| BackgroundWith the rapid development of our country’s economy and medical, theliving conditions of the people are steadily improving, and the proportion ofolder people has increased significantly. China is entering the aging society, andthe aging disease are becoming more and more apparent, such as osteoporosis,which often occurs in the elderly people, causing the lower bone mass and thedeterioration of bone microstructure. In the youth, bone metabolism is balanced,but the balance is broken in the elderly, so with the decreasing of the bone massand bone strength, the risk of fractures in the elderly increased, It becomeshotspot for the scholars to find the etiology and treatment of osteoporosis.Bone marrow mesenchymal stem cells (BMMSCs) was found in the bonemarrow, and has the ability of multi-directional differentiation which attractedthe attention of scholars.If too much BMMSCs were to differentiate into fatcells, the number of BMMSCs to osteoblast differentiation would be decreased,which could lead to osteogenesis defects resulting in osteoporosis.But on thecontrary, it provides a new direction for the treatment of osteoporosis. It is always a hot spot for researchers that how to reduce the side effects inthe process of treatment. In recent years, the scholars focus on a selectiveestrogen receptor modulators—Ginkgo biloba extract (Ginkgo biloba, extract,GBE), whitch can promote the osteogenic differentiation of BMMSCs in vitrobesides antioxidant activities, regulating vascular active, scavenging freeradicals, anti platelet activating factor, neural protection, increase blood flowpharmacological action, but adipogenic differentiation ability of the BMMSCshas not been reported in the presence of GBE. So the purpose of this study wasto investigate the effects of different concentrations of GBE on the adipogenicdifferentiation ability of BMMSCs, illuminating the significance of GBE onosteoporosis treatment and prevention in the cell level.ObjectiveTo study the effect of different concentration of Ginkgo biloba extract(GBE) on adipogenic differentiation ability of rat bone marrow mesenchymalstem cells (BMMSCs) and illuminating the significance of GBE on osteoporosistreatment and prevention in the cell level.Content:1．Add50,100,150,200and400mg L-1GBE and adipogenic liquid to the purified P3BMMSCs (50,100,150,200,400mg L-1GBE group), with no drugcontrol group, witch were cultured for7days.Observe the expression ofBMMSCs on morphology and surface antigen CD44, CD105and CD34.2．Observe oil red O staining of BMMSCs in the formation of lipid droplet;analysis the quantitative of the absorbance value (A) to observe the adipogenicability of BMMSCs.3．Use RT-PCR method to detect the expression level of the adipose tissuefatty acid binding protein (AP2) and peroxisome proliferator activated receptor(PPAR) subtypes (PPAR-γ) mRNA.Result1.The morphological detection： After3days cultured BMMSCs cellswere spindle shaped in vitro.After7days the cells were swirling growth andarranging in the same direction.The surface antigen staining showed that theexpression of CD44and CD105was active, and the expression of CD34wasnegative. The cells were proved to be the BMMSCs.2.Oil red O staining for7days and a large number of lipid droplet formedunder a microscope, and with the increase of concentration of GBE the numberof lipid droplets reduced; absorbance detection results showed that comparedwith the control group, A510decreased gradually with the increase of theconcentration of GBE (P<0.05). 3. The results of RT-PCR showed that compared with the control groupwith the increase of the concentration of GBE the expression of PPAR-γandAP2mRNA decreased gradually (P<0.05).ConclusionGBE can inhibit the adipogenic differentiation ability of BMMSCs in vitro,and the inhibition increases with the concentration of GBE increasing. |
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