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Inhibitory effect of total saponins of Panax notoginseng on rat bone marrow mesenchymal stem cell apoptosis

OU Yong-Fang, FU Xue-Kun, MEI Xing-Sha, ZHENG Hui-Zhen^*

1Department of Physiology, College of Basic Medicine, Guangdong Medical University, Dongguan 523808, China； 2Pathological Diagnosis and Research Center, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China

Abstract

The study was aimed to investigate the effects of total saponins of Panax notoginseng (tPNS) on cobalt chloride (CoCl2)-induced apoptosis of rat bone marrow mesenchymal stem cells (rBMSCs) and the underlying mechanism. rBMSCs were isolated by density gradient centrifugation from Sprague Dawley (SD) rats. After being incubated with different concentrations of tPNS (1, 10, 100 μg/mL) for 48 h, the rBMSCs were stained with EdU and PI for proliferation and cell cycle assay, respectively. CoCl2 group was
treated with 300 μmol CoCl2 for 24 h, and different concentrations tPNS groups were treated with 300 μmol CoCl2 plus 1, 10 or 100 μg/mL tPNS. After Annexin V-FITC/PI staining, flow cytometry was applied to measure the cell apoptosis. For mitochondrial
membrane potential assay, rhodamine123 and Hoechst33342 staining were used. qRT-PCR was applied to analyze gene expression of Bcl-2 family. The results showed that the proliferation rates of the three concentrations tPNS groups were all higher than that of the control group (all P < 0.05). Compared with control group, only 100 μg/mL tPNS group exhibited increased cell percentage of S and G2 phase. Compared with that in control group (without CoCl2), the apoptotic rate was increased by 14.2% in CoCl2 group. And the apoptotic rates were reduced by 14.4%, 12.8% and 13.9% in three concentrations tPNS groups, compared with that in CoCl2 group (all P < 0.01). CoCl2 could decrease the mitochondrial membrane potential, while different concentrations of tPNS reversed the inhibitory effect of CoCl2. Bcl-2 and Bcl-xl mRNA expressions in all tPNS groups were higher than those in CoCl2 group (all P < 0.05). Moreover, 10 and 100 μg/mL tPNS groups showed lower ratios of Bax/Bcl-2, compared with CoCl2 group. The results suggest that tPNS protects the rBMSCs against CoCl2-induced apoptosis through improving the cell mitochondrial membrane potential, up-regulating the expressions of anti-apoptosis genes Bcl-2 and Bcl-xl, and reducing the Bax/Bcl-2 gene expression ratio.

Key words: total saponins of Panax notoginseng (tPNS); bone marrow mesenchymal stem cells; cell proliferation; cell cycle; apoptosis; mitochondrial membrane potential; rat

Received: 　　Accepted:

Corresponding author: 郑惠珍　　E-mail: zhenghz6@gdmc.edu.cn

Citing This Article：

OU Yong-Fang, FU Xue-Kun, MEI Xing-Sha, ZHENG Hui-Zhen. Inhibitory effect of total saponins of Panax notoginseng on rat bone marrow mesenchymal stem cell apoptosis. Acta Physiol Sin 2016; 68 (3): 285-292 (in Chinese with English abstract).