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Mar Drugs
2023 Apr 26;215:. doi: 10.3390/md21050267.
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Proliferative Effect of Aqueous Extract of Sea Cucumber (Holothuria parva) Body Wall on Human Umbilical Cord Mesenchymal Stromal/Stem Cells.
Rasekh P
,
Kameli A
,
Khoradmehr A
,
Baghban N
,
Mohebbi G
,
Barmak A
,
Nabipour I
,
Azari H
,
Heidari Y
,
Daneshi A
,
Bargahi A
,
Khodabandeh Z
,
Zare S
,
Afshar A
,
Shirazi R
,
Almasi-Turk S
,
Tamadon A
.
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Sea cucumber extracts and their bioactive compounds have the potential for stem cell proliferation induction and for their beneficial therapeutic properties. In this study, human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs) were exposed to an aqueous extract of Holothuria parva body walls. Proliferative molecules were detected using gas chromatography-mass spectrometry (GC-MS) analysis in an aqueous extract of H. parva. The aqueous extract concentrations of 5, 10, 20, 40, and 80 µg/mL and 10 and 20 ng/mL of human epidermal growth factor (EGF) as positive controls were treated on hUC-MSCs. MTT, cell count, viability, and cell cycle assays were performed. Using Western blot analysis, the effects of extracts of H. parva and EGF on cell proliferation markers were detected. Computational modeling was done to detect effective proliferative compounds in the aqueous extract of H. parva. A MTT assay showed that the 10, 20, and 40 µg/mL aqueous extract of H. parva had a proliferative effect on hUC-MSCs. The cell count, which was treated with a 20 µg/mL concentration, increased faster and higher than the control group (p < 0.05). This concentration of the extract did not have a significant effect on hUC-MSCs' viability. The cell cycle assay of hUC-MSCs showed that the percentage of cells in the G2 stage of the extract was biologically higher than the control group. Expression of cyclin D1, cyclin D3, cyclin E, HIF-1α, and TERT was increased compared with the control group. Moreover, expression of p21 and PCNA decreased after treating hUC-MSCs with the extract. However, CDC-2/cdk-1 and ERK1/2 had almost the same expression as the control group. The expression of CDK-4 and CDK-6 decreased after treatment. Between the detected compounds, 1-methyl-4-(1-methyl phenyl)-benzene showed better affinity to CDK-4 and p21 than tetradecanoic acid. The H. parva aqueous extract showed proliferative potential on hUC-MSCs.
Figure 1. Compounds 1–6 isolated from with proliferative biological activity property in sea cucumber (Holothuria parva), including 1,2-benzene dicarboxylic acid, diisooctyl ester (1), 3,4-dihydro-1h-isoquinoline-2-carboxamidine hydrochloride (2), 8-amino-6-methoxyquinoline (3), E-2, 3-epoxycarane (4), cyclooctaneacetic acid, 2-oxo- (5), and isopinocarveol (6).
Figure 2. Characterization of human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs). (A) hUC-MSCs’ morphology. Flow cytometric histograms showing the immunophenotype of hUC-MSCs. (B,C) hUC-MSCs are negative for the hematopoietic line markers CD34 and CD45. (D–F) Analyzed hUC-MSCs are positive for CD44, CD73, and CD90, which are considered to be markers of MSCs. FITC, fluorescein isothiocyanate; PE, phycoerythrin; Cy7, Sulfo-Cyanine7; APC, Allophycocyanin. (G–I) Differentiation of hUC-MSCs to adipogenic differentiation, in which the cells were stained with oil red O, osteogenic differentiation, in which the cells were stained with alizarin red, and chondrogenic differentiation, in which the cells were stained with Alcian blue.
Figure 3. Mean and standard errors of cell proliferation rate MTT (A), cell count (B), and cell viability (C) of human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs) after exposure to the aqueous extract of sea cucumber (Holothuria parva) or epidermal growth factor (EGF). * p < 0.05; ** p < 0.01; *** p < 0.001.
Figure 4. Cell cycle assay of human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs) after exposure to aqueous extract of sea cucumber (Holothuria parva) (A), epidermal growth factor (EGF) (B) or control (C). (D), Proportion of percent of cell number in four stages of cell cycle in three groups.
Figure 5. Expression of proliferation-related proteins after treating human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs) with aqueous extract of sea cucumber (Holothuria parva) or epidermal growth factor (EGF).
Figure 6. Aqueous extract of sea cucumber (Holothuria parva) induces proliferation in human umbilical cord mesenchymal stromal/stem cells (hUC-MSCs). Small colorful dots, red lighting, blue arrows, and red hammer-headlines represent the extract bioactive molecules detected by the docking technique, the extract-induced stimulus, direct reactions, and each inhibiting reaction, respectively.
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