| Osteoprotegerin is an inhibitory factor of Osteoclasts. It has been the potential treatment for osteoporosis. OPG is a key factor acting as a negative regulator against osteoclasts' differentiation and activation, through competitive inhibition of RANKL's binding to its membrane receptor - Receptor Activator of Nuclear Factor-κB (RANK). In that way, OPG interferes with the signal pathway of Osteoclasts and their ancestor, and showing excellent effect of inhibiting bone resorption function not only in animal experiments but also in clinical application.Although it can inhibit Osteoclasts, OPG is expensive (For example, the market price of OPG expressed by yeast is $ 4.2/mg). It will cost a lot of monkey for the patients if they use OPG for a long period of time. On the other hand, abnormal allery immunologic reaction could be found in those patients who use the OPG for a long time. For lighting the economic load of patients and lowering the side effect, we reconstructed a new OPG-Fc yeast expression system and try to combine OPG and some other drugs to enhance the effect of inhibiting the Osteoclasts.In this study, we used the methylotrophic yeast Pichia pastoris-SMD1168 to rebuild SMD1168/OPG-Fc system. Comparing with the former system we had already built--X33/OPG-Fc, SMD1168 grow slower but express less non-target protein. And the OPG-Fc produced by both SMD1168 and X33 has inhibited Osteoclasts function.In the experiment, we accessed combined effect of rhOPG-Fc and Alendronate (ALN) on osteoclasts. ALN can inhibite farnesyl diphosphate synthase of Mevalonate Pathway of osteoclast, and induce these cells' apoptosis. The results showed that combination of rhOPG-FC and ALN had the great inhibition effect on mature osteoclasts and their precursor cells' differentiation. And half dose of OPG + ALN had greater effect on inhibiting the Osteoclast than using OPG or ALN only. In an other experiment, OPG + ALN also showed the... |
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