Universitas Syiah Kuala, Faculty of Mathematics and Natural Sciences, Department of Biology, Banda Aceh, Indonesia.
Research Center for Genetic Engineering National Research and Innovation Agency, Cibinong, Indonesia.
Braz J Biol. 2023 May 29;83:e270335. doi: 10.1590/1519-6984.270335. eCollection 2023.
Degenerative diseases, such as osteoporosis, could be treated by stem cells. The aim of this study was to identify the gene expression of bone marrow mesenchymal stem cells (BM-MSC) derived from Sprague Dawley rats and to assess the effect of Cissus quadrangularis Salisb. extract on their maturation into bone cells. The BM-MSC were divided into three groups: (a) BM-MSCs + osteoblast cell growth basal medium as the positive control; (b) BM-MSCs + Dulbecco's modified eagle's medium (DMEM) + 0.3 mg/mL methanol extract of C. quadrangularis as methanol group; and (c) BM-MSC + DMEM + 0.3 mg/mL ethyl acetate extract of C. quadrangularis as ethyl acetate group. A relative quantification approach using was used to analyze the expression of the alp (alkaline phosphatase) gene, with the beta-actin gene was used to normalize the expression of the alp gene. The intra-assay variation was calculated to validate the RT-qPCR data. Our study found that the intra-assay variation value was acceptable, with most of the coefficients of variability (CV) value <5. Ethyl acetate solvent outperformed methanol solvent in extracting the active compound C. quadrangularis. In the ethyl acetate extract group, the expression of the alp gene increased three times compared to the positive control. In methanol extract group, the expression of alp gene was lower six times compared to positive control. This study suggests that C. quadrangularis extracts using ethyl acetate could induce the maturation of BM-MSCs. However, further studies are warrant to confirm this effect using different indicators.
退行性疾病,如骨质疏松症,可以通过干细胞治疗。本研究旨在鉴定骨髓间充质干细胞(BM-MSC)的基因表达,评估菝葜提取物对其向成骨细胞成熟的影响。将 BM-MSC 分为三组:(a)BM-MSCs+成骨细胞生长基础培养基作为阳性对照;(b)BM-MSCs+DMEM+0.3mg/mL菝葜甲醇提取物作为甲醇组;(c)BM-MSCs+DMEM+0.3mg/mL菝葜乙酸乙酯提取物作为乙酸乙酯组。使用相对定量方法分析 alp(碱性磷酸酶)基因的表达,以β-肌动蛋白基因标准化 alp 基因的表达。计算组内变异以验证 RT-qPCR 数据。我们的研究发现,组内变异值是可以接受的,大多数变异系数(CV)值<5。乙酸乙酯溶剂在提取菝葜的活性化合物方面优于甲醇溶剂。在乙酸乙酯提取物组中,与阳性对照相比,alp 基因的表达增加了三倍。在甲醇提取物组中,alp 基因的表达比阳性对照低六倍。本研究表明,使用乙酸乙酯的菝葜提取物可以诱导 BM-MSCs 的成熟。然而,需要进一步的研究来使用不同的指标来确认这种效果。