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目的研究人参皂苷Rg1(ginsenoside Rg1)对低氧-无血清培养诱导的大鼠骨髓间充质干细胞(rBMSC)凋亡的保护作用及机制。方法 rBMSC在1%O2及无血清培养基条件下培养24h,培养液中加或不加入人参皂苷Rg1,分析Rg1的作用。Annexin V-FITC/PI染色检测细胞凋亡率、罗丹明123染色检测线粒体膜电位、比色法检测Caspase-3酶活性,实时荧光定量PCR检测Bcl-2、Bcl-xl、Bad、Bax基因的表达水平。结果与正常培养组(C1组)比较,低氧-无血清培养基组(C2组)细胞凋亡率是C1组的3.24倍,P<0.01;线粒体膜电位明显下降;Caspase-3活性高5.17倍,P<0.01;促凋亡基因Bad和Bax的mRNA表达分别增加1.16倍和0.5倍,P<0.01和P<0.05。人参皂苷Rg110μg/L(Rg1-L组)、100μg/L(Rg1-M组)和500μg/L(Rg1-H组)细胞凋亡率分别较C2组下降19.4%、35.1%和65.5%,均P<0.05。Caspase-3活性在Rg1-L组与C2组之间无明显差异,但Rg1-M组和H组较C2组分别降低20%和21%,均P<0.05。BadmRNA表达,Rg1-L组、Rg1-M组和Rg1-H组较C2组分别减少36.4%、50.3%和45.5%,P<0.05或P<0.01;BaxmRNA表达分别减少10.5%、56.7%和50.1%,P<0.05或P<0.01。Bcl-2mRNA表达,Rg1-M组和Rg1-H组较C2组分别增加53.4%和66.1%,均P<0.05。Bcl-xlmRNA表达水平组间比较差异无统计学意义。结论人参皂苷对低氧-无血清培养诱导的rBMSC凋亡有保护作用。
Objective To study the protective effect and mechanism of ginsenoside Rg1 on apoptosis of rat bone marrow mesenchymal stem cells (rBMSCs) induced by hypoxia-serum-free culture. Methods rBMSCs were cultured in 1% O2 and serum-free medium for 24 h. The effect of Rg1 was analyzed by adding or not adding ginsenoside Rg1 to the culture medium. Mitochondrial membrane potential was detected by Annexin V-FITC / PI staining, Caspase-3 activity was detected by colorimetric assay, Bcl-2, Bcl-xl, Bad and Bax genes were detected by real- The expression level. Results Compared with normal group (group C1), the apoptotic rate in group C2 was 3.24 times higher than that in group C1 (P <0.01), the mitochondrial membrane potential was significantly decreased, and the activity of Caspase-3 was 5.17 Fold, P <0.01. The mRNA expressions of the pro-apoptotic genes Bad and Bax increased by 1.16-fold and 0.5-fold, respectively, P <0.01 and P <0.05. The apoptotic rates of ginsenoside Rg110μg / L (Rg1-L group), 100μg / L (Rg1-M group) and 500μg / L (Rg1-H group) were decreased by 19.4%, 35.1% and 65.5% P <0.05. Caspase-3 activity had no significant difference between Rg1-L group and C2 group, but Rg1-M group and H group were 20% and 21% lower than C2 group respectively (P <0.05). BadmRNA expression was decreased by 36.4%, 50.3% and 45.5% in Rg1-L group, Rg1-M group and Rg1-H group compared with C2 group, P <0.05 or P <0.01; Bax mRNA expression decreased by 10.5%, 56.7% and 50.1 %, P <0.05 or P <0.01. Bcl-2mRNA expression increased by 53.4% and 66.1% in Rg1-M group and Rg1-H group compared with C2 group, both P <0.05. Bcl-xlmRNA expression levels between the groups showed no significant difference. Conclusion Ginsenosides have a protective effect on rBMSC apoptosis induced by hypoxia-serum-free culture.