Effect Of Serum Containing Benefiting-bone Capsule On The Expression Of IGF-I And TGF-β₁mRNA Of Osteoblast In Co-culture System

Objectives:To study the effects of rat sera-containing Bone-Benefiting Capsule (BBC) on IGF-Ⅰ and TGF-β1 in osteoblasts-osteoclasts co-culture system and its mechanism of action in prevention and treatment of osteoporosis.Methods:1) The osteoblasts-osteoclasts co-culture system was established with agarose gel and microporous membrane (0.45 μm), which prevents the two kinds of cells from contacting directly but allows the free movement of the media.2) Forty female SD rats, aged 10 months, were randomly divided into 4 groups including N.S., high dosage, middle dosage and low dosage with BBC to prepare the drug-containing sera.3) The effect of the rats' sera on osteoblasts' proliferation at different concentrations was determined by the MTT method, which is to determine the best dosage added to the co-culture.4) ELISA was used to determine the effect of the drug-containing sera on expression of IGF-I in the co-culture system at 48h, 72h and 96h.5) FQ-PCR was used to determine the effect of the drug-containing sera on expression of the TGF-β1 mRNA in the co-culture system.Results:1) We demonstrated that the protein can disperse from one side of the microporous membrane to another, indicating that medium can exchange in the co-culture system.2) The drug-containing sera of the high dosage at 25% and 30% obviously promoted osteoblast proliferation, which is statistically significant (P<0.01).3) The levels of IGF-I in the drug-containing sera group were significantlyhigher than that of the control group at 48h, 72h and 96h, which is statistically significant (PO.01).4) The expression of TGF-pimRNA of osteoblast in the drug-containing group was obviously higher than that in the control group, which is statistically significantConclusions:Rat sera-containing BBC can increase levels of IGF-1 and TGF-pl in the co-culture system, which promotes osteoblast proliferation and accelerates osteoclast apoptosis. This could be the mechanism of action of BBC, a drug useful in prevention and treatment of osteoporosis.