Effects Of YIGU CAPSULE Drug-containing Serum On IGF-I And NO Secretion Of Osteoblasts And Osteoclasts Apoptosis In Vitro

Objective:The effects of YIGU CAPSULE drug-containing serum on IGF-I and NO secretion of osteoblasts and osteoclasts apoptosis in vitro was studied in order to further explore the mechanism of it preventing and treating osteoporosis.Methods:(1) The 12-month old female Sprague-daweley rats were divided into randomly four groups (YIGU CAPSUL group and saline group), the drug-containing serum and control serum were prepared. (2) The mew-born Sprague-daweley rat Osteoblasts were cultured with different YIGU CAPSULE drug-containing serum concentration and different exposure time, MTT method was used to observe proliferation of Osteoblasts. (3) RT-PCR method was used to measure the relative IGF-ImRNA levels and ELISA method was used to measure IGF-I secretion with different exposure time. (4) The mRNA levels of eNOS and iNOS were measured by RT-PCR; the activities of eNOS and iNOS were measured by test kit with different exposure time. NO secretion of osteoblasts was observed, too. (5) The newborn Sprague-daweley rat Osteoclasts was cultured with different YIGU CAPSULE drug-containing serum concentration and different culture time, TRACP activity was measured by test kit; the survival osteoclasts count was observed under reverse microscope; the percentage of osteoclasts apoptosis were observed under fluorescence microscope after Acridine orange staining.Results:(1) Proliferation of Osteoblasts was more than the control groups respectively after 48, 72 and 96hours(P<0.01); (2) The relative IGF-ImRNA levels and IGF-I proteinexpressions were higher than the control groups respectively after 48, 72 and 96hours(P<0.01 or P<0.05); (3) The mRNA levels and the activities of eNOS and iNOS and NO secretion of osteoblasts were higher than the control groups respectively after 48, 72 and 96hours(P<0.01 or P<0.05); (4) TRACP activity and the percentage of osteoclasts apoptosis were higher than the control groups respectively after 48, 72 and 96hours(P<0.01) and the survival osteoclasts count was less than the control groups respectively after 48, 72 and 96hours(P<0.01).Conclusions:It was suggested YIGU CAPSULE Drug-containing serum could promote proliferation, IGF-I mRNA, IGF-I protein, NOS mRNA and NOS activities expressions and NO secretion of Osteoblasts, and could induce apoptosis and inhibit TRACP activities of osteoclasts. These consults may be parts of the mechanism of YIGU CAPSULE preventing and treating osteoporosis.