Based On The Cellular Level Of Anti-osteoporosis Medicine Pharmacodynamic Evaluation Of Methods To Establish

Osteoporosis is a leading cause imperiling human health. Seeking effective anti-osteoporosis drugs has been an important issue in medicine. There are many bioactive components that can treat osteoporosis in Traditional Chinese Medicine. Especially, they have less side-effect. Ttraditional Chinese Medicine is becoming more and more important in treating osteoporosis. However, Ttraditional Chinese Medicine have complex components, doctors usually used them according to their previous experience. There were not any people who would study their mechanisms. With the modernization of Traditional Chinese Medicine and advancement of cell-culture technique, we can study the medical effects of Ttraditional Chinese Medicine on osteoporosis in deeper level. This paper aimed to establish a method of pharmacodynamic evaluation of Ttraditional Chinese Medicine on treating osteoporosis based on cellular level.Firstly, we cultured osteoblasts and osteoclasts in vitro. Enzyme-digestion was used to obtain osteoblasts from newborn SD rats. And alkaline phosphatase staining was used to identify osteoblasts. Osteoclasts were generated by inducing bone marrow cells of SD male rat with 1,25(OH)₂D₃. Osteoclasts were identified through tartrate-resistant acid phosphatase (TRAP) staining. At last, we observed both the F-actin of osteoblasts and osteoclasts through Laser Scanning Confocal Microscope after double-labeling with both Bodipy FL Phallacidin and PI.Secondly, we used traditional pharmacological methods to make serum containing drugs and improved it. In traditional methods, drugs were given directly to rats. Later, blood was sampled and serum containing drugs was made. As we all know, there are quite a few patients suffering osteoporosis that are women in postmenopause. The decline of estrogen has a negative effect on osteoporosis. In this paper, before drugs were given to female rats, we extirpated their ovaries to get rid of estrogen in serum. Therefore, we can study the effects of drugs components in serum on cell function.Thirdly, we established pharmacodynamic evaluation of Chinese Medicine on bone formation of osteoblasts cultured in vitro. WST-1 was used to examine the proliferation ofosteoblasts. Alizarin dying was used to exmine mineralization of osteoblasts. The activity of ALP and secretion of IGF-1 was measured with spectrophotometer. At last, We observed the effects of serum containing 10% Gui-Lu compound on the function of osteoblasts. We found that high doses of serum containing Gui-Lu compound had obvious effects on stimulating the multiplication and proliferation of osteoblasts(P<0.01). We found that middle doses could enhance the activity of ALP of osteoblasts. High doses could increase mineralization and secretion of IGF-1 of osteoblasts. This suggested that Gui-Lu compound could obviously promote bone formation of osteoblasts.Last, we established pharmacodynamic evaluation of Chinese medicine on bone asorption of osteoclasts cultured in vitro. We used Electron Scanning Microscope to observe bone pits of osteoclasts and TRAP(+) cells with more than two nuclei and TRAP activity of osteoclasts were examined. At the same time, we observed the effects of serum containing Gui-Lu compound on the function of osteoclasts. We found that middle doses obviously inhibited the formation of bone pits. Moreover, we found that middle doses could oberviously inhibit the formation of osteoclast and TRAP (+) cells. TRAP activity of osteoclasts was also lowered clearly. This study indicated that Gui-Lu compound can inhibit the asorption of osteoclasts.From the above results, we established pharmacodynamic evaluation of anti-osteoporosis Chinese Medicine. And we used Gui-Lu compound to validate it and found that the mechanisms of Gui-Lu compound on treating osteoporosis may be related to the following two aspects: (1) Increase bone formation of osteoblasts;(2) Inhibit bone asorption of osteoclasts. This paper suggested that Gui-Lu compound had obvious positive effects on treating osteoporosis. And the feasibility of this method was further validated.