股骨头坏死愈胶囊含药血清对大鼠骨髓间充质干细胞凋亡的保护作用

目的:观察股骨头坏死愈胶囊含药血清对体外大鼠骨髓间充质干细胞(BMSCs)凋亡的保护作用。方法:在体外分离、筛选、培养大鼠BMSCs细胞,制备0.36,0.72,1.44 g·kg-1灌胃处理得到的股骨头坏死愈胶囊含药血清作用后,选择第3代大鼠BMSCs,实验分为空白组(加入不含大鼠血清的培养液),正常组(加入体积分数为20%正常大鼠血清培养液),模型组(加入体积分数为20%正常大鼠血清培养液和全反式维甲酸诱导细胞凋亡)和中药血清低、中、高剂量组(培养过程中分别加入对应剂量、体积分数为20%的股骨头坏死愈胶囊含药血清培养液和全反式维甲酸诱导细胞凋亡),共6组,MTT法检测各组细胞相对存活率,流式细胞仪检测各组细胞周期,进行比较。结果:与正常组比较,模型组,中药血清低、中、高剂量组相对存活率下降,细胞周期中G1期比例上升,G2及S期比例下降,差异有显著性意义(P0.01)。与模型组比较,中药血清低、中、高剂量组相对存活率上升,G1期比例减少,G2及S期比例增加,且随着中药剂量增加而差值明显,差异有显著性意义(P0.01)。结论:股骨头坏死愈胶囊含药血清可提高大鼠BMSCs的存活率,对全反式维甲酸所致的细胞凋亡有保护作用。

Protective Effect of Gugutou Huaisiyu Capsule Containing Serum on Apoptosis in Rat Bone Marrow Mesenchymal Stem Cells

Objective: To observe protective effect of Gugutou Huaisiyu capsule containing serum on apoptosis in rat bone marrow mesenchymal stem cells(BMSCs). Method: The BMSCs were separated, filtrated, cultured, and high, medium and low dose Gugutou Huaisiyu capsule containing serum had harvested from mice under 0.36, 0.72, 1.44 g·kg^-1 Gugutou Huaisiyu capsule gavaged, the third generation BMSCs were chose and divided into blank group (no rat serum in culture medium), normal group (with volume fraction of 20% normal rat serum in culture medium), induction apoptosis group (with volume fraction of 20% normal rat serum and all-trans retinoic acid in culture medium) and traditional Chinese medicine serum low, medium and high dose group (with volume fraction of 20% Gugutou Huaisiyu capsule containing serum of low, medium and high dose respectively, and all-trans retinoic acid in culture medium), a total of six groups.Then cell relative survival rate and cell cycle were detected by MTT method and flow cytometry, respectively. Result: Compared with normal group, the cell relative survival rate of induction apoptosis group, traditional Chinese medicine containing serum low, medium and high dose groups was increased, and G₁ phase increased, G₂ and S phase proportion, the differences were statistically significant (P<0.01). Compared to the induction apoptosis group, the cell relative survival rate in traditional Chinese medicine containing serum low, medium and high dose groups was declined, while the G₁ phase ratio decreases, G₂ and S phase proportion increase, and the difference increased obviously as the dosage increased, the differences were statistically significant (P<0.01). Conclusion: Gugutou Huaisiyu capsule containing serum could improve the survival rate of rat BMSCs, and have protective effect on apoptosis induced by all-trans retinoic acid.