with complete medium (normal control group, model group), ginsenoside Rg1 (ginsenoside Rg1 treatment group) and nimodipine (positive control group) 12 hours before anoxia and reoxygenation respectively. Establishment of hypoxia-reoxygenation BMSCs model. The apoptotic rate was detected by TUNEL method. The expression of proliferating cell nuclear antigen (PCNA), Bcl-2 and Bax in each group was detected by immunofluorescence and immunoblotting. Results The results of TUNEL showed that there were no obvious apoptotic cells in normal control group and obvious apoptotic cells in Model group. Compared with Model group, the number of apoptotic cells in all the other groups decreased. Ginsenoside Rg1 (1 × 10 ~ (-5) mol / L) group was the most obvious. Immunofluorescence and Western blotting results showed that a small amount of PCNA, bcl-2 and bax expression were observed in normal control group; the expression of bcl-2 and PCNA in model group was decreased, while the bcl-2 / bax ratio was decreased Compared with Model group, the expressions of PCNA and bcl-2 in ginsenoside Rg1 groups all increased to some extent, while the expression of bax showed the opposite trend and the ratio of bcl-2 / bax increased. The expression of Rg1 (1 × 10 -5 ) mol / L) group was the most obvious. Conclusion Ginsenoside Rg1 preconditioning can protect BMSCs against hypoxia-reoxygenation. The mechanism may be related to down-regulating the expression of bax, up-regulating the expression of PCNA and bcl-2, inhibiting the apoptosis of BMSCs and promoting the proliferation of BMSCs.