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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(P<0.01)。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(P<0.01)。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。
Objective: To observe the protective effect of the femoral head necrosis capsule-containing serum on the apoptosis of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods: BMSCs from rats were isolated, screened and cultured in vitro to prepare the bone marrow-derived osteonecrosis of the femoral head treated with 0.36, 0.72 and 1.44 g · kg-1. The rats in the model group (20% normal rat serum culture medium and whole body) were added into the blank group (adding the culture medium without rat serum), the normal group (adding the 20% normal rat serum culture solution) Trans retinoic acid induced apoptosis) and traditional Chinese medicine serum low, medium and high dose groups (during the culture were added to the corresponding dose, the volume fraction of 20% of femoral head necrosis capsule containing serum-containing serum and all-trans retinoic acid Induced apoptosis), a total of 6 groups, the relative survival rate of each group was detected by MTT method, the cell cycle of each group was detected by flow cytometry, and compared. Results: Compared with normal group, the relative survival rate of model group and traditional Chinese medicine serum in low, medium and high dose groups decreased, the proportion of G1 phase increased and the proportion of G2 and S phase decreased, the difference was significant (P <0.01). Compared with the model group, the relative survival rate of Chinese medicine low, medium and high dose groups increased, the proportion of G1 phase decreased, the proportion of G2 and S phase increased, and with the increase of Chinese medicine dosage, the difference was significant (P < 0.01). CONCLUSION: Capsule-containing serum from femoral head necrosis can increase the survival rate of BMSCs in rats and protect against apoptosis induced by all-trans retinoic acid.