HUANG Yi-zhi, LIU Tai, HUANG Ming-zheng, et al. Effect of Shuxue Tongmai Capsule-containing Serum on Apoptosis of Rat Bone Marrow Mesenchymal Stem Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(3): 154-159.
DOI:
HUANG Yi-zhi, LIU Tai, HUANG Ming-zheng, et al. Effect of Shuxue Tongmai Capsule-containing Serum on Apoptosis of Rat Bone Marrow Mesenchymal Stem Cells[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(3): 154-159. DOI: 10.13422/j.cnki.syfjx.2018030154.
Effect of Shuxue Tongmai Capsule-containing Serum on Apoptosis of Rat Bone Marrow Mesenchymal Stem Cells
Objective:To investigate the effect and mechanism of Shuxue Tongmai capsule-containing serum on apoptosis in rat bone marrow mesenchymal stem cells (BMSCs). Method:BMSCs were cultured with different concentrations of H2O2 in vitro to induce apoptosis
and then 1.0 mmol·L-1 optimum concentration was used for the experiment. First
Shuxue Tongmai capsule-containing serum were prepared at low (1.81 g·kg-1)
middle (3.62 g·kg-1) and high (7.24 g·kg-1) doses. Then the third-generation BMSCs were divided into 5 groups in random. The first group was control group
which only added with normal rat serum with a volume fraction of 15% in culture medium. The second group was model group
which was added with normal rat serum with a volume fraction of 15% in culture medium and the apoptosis cells were induced with H2O2.The other three groups were drug-contained serum groups
which were respectively prepared at low (0.32 g·kg-1)
middle (0.64 g·kg-1) and high (1.28 g·kg-1) doses
and added with drug-contained serum with a volume fraction of 15%
and the apoptosis cells were induced with H2O2 in each group. After 24 hours
the cells' relative survival rate was detected by 3-(4
5-dimethyl-2-thiazolyl) -2
5-diphenyl-2-H-tetrazolium bromide (MTT) method
and the apoptosis rate was evaluated by Annexin V-FITC/PI flow cytometry method. And cysteine aspartate protease-3 (Caspase-3) and B-cell lymphoma-2 (Bcl-2) mRNA expressions were assayed by Real-time PCR (Real-time PCR). Result:H2O2 could imitate the ischemic anoxic environment in vitro and induce the apoptosis of BMSCs. The MTT detection results showed that the cells' survival rates of Shuxue Tongmai capsule-containing serum groups prepared at low
medium and high doses were significantly higher than that in the model group (P<0.01)
and the survival rate increased with the dose concentration (P<0.05). And the AnnexinV-FITC/PI flow cytometry results showed that the drug-containing serum of Shuxue Tongmai capsule could decrease the apoptosis rates of BMSCs induced with H2O2 (P<0.01). The Real-time PCR results showed that the induction with H2O2 could increase BMSCs' Caspase-3 mRNA expression and reduce their Bcl-2 mRNA expression. The treatment of Shuxue Tongmai capsule-containing serum could decrease Caspase-3 mRNA expression (P<0.01)
and increase Bcl-2 mRNA expression (P<0.01). Conclusion:Shuxue Tongmai capsule-containing serum can negatively regulate Caspase-3 gene by increasing Bcl-2 gene expression
so as to inhibit the apoptosis of BMSCs caused with H2O2.