| Objective: To probe into the active mechanism of Morinda officinalis How on invigorate the kidney and building up bones and muscles in molecular biology level. To establish the model of the SD rat's BMSCs cultured with serum of Morinda officinalis How collected by aqueous extract and alcohol extract .To observe proliferation, differentiation to osteoblast and expression of Cbfal of SD rat's marrow stromal cell cultured by aqueous extracting component and alcohol extracting component of Morinda officinalis How. Methods: The serum pharmacology method was used to culture the SD rat's marrow stromal cell. It were divided into 6 groups, A (aqueous extract group), B (alcohol extract group), C (control group), D (classics induce bone formation group), E (aqueous extract + classics induce bone formation group), F (alcohol extract + classics induce bone formation group). To establish the model of the SD rat's BMSCs cultured with serum of Morinda officinalis How collected by aqueous extract and alcohol extract, the A, B and C groups BMSCs were cultured by serum of Morinda officinalis How in different condition, by different dosage, different collection time, different administration time, different concentration sera, the OD value was indicated using MTT method in 72 hours. The flow cytometer was used to measure the content of DNA of A, B, C group in G1, S, G2 stage, which cultured with the same concentration sera in 72hs. The improvement Gomori's calcium-cobalt staning, alizarin Bordeaux staning, Van-Gieson I collagen staining method were used to observe the expressions of ALP, mineralization nodus and type I collagen respectively. The alkaline phosphatase activity and the amount of osteocalcin in medium were measured in different time. Semi-quantitative analysis was used to detect the expression of Cbfal through RT-PCR method. Results: The OD value was higher in 4 days (administration time), Isodose (administration dosage) and 1 hour (collection time) group compared with the control group (P < 0.05), which in different concentration group sequence as: 20% > 10% > 5%. The content of DNA of the aqueous extract group or alcohol extract group compared with thecontrol group in S stage (P < 0.05), which was best in the 20% alcohol extract serum. The intensity of ALP dyeing, alizarin Bordeaux dyeing sequenced as: F>E>D>B>A group (negative in C group), and collagen I dyeing sequenced as: F>E>D>B>A>C group . The alkaline phosphatase activity and the amount of bone glaprotein were higher in A, B, D, E, and F groups compared with the control group at the same time (p<0.05). E and F group were higher than D group at the same time (p<0.05). The Cbfal mRNA was detected 3 days after classic bone formation inducing, and increased gradually, then reached to the peak at 12 days, which decreased in 15 days and 18 days. At 12 days, the expression of the Cbfal mRNA sequenced as: F>E>D>B>A>C, which in A, B, D, E, and F groups were stronger compared with C group (p<0.05).Conclusions:1 The best project which accelerate the proliferation and differentiation of the BMSCs were confirmed as: Isodose, 4 days' administration, killed 1 hour after administrated, deactivation, and 20% medium.2 The Morinda officinalis How collected by aqueous extract and alcohol extract could accelerate the proliferation of the BMSCs, and could induce the BMSCs differentiation, which show the Morinda officinalis How could optimize the Seeded Cells.3 The Morinda officinalis How accelerate the proliferation and differentiation of the BMSCs by improve the activity of ALP in cells, BPG, and type I collagen protein.4 The function of the proliferation and differentiation to osteoblast and increased the expression of Cbfal of BMSCs my be one of the mechanism of the Morinda officinalis How invigorating the kidney, bones and muscles.5 The active site of the Morinda officinalis How may exist in the alcohol extract mostly.
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