Effects Of Osthole On Proliferation And Differentiation Of Osteoblasts Based On Endoplasmic Reticulum Stress And Wnt/?-Catenin Signaling Pathway

BACKGROUND:Osteoporosis is a metabolic bone disease caused by bone resorption-bone formation coupling imbalance,and its treatment focuses on the recovery of bone metabolism.Osthol has been shown effective with osteoporosis,but its mechanism remains unclear.OBJECTIVE:To observe the effect of osthol on the proliferation and differentiation of osteoblasts in rats,and to explore the mechanism of osthol of anti-osteoporosis effect.METHODS:In this study,a large number of high-purity primary osteoblasts were obtained from the skull of neonatal rats by modified enzyme digestion and repeated adherence purification method.The osteoblasts were obtained for follow-up experiments and observed by light microscopy,Alkaline phosphatase staining,alizarin red mineralization nodular staining to successfully identify osteoblasts.The author finally selected 10^-6M,10^-5M,10^-4M,respectively,as low,medium and high concentrations of osthol group.In addition,10^-8M beta-estradiol as well as solvent control group was established according to the literature.Finally a blank group was set up to exclude solvent interference on osteoblasts.RESULTS:10^-4M osthol could inhibit the proliferation of osteoblasts.10^-5M osthol could inhibit osteoblast proliferation after 72 hours of administration.10^-4M osthol could promote osteoblast expression,10^-4M osthol group began to increase the alkaline phosphatase in osteoblasts after 48 hours of administration.10^-4M osthol could promote the secretion of osteocalcin in osteoblasts.The concentration of ochratins could promote Osteoblast mineralized nodules formed,of which 10^-4M ostricans the strongest effect;the concentration of snake elements can reduce the expression of GRP78,especially 10^-4M concentration is most significant,and PDI expression and osthol The expression of CHOP was increased only in the 10^-4M concentration group.The expression of Wntl protein in 10^-4M and 10^-5M group was increased,while that of ?-catenin was only 10^-4M group increased.CONCLUSION:Osthol can promote the proliferation and differentiation of ?-catenin-mediated osteoblasts by increasing the level of Wntl protein.However,the ability of exocytosis of high concentration osthol is stronger than that of osteoblast Apoptosis,which greatly inhibited osteoblast proliferation,osthol to stimulate osteogenesis of the best concentration of 5 × 10-5M?10-4M between.This study confirmed the role of osthol in promoting bone formation and put forward its mechanism,which laid the foundation for the development of new drug with ochratin as active ingredient.