Ra ostcobasts were isolated from the 21-day fetal rat calvchas. The cells were grown in DMEM Plus 10% FBS, and were treated for 24 h. with 10 μmol/L TPEN or 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+. Apoptos...Ra ostcobasts were isolated from the 21-day fetal rat calvchas. The cells were grown in DMEM Plus 10% FBS, and were treated for 24 h. with 10 μmol/L TPEN or 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+. Apoptosis of osteoblasts were measured by fiow cytometry, electron microscopy and DNA fragmentation analyzed by gel elecmphoresis. In addition, IP3 production and PKC activity were measmed in ordr to show whether they are involved in apoptosis in osteoblast induced by alnc deficiency. The results showed that 10 μmol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells ed with 10μmol/L TPEN supplemented with 10 μmol/L Zn2+ showed no apoptotic changs in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supPlemented with Zn2+ simulaneosly and the untreated cells. It can be inferred that apoptosis induced by ainc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3.展开更多
<正>Objective To investigate the combined effects of fluoride(Na F)and arsenate(Na As O2)exposure on proliferation,differentiation and bata-catenin expression in SD rat osteoblasts.Methods Osteoblasts were isola...<正>Objective To investigate the combined effects of fluoride(Na F)and arsenate(Na As O2)exposure on proliferation,differentiation and bata-catenin expression in SD rat osteoblasts.Methods Osteoblasts were isolated from calvarias of twelve SD rats born in 1~3 days and cul-展开更多
Background Calcitonin gene-related peptide （CGRP）, a sensory neuropeptide, affects osteoblast proliferation and bone formation. However, the mechanisms are not fully understood. Monocyte chemoattractant protein-1 （...Background Calcitonin gene-related peptide （CGRP）, a sensory neuropeptide, affects osteoblast proliferation and bone formation. However, the mechanisms are not fully understood. Monocyte chemoattractant protein-1 （MCP-1） is a chemokine that stimulates the migration of monocytes and plays important roles in regulating bone remolding during fracture repair. In this study, we investigated the effects of CGRP on proliferation and MCP-1 expression in cultured rat osteoblasts. Methods Primary rat osteoblasts were isolated from fetal rats calvariae. Cells were exposed to gradient concentrations （10^-9 to 10^-7 mol/L） of CGRP. Protein and mRNA levels of MCP-1 were quantified by Western blotting and semiquantitative reverse transcdption-polymerase chain reaction, respectively. The protein level of MCP-1 was investigated and compared in cell culture media by enzyme linked immunosorbent assay （ELISA）. Phospho-extracellular signal-regulated kinase （ERK） expression was detected by Western blotting. Cell proliferative activity was measured by 3-（4,5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide （MTT） and BrdU assay. The effects of MAPK/ERK kinase （MEK）-inhibitor U0126 on CGRP-induced MCP-1 expression in primary rat osteoblasts were examined. Results CGRP effectively enhanced primary rat osteoblast proliferation and led to significant increases in the expression of MCP-1 mRNA and protein in time- and dose-dependent manners. CGRP activated the ERK pathway. Pretreatment of cultured rat osteoblasts with MEK inhibitor U0126 resulted in dose-dependent inhibitions of CGRP-induced MCP-1 mRNA and protein levels. Thus, CGRP promoted cell proliferation and stimulated MCP-1 expression in cultured rat osteoblasts. Conclusion These studies document novel links between CGRP and MCP-1 and illuminate the effects of CGRP in regulating bone remodeling.展开更多
The regulation of cellular differentiation by progesterone in fetal rat calvarial osteoblasts was investigated. Our results showed that cells cultured in the presence of progesterone had a 7% increase in the alkaline ...The regulation of cellular differentiation by progesterone in fetal rat calvarial osteoblasts was investigated. Our results showed that cells cultured in the presence of progesterone had a 7% increase in the alkaline phosphatase activity when compared to untreated cells. The concentration of osteocalcin in the conditioned medium from progesterone treated osteoblasts was 28% higher than that of untreated controls. In addition,administration of progesterone significantly enhanced the number and area of bone nodules. In conclusion, progesterone stimulates the differentiation of fetal rat calvarial osteoblastic cells in vitro展开更多
Objective: To observe the effects of different gravitational environments on release of prostaglandin E2 in rat calvarial osteoblasts induced by fluid shear stress (FSS) so that to investigate the influence of differe...Objective: To observe the effects of different gravitational environments on release of prostaglandin E2 in rat calvarial osteoblasts induced by fluid shear stress (FSS) so that to investigate the influence of different gravity on mechanotransduction in osteoblasts. Methods: Osteoblasts were isolated from neonatal rat calvariae and then were set to three groups. Each was cultured in one gravitational environment; 1G terrestrial gravitational environment (control), simulated weightlessness achieved by using clinostat and 3G gravitational environment achieved by using centrifuge for 60 h, then osteoblasts were treated with 0. 5 Pa or 1. 5 Pa FSS in a flow chamber for 1 h. The release of PGE2 in osteoblasts was determined. Results: In 1G gravitational environment, the release of PGE2 was significantly increased along with the sustaining of FSS treatments (P<0. 01), but there was no remarkable difference between the responses to 0. 5 Pa FSS and 1. 5 Pa FSS (P>0. 05). While in simulated weightlessness展开更多
文摘Ra ostcobasts were isolated from the 21-day fetal rat calvchas. The cells were grown in DMEM Plus 10% FBS, and were treated for 24 h. with 10 μmol/L TPEN or 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+. Apoptosis of osteoblasts were measured by fiow cytometry, electron microscopy and DNA fragmentation analyzed by gel elecmphoresis. In addition, IP3 production and PKC activity were measmed in ordr to show whether they are involved in apoptosis in osteoblast induced by alnc deficiency. The results showed that 10 μmol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells ed with 10μmol/L TPEN supplemented with 10 μmol/L Zn2+ showed no apoptotic changs in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supPlemented with Zn2+ simulaneosly and the untreated cells. It can be inferred that apoptosis induced by ainc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3.
