Objective:To study the intervention effect of highly active mesenchymal stem cells (HA-MSCs) on the structure and function of the ovary in aging macaques.Methods:Macaque HA-MSCs and bone marrow mesenchymal stem cells (BMSCs) were isolated and expanded from young macaque bone marrow using TeSR-E8 medium and DMEM/F12 medium containing 10% fetal bovine serum, respectively. Cell surface markers were detected by flow cytometry, and adipogenic, osteogenic, and chondrogenic differentiation capacities were identified. These two kinds of cells were then infused intravenously into ovarian senescent macaques at a dose of 1×107 cells/kg, once a day for 3 consecutive days. The peripheral blood anti-Mullerian hormone (AMH) levels were measured before and 1, 2, 3 and 5 months after treatment, and the macaques were euthanized at the 5th month after treatment. The histopathological structure of the ovaries was observed by HE staining, and the degree of ovarian fibrosis alteration was evaluated by Masson staining. Immunohistochemical staining was employed to detect the expression levels of ageing-associated markers p16 and p21 in the ovaries.Results:HA-MSCs are short and fusiform with a high nuclear/cytoplasmic ratio and uniform morphology. BMSCs are large in size, showing typical fibroblast morphology, with few a small number of miscellaneous cells. The adipogenic, osteogenic, and chondrogenic differentiation capacities of HA-MSCs are all superior to those of BMSCs. Compared with the aged model group, the HA-MSCs treatment group showed an increase in AMH levels [ (1.85 ± 0.21) vs (0.29 ± 0.15) ng/mL] within two months post-treatment, with ovarian tissues exhibiting follicles of various stages and a decreased degree of fibrosis (all P < 0.001) . Compared to the aged model group, the BMSCs treatment group demonstrated an increase in AMH levels [ (1.26 ± 0.28) vs (0.36 ± 0.16) ng/mL] one month post-treatment. When compared with the HA-MSCs treatment group, the BMSCs treatment group showed a decrease in AMH levels [ (1.85 ± 0.21) vs (0.60 ± 0.20) ng/mL] two months after treatment. The ovarian tissues of this group revealed a small number of immature and primordial follicles, along with a reduced dreduction in fibrosis (all P < 0.001) . The elderly model group had the lowest AMH levels, with occasional atretic follicles and extensive fibrosis in the ovarian tissues. The expression levels of p16 and p21 proteins in the ovaries were highest in the elderly model group, lowest in the HA-MSCs treatment group, and intermediate in the BMSCs treatment group.Conclusion:HA-MSCs can increase the reserve capacity of the aging ovary, and significantly mitigate ovarian fibrosis and the expression levels of ovarian aging-related proteins, and restart the follicle formation function. HA-MSCs transplantation therapy can be a better intervention for the treatment of ovarian aging than BMSCs.
Macrophages are crucial in maintaining homeostasis under cardiac physiological conditions and tissue repair after injury. While the heart is in homeostasis, myocardial tissue resident macrophages promote coronary artery development and electrical conduction within the atrioventricular node, remove aging and dying cells, and conduct immune surveillance. Macrophages recruited after myocardial infarction produce pro-inflammatory and anti-inflammatory mediators such as cytokines, chemokines, matrix metalloproteinases and growth factors that devour dead cells and stromal debris and promote angiogenesis and scarring. This review focuses on the origin and heterogeneity of resident macrophages in myocardial tissue and the different roles of CCR2+ and CCR2? cardiac macrophage subsets under steady-state conditions and after myocardial infarction.
The clinical application of umbilical cord blood as a source of hematopoietic stem cells (HSC) for transplantation is limited by the low dose of CD34+ cells, increased risk of graft failure and slow hematopoietic recovery. Omidubicel is a kind of Umbilical cord blood cell therapy product that expands hematopoietic stem cells in vitro by nicotinamide (NAM) . Compared with unmanipulated Umbilical cord blood, it has faster implantation speed and a lower infection rate in allogeneic hematopoietic stem cell transplantation. Here, we focus on reviewing the relevant data on the clinical application of Omidubicel in malignant hematological diseases, and affirm the early benefits of this amplification strategy. However, its long-term impact on patients still requires large-scale clinical studies. The emergence of Omidubicel pushes expanded umbilical cord blood transplantation to the forefront of transplantation treatment and regenerative medicine.
Objective:To investigate the mechanism of miR-301a-3p in proliferation and apoptosis of human ovarian granulocyte KGN cells.Methods:The expression level of miR-301a-3p was detected by RT-qPCR in tissues and peripheral blood of 38 patients with polycystic ovary syndrome (PCOS) and human ovarian granule KGN cells. KGN cells were divided into control group, anti-miR-NC group and anti-miR-301a-3p group; the expression of miR-301a-3p was detected by RT-qPCR; cell proliferation and apoptosis were detected by MTT and flow cytometry; clone formation ability were detected by plate test; the expressions proliferating cell nuclear antigen (PCNA) , B-cell lymphoma 2 (Bcl2) , BCL2-associated X protein (Bax) , phospho-p38 mitogen-activated protein kinase (p-p38 MAPK) and phospho-extracellular signal-regulated kinase 1/2 (p-ERK1/2) were detected by Western blot. SB203580, an inhibitor of MAPK signaling pathway, was used to investigate the effects on the cell proliferation and apoptosis by the above methods.Results:Compared with the control group, the expression of miR-301a-3p was increased in PCOS tissues (1.08 ± 0.22 vs 2.65 ± 0.44) and peripheral blood (1.05 ± 0.22 vs 2.54 ± 0.47) , and the difference was statistically significant (both P < 0.05) . Compared with IOSE80a cells of human normal ovarian epithelium, the expression of miR-301a-3p in human ovarian granule KGN cells was increased (0.99 ± 0.10 vs 2.38 ± 0.38) , and the difference was statistically significant (P < 0.05) ; down-regulating miR-301a-3p decreased the cell activity, the number of cloned cells, increased the apoptosis rate and the expression of Bax protein, and decreased the expression levels of PCNA, Bcl-2, p-p38 MAPK and p-ERK1/2 proteins and the difference was statistically significant (all P < 0.05) . Compared with the anti-miR-301a-3p group, the cell number and cell activity of the anti-miR-301a-3p+SB203580 clone were significantly decreased, and the protein expression levels of PCNA and Bcl-2 were significantly decreased, the apoptosis rate and the expression level of Bax protein were significantly increased, and the difference was statistically significant (all P < 0.05) .Conclusions:miR-301a-3p is highly expressed in human ovarian granulosa cells, and down-regulation of miR-301a-3p can inhibit the proliferation and induce apoptosis of human ovarian granulosa KGN cells, and its mechanism may be related to inhibition of MAPK signaling pathway.
Objective:To analyze the influence of Celastrol on lipopolysaccharide (LPS) -induced alveolar epithelial cell injury and the role of stromal cell-derived factor-1 (SDF-1) /chemokine receptor 4 (CXCR4) pathway in it.Methods:Human alveolar type Ⅱ epithelial cell line A549 cells were cultured and treated with 10 μg/mL LPS for 24 h. Before LPS treatment, treat them with final concentrations of 25 mmol/L, 100 mmol/L of celastrol or 100 mmol/L of celastrol and 100 μg/L of AMD3100 (CXCR4 inhibitor) for 2 hours. The viability of A549 cells was determined by the CCK-8 method, apoptosis of A549 cells was measured by the TUNEL method, the LDH of A549 cells was determined by kits, the levels of tumor necrosis factor (TNF) -α, interleukin-6 (IL-6) and IL-10 in A549 cells were detected by ELISA, and the expression of SDF-1 and CXCR4 protein were as detected by Western blot. One-way analysis of variance was used for statistical comparison among multiple groups, and the SNK-q method was used for further pairwise statistical comparison.Results:Compared with the control group, the positive rate of TUNEL [ (65.78 ± 8.21) % vs (4.23 ± 0.77) %], the apoptosis rate [ (21.38 ± 2.70) %vs (7.64 ± 0.95) %], cleaved caspase-3, Bax protein, LDH, TNF-α, IL-6, SDF-1 (2.02 ± 0.25 vs 0.75 ± 0.10) , and CXCR4 (1.59 ± 0.19 vs 0.71 ± 0.08) protein were increased in A549 cells in LPS group (P < 0.05) , and the survival rate, Bcl-2 protein and IL-10 expression decreased (P < 0.05) ; compared with the LPS group, the positive rate of TUNEL [ (52.51 ± 6.54) %, (41.37 ± 5.17) %vs (65.78 ± 8.21) %], the apoptosis rate was [ (17.62 ± 2.45) %, (12.37 ± 1.54) %vs (21.38 ± 2.70) %], cleaved caspase-3, Bax protein, LDH, TNF-α, IL-6, SDF-1 (1.69 ± 0.20, 1.32 ± 0.18 vs 2.02 ± 0.25) , CXCR4 (1.31 ± 0.16, 1.05 ± 0.13 vs 1.59 ± 0.19) protein in A549 cells in the low-dose Celastrol group and high-dose Celastrol group significantly reduced, and the survival rate, Bcl-2 protein and IL-10 expression significantly increased (P < 0.05) ; compared with the high-dose Celastrol group, the positive rate of TUNEL[ (24.28 ± 4.28) %vs (41.37 ± 5.17) %], the apoptosis rate [ (9.91 ± 1.25) %vs (12.37 ± 1.54) %], and cleaved caspase-3, Bax protein, LDH, TNF-α, IL-6, SDF-1 (0.95 ± 0.13 vs 1.32 ± 0.18) , and CXCR4 (0.78 ± 0.09 vs 1.05 ± 0.13) protein of A549 cells in the high-dose Celastrol + AMD3100 group decreased, and the survival rate, Bcl-2 protein and IL-10 expression significantly increased (P < 0.05) .Conclusion:Celastrol can reduce LPS-induced apoptosis and inflammation of A549 cells and improve cell survival, which may be related to the inhibition of the SDF-1/CXCR4 pathway.
