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Experimental Study of the Proliferation and Osteogenic Differention Effects of Psoralen on Bone Mesenchymal Stem Cells from Rats

Author: LinQiang
Tutor: ZhangRongHua
School: Jinan University
Course: Traditional Chinese Medicine
Keywords: psoralen bone mesenchymal stem cells osteoblast proliferation
CLC: R285.5
Type: Master's thesis
Year: 2009
Downloads: 98
Quote: 0
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Abstract


Objective:To explore the mechanism of Psoralen on invigorating the kidney and building up bones on different levels To observe the effects on proliferation,differentiation to osteoblast of BMSCs cultured by various concentrations of Psoralen.To investigate the expression of TGF-β1 mRNA in induced system with Psoralen on molecular biology level.Methods:1.BMSCs from SD rats were isolated and purified by using attachment method,then identified according to morphology and induced differentiation potentiality.2.MTT method was used to investigate the proliferation effects different concentrations Psoralen on BMSCs in SD rats.3.ALP-positive cell counts by the improvement Gomori’s calcium-cobalt staining was explored to observe.Osteoblastic differentiation potentiality of BMSCs induced by different concentration Psoralen.4.Groups were divided:blank group,Psoralen group and classics group.The ALP activity and the number of mineralized nodesls were measured to indicate the osteoblast function of different groups.5.Groups were divided:blank group,Psoralen group and classics group.Semi-quantitative analysis was used to detect the expression of TGF-β1 mRNA during osteodifferen-tiation through RT-PCR method.Result:1.Primary cultured BMSCs adhered to plastic surface within 24 hours and reached confluence within 10～12 days.To be conditioned induced,BMSCs were successfully differentiated into osteoblast and adipocyte.And confirmed that there was expression of CD29 and CD44,and no expression of CD34 and CD45 on the surface of MSCs by flow cytometry （FCM）.2.OD values of MTT shows Psoralen groups of 15μmol/L、10μmol/L、5μmol/L were higher than blank groups,significantly（P＜0.05）,especially 10μmol/L. 3.Results of ALP staining for ALP-positive cell counts showed Psoralen of 5μmol/L、10μmol/L、15μmol/L addition had a definit induction to osteoblast differentiation compared with the blank with significant difference（P＜0.05）,especially 15μmol/L.4.There were significant differences in Psoralen group,classics group and blank group in the average number of mineralized nodes in BMSCs 21 days.（P＜0.05）5.There were significant differences between Psoralen group and blank group on the expression of the TGF-β1 mRNA.（P＜0.05）.ConclusionBMSCs can be isolated from rat bone marrow by the method of different attachment.The obtained BMSCs were purified and had great capacity to proliferate.Psoralen could accelerate the proliferation and induce the differentiation to osteoblast of BMSCs.The mechanism of psoralen on preventing and curing osteoporosis may be related to accelerating the proliferation as well as inducing the osteodifferentiation by improving the activity of ALP and enhancing the expression of TGF-β1 mRNA.

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