Proliferation, Differentiation And Apoptosis Molecular Biology Study On The Adjust Mechanism Of SD Rat's Marrow Stromal Cell By Sthenia Skeleton Herbs Mixture In Vitro

Object: We use SD rat's marrow stromal cell cultured in vitro. To find out the principle of treatment on osteoporosis by Sthenia Skeleton Herbs Mixture and observe the proliferation, differentiation and apoptosis molecular biology adjust mechanism. Method: Marrow stromal cell were separated from the extremity of 30-day SD rat extremity in vitro. Using MTT method to measure the effect on marrow stromal cell proliferation by Sthenia Skeleton Herbs Mixture. Applying morphologic, cell ultrastructure observation, alkaline phosphatase specific activity, inner-cell osteocalcin, |3 transforming growth factor 1 gene expression and so on to investigate the level of marrow stromal cell transfer into osteoblast. Adopting morphologic, cell ultrastructure observation, apoptosis cell cycle and mitochondrion membrane potential changes, bcl-2> bax protein gene expression and so on to evaluate the effect of Sthenia Skeleton Herbs Mixture on the course of cell apoptosis. Result: A certain range concentration of Sthenia Skeleton Herbs Mixture has the significant effect to accelerate the marrow stromal cell proliferation in vitro. It can also hasten the cell excreting alkaline phosphatase and osteocalcin. Reinforce the activity of recombinated human bone morphogenetic protein 2. Facilitate the cell express β transforming growth factor 1. These effect are the same as Miacalcic. The earlier use of Sthenia Skeleton Herbs Mixture can decreasing the cells that are blocked at GO/G1. Promoting it compose DNA. The expression of bcl-2 is higher in Sthenia Skeleton Herbs Mixture group than that of induced group, and the expression of bax is lower in Sthenia Skeleton Herbs Mixture group than that of induced group. Mitochondrion membrane potential descent very significant in induction group than in Sthenia Skeleton Herbs Mixture group. Conclusion: There are three principle of treatment on osteoporosis by Sthenia Skeleton Herbs Mixture. First, Sthenia Skeleton Herbs Mixture contains such component that can directly stimulate the proliferation of marrow stromal cell, which can rise the amount of transforming to osteoblast. Consequently accelerated the bone formation. Second, it can reinforce the activity of recombinated human bone morphogenetic protein2. Hastening the transforming marrow stromal cell express β transforming growth factor 1 to excreting collagen I .Thus benefit the deposition of calcium salts. It can also hastening the induced marrow stromal cell by recombinated human bone morphogenetic protein 2 excrete alkaline phosphatase and osteocalcin, that can accelerate the transforming into osteoblasts. Sequentially accelerate the deposition of calcium and Phosphorus on the bone surface. Third, the earlier use of Sthenia Skeleton Herbs Mixture can decrease the amount of apoptosis cell induced by all trans retinoic acid. Thus promoted cell mitosis and restrain apoptosis. It can also act as protection role with regard to bcl-2 that located on the mitochondrion membrane. Preventing bax protein from cell-plasm to mitochondrion membrane to take the advantage of bcl~2 in formating bcl-2 & bax homology dimer, preventing the opening of permeability transforming bore to prevent the destruction of mitochondrion membrane potential and biosynthesis by mitochondrion release apoptosis induced factor to reach the objective of restrain apoptosis.