文摘<正>Objective To investigate the combined effects of fluoride(Na F)and arsenate(Na As O2)exposure on proliferation,differentiation and bata-catenin expression in SD rat osteoblasts.Methods Osteoblasts were isolated from calvarias of twelve SD rats born in 1~3 days and cul-
文摘Background Calcitonin gene-related peptide （CGRP）, a sensory neuropeptide, affects osteoblast proliferation and bone formation. However, the mechanisms are not fully understood. Monocyte chemoattractant protein-1 （MCP-1） is a chemokine that stimulates the migration of monocytes and plays important roles in regulating bone remolding during fracture repair. In this study, we investigated the effects of CGRP on proliferation and MCP-1 expression in cultured rat osteoblasts. Methods Primary rat osteoblasts were isolated from fetal rats calvariae. Cells were exposed to gradient concentrations （10^-9 to 10^-7 mol/L） of CGRP. Protein and mRNA levels of MCP-1 were quantified by Western blotting and semiquantitative reverse transcdption-polymerase chain reaction, respectively. The protein level of MCP-1 was investigated and compared in cell culture media by enzyme linked immunosorbent assay （ELISA）. Phospho-extracellular signal-regulated kinase （ERK） expression was detected by Western blotting. Cell proliferative activity was measured by 3-（4,5-dimethylthiazol-2-yl）-2, 5-diphenyl tetrazolium bromide （MTT） and BrdU assay. The effects of MAPK/ERK kinase （MEK）-inhibitor U0126 on CGRP-induced MCP-1 expression in primary rat osteoblasts were examined. Results CGRP effectively enhanced primary rat osteoblast proliferation and led to significant increases in the expression of MCP-1 mRNA and protein in time- and dose-dependent manners. CGRP activated the ERK pathway. Pretreatment of cultured rat osteoblasts with MEK inhibitor U0126 resulted in dose-dependent inhibitions of CGRP-induced MCP-1 mRNA and protein levels. Thus, CGRP promoted cell proliferation and stimulated MCP-1 expression in cultured rat osteoblasts. Conclusion These studies document novel links between CGRP and MCP-1 and illuminate the effects of CGRP in regulating bone remodeling.
文摘The regulation of cellular differentiation by progesterone in fetal rat calvarial osteoblasts was investigated. Our results showed that cells cultured in the presence of progesterone had a 7% increase in the alkaline phosphatase activity when compared to untreated cells. The concentration of osteocalcin in the conditioned medium from progesterone treated osteoblasts was 28% higher than that of untreated controls. In addition,administration of progesterone significantly enhanced the number and area of bone nodules. In conclusion, progesterone stimulates the differentiation of fetal rat calvarial osteoblastic cells in vitro
基金Supported by the Research Foundation for Innovation project of FMMU(No. CX01A012)
文摘Objective: To observe the effects of different gravitational environments on release of prostaglandin E2 in rat calvarial osteoblasts induced by fluid shear stress (FSS) so that to investigate the influence of different gravity on mechanotransduction in osteoblasts. Methods: Osteoblasts were isolated from neonatal rat calvariae and then were set to three groups. Each was cultured in one gravitational environment; 1G terrestrial gravitational environment (control), simulated weightlessness achieved by using clinostat and 3G gravitational environment achieved by using centrifuge for 60 h, then osteoblasts were treated with 0. 5 Pa or 1. 5 Pa FSS in