The effect of lovastatin on cell proliferation and neurotrophic factor expression of bone marrow mesenchymal stem cells in vitro

bageri A, MT Ghorbanian, A. Kosha
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Abstract

Mesenchymal stem cells, Berberis, integrrima, Osteoblast, Differentiatio n. Aim: In recent years using of Bone marrow mesenchymal stem cells (BMSC) in regenerative medicine, tissue engineering and gene therapy is highly regarded. Convenient access, ability to expand and MSC differentiation capacity along with the ability of adhesion to plastic surfaces and in-vitro growth and development are considered as the characteristic feature of these cells. Bone marrow mesenchymal stem cells possess the ability to differentiate into mesodermal lineage, among other adult cells, can be used in tissue engineering and are good candidates for transplantation. Lovastatin as a lowering cholesterol agent and reducing inflammation, as well as antioxidant and, in particular, neuroprotective effects can be effective in the treatment of neurogenic diseases. The aim of this study was to evaluate the effect of lovastatin on survival, proliferation and expression of GDNF and oct4 genes of Bone marrow mesenchymal stem cells. Lovastatin is presumed to exert their neuroprotective effects by inducing neurotrophic factor gene expression and cell proliferation. Material and methods: In this experimental study, we used 4-6 week adult Wistar rats. The BMSCs were isolated from rat femurs and tibias and cultured in α-MEM. The cell pellet was resuspended in α-MEM supplemented with 10% fetal bovine serum (FBS), 1% penicillin and streptomycin and cultured in 25-cm2 culture flasks at a density of 2 × 104 cells and incubated at 37°C and 5% CO2. For lovastatin treatment, we exposed MSCs to 1 μM, 5 μm, 10 μm and 15 μm of lovastatin for 24 h. The survival rate of cells was measured by MTT assay. The growth rate and proliferation of cells at 24 hours after culture were assessed by staining with DAPI. The expression of Oct4 and GDNF factors was also evaluated by RT-PCR. Results: MSCs were attached to culture plate and were quickly proliferated. In the culture plate, these cells were usually appeared in three forms: small spherical, fusiform and fibroblast-like and Piezoelectric Energy Harvesting from Functionally Graded Beams Using Modified Shear Deformation Theories 2 Mechanics of Advanced and Smart Materials Journal 1(2) (2022) 107 – 120 flattened. The results of this study indicate that the proliferation rate at 5, 10 and 15 μM lovastatin showed a significant increase compared to control groups (P<0.5). Gene expression density of gelial derived neurotrophic factors (GDNF) and oct4 genes showed that, there were significant differences between MSCs treatment groups and control group (P<0.5). Cell viability and proliferation rate indicate that experimental groups has a higher proliferation rate than control group. Moreover, results showed an increase in mRNA expression for GDNF and Oct4 compared to the control group (P <0.05). Conclusion:Therefore, lovastatin can be used to improve the culture of mesenchymal stem cells, which is used for transplantation and cell therapy. BMSCs may be a useful therapeutic agent for the treatment of neurodegenerative disorders. هرود ،تفاب و لولس 13 هرامش ، 2 ، لاس 1401 تاحفص ، 107 ات 120
洛伐他汀对体外培养骨髓间充质干细胞细胞增殖及神经营养因子表达的影响
摘要目的:近年来,骨髓间充质干细胞(BMSC)在再生医学、组织工程和基因治疗中的应用受到高度重视。便捷的获取、扩展能力、间充质干细胞分化能力以及与塑料表面的粘附能力和体外生长发育能力被认为是这些细胞的特征。骨髓间充质干细胞具有向中胚层分化的能力,在其他成体细胞中,可用于组织工程,是移植的良好候选者。洛伐他汀作为一种降胆固醇剂和消炎剂,以及抗氧化剂,特别是神经保护作用,可以有效地治疗神经源性疾病。本研究旨在评价洛伐他汀对骨髓间充质干细胞存活、增殖及GDNF和oct4基因表达的影响。洛伐他汀可能通过诱导神经营养因子基因表达和细胞增殖来发挥其神经保护作用。材料与方法:本实验选用4-6周龄成年Wistar大鼠。从大鼠股骨和胫骨分离骨髓间充质干细胞,并在α-MEM中培养。将细胞颗粒重悬于添加10%胎牛血清(FBS)、1%青霉素和链霉素的α-MEM中,以2 × 104个细胞的密度在25 cm2的培养瓶中培养,37℃、5% CO2孵育。对于洛伐他汀,我们将MSCs分别暴露于1 μM、5 μM、10 μM和15 μM的洛伐他汀中24 h,采用MTT法测定细胞存活率。DAPI染色观察培养后24小时细胞的生长速率和增殖情况。RT-PCR检测Oct4和GDNF因子的表达。结果:骨髓间充质干细胞贴壁,增殖迅速。在培养板中,这些细胞通常以三种形式出现:小球形,梭形和成纤维细胞样。利用改进的剪切变形理论从功能梯度梁中收集压电能量2先进与智能材料力学学报1(2)(2022)107 - 120扁平。本研究结果显示,与对照组相比,5、10和15 μM洛伐他汀组细胞增殖率显著升高(P<0.5)。胶质源性神经营养因子(GDNF)和oct4基因表达密度显示,MSCs处理组与对照组差异有统计学意义(P<0.5)。细胞活力和增殖率显示实验组的增殖率高于对照组。与对照组相比,GDNF和Oct4 mRNA表达增加(P <0.05)。结论:洛伐他汀可改善间充质干细胞的培养,用于移植和细胞治疗。骨髓间充质干细胞可能是治疗神经退行性疾病的有效药物。هرود،تفابولولس13هرامش،2،لاس1401تاحفص،107ات120
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