Application of rhoifolin in preparation of drug for preventing and treating osteolytic diseases
A kind of rhodol glycoside and osteolysis technology, which is applied in bone diseases, drug combinations, pharmaceutical formulations, etc., and can solve problems such as prevention and treatment of orthopedic diseases that have not been seen
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[0019] 1. Materials and methods
[0020] 1.1 Materials
[0021] Medium: DMEM, a-MEM, and fetal bovine serum (FBS) purchased from Gibco-BRL, USA.
[0022] Reagent: Rhodorin (C27H30O14, molecular weight 578.52) (Chengdu). Antibodies: IkBa, ERK, p-ERK, JNK, p-JNK, p38, p-p38, NFATc1, c-Fos), secondary antibody, b-actin (USA). M-CSF, RANKL (USA). Hydroxyapatite coated culture plate (Beijing), luciferase lysate buffer, DTT solution (USA), titanium particles (USA) and other materials.
[0023] 1.2 Osteoclast differentiation test
[0024] Bone marrow macrophages (BMMs) were harvested from 8-week-old C57BL / 6J mice, and the bone marrow was flushed from the femur and tibia using a small needle. All cells were then cultured overnight in complete α-MEM medium. Then place the BMMs in 8x10 3 The density per well was seeded in 96-well plates and cultured in α-MEM for 24 hours to adhere. For osteoclast differentiation and drug assays, cells were cultured with 100 ng / mL recombinant RAN...
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