RESUMO
Objective:To explore the mechanism of Bushen Huayu Shengxin decoction in delaying senescence of bone mesenchymal stem cells(BMSCs) by improving cellular microenvironment and regulating p16/pRb and p53/p21 signaling pathways. Method:The cells were cultured in serum-free 1640 medium and hypoxic cell workstation for 24 hours to establish the cell model of ischemic-hypoxic microenvironment in vitro, then randomized into control group (with complex medium), model group (with complete medium), and treatment group (with serum medium-containing Bushen Huayu Shengxin decoction), and all were cultured in hypoxic cell workstation for 24 hours. The normal group was added with control culture for complete medium, The cell cycle of BMSCs was detected by flow cytometry, the expressions of p16INK4a, p53, p21 and Survivn, cysteine aspartic acid protease-3 (Caspase-3), polyadenosine diphosphate ribose polymerase (PARP) mRNA were analyzed by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the levels of β-catenin protein and glycogen synthase kinase-3β（GSK-3β） protein were detected by Western blot. Result:Compared with the normal group, the proportion of S phase cells increased, while that at the G0/G1 phase decreased significantly in the model group (P<0.05). Compared with the model group, the proportion of S phase decreased, whereas that at the G0/G1 phase gradually increased in the treatment group (P<0.05). Compared with the normal group, mRNA expressions of p16INK4a, p53, p21 and Survivn, Caspase-3, PARP in the model group increased significantly (P<0.05). Compared with the model group, mRNA expressions of p16INK4a, p53, p21 and Survivn, Caspase-3, PARP in the treatment group decreased significantly (P<0.05). Compared with the normal group, protein expressions of β-catenin and GSK-3β in the model group increased significantly (P<0.05). Compared with the model group, protein expressions of β-catenin and GSK-3β in the treatment group decreased significantly (P<0.05). Conclusion:Bushen Huayu Shengxin decoction could delay the senescence of BMSCs by improving ischemic-hypoxic microenvironment and regulating p16/pRb and p53/p21 signaling pathways.
RESUMO
<p><b>OBJECTIVE</b>To investigate the mechanism that the formulas for activating blood and resolving stasis can regulate hemopoietic stem cell to produce new blood.</p><p><b>METHOD</b>Rats were established animal model of acute cerebral infarction by referencing Olivette' method. They were randomly divided into model group, the group of the high, middle, low dose of the formulas for activating blood and resolving stasis. Each group and then wasrandomly divided into subgroups by 1, 3, 7, 14, 28 d. Xuesaitong capsule was formulated into 20, 40, 60 g x L(-1) with normal saline. The rats were given gavage drugs once a day until the experient ended, and the model group was administrated by intragastrical perfusion of normal saline. ELISA was used to detect the expression of SCF in peripheral blood and bone marrow among different groups at different time points. Flow cytometry was used to observe the changes of CD117 in blood and bone marrow.</p><p><b>RESULT</b>The CD117+ HSC and SCF concentration in peripheral blood and bone marrow of model group were increasing during 1-14 d,there was a peak on the 14th day, then the expression was reducing. CD117+ HSC and SCF concentration rising trend in the group of the high, middle dose of the formulas for activating blood and resolving stasis was preceded model group (P < 0.05).</p><p><b>CONCLUSION</b>Activating blood and resolving stasis can regulate hemopoietic stem cell to produce new blood, and it is through the regulation of CD117+ HSC number to achieve the purpose.</p>