The Effect Of Klotho Gene Transfection On The Vitality Of Primary Osteoblast

Objective:To investigate the effect of klotho (KL) gene transfection on the vitality of primary osteoblast, and to reveal the effect of KL via fibroblast growth factor-23 (FGF-23) signal path on bone metabolism.Methods:The primary mesenchymal stem cells (BMSCs) were abstracted from male rats. The morphologies of BMSCs was observed by inverted microscope. After the third generation culture, the specific surface markers of them were identified by flow cytometer. Then BMSCs were cultured using conditioned medium particularly for inducing into osteoblasts for 14-21 days. Alizarin red staining was used to test whether the induction was successful. After successfully induced, the primary osteoblasts were transfected by adeno-associated virus mouse KL gene. The experiment was divided into 5 groups:control group, KL group, KL with BGJ398 (FGFR1 inhibitor) group, adenovirus empty vector group, adenovirus empty vector with BGJ398(FGFR1 inhibitor) group. After treatments, the expression of KL in osteoblasts was detected by RT-PCR, inversed fluorescent microscope and ELISA. The expression of osteoblast discharges:osteopontin (OPN), osteocalcin(OCN) were detected by RT-PCR.Results:BMSCs were successfully abstracted from rats, and also primary osteoblasts were induced. After transfected,the exogenous KL-derived genes were highly expressed (p<0.01).KL could increase the discharge of OCN(P<0.05) and inhibit the discharge of OPN(P<0.05) in osteoblasts when KL-FGF23-FGFR1 pathway was normal.Conclusion:KL can promote the vitality of primary osteoblast probably through KL-FGF23-FGFR1 pathway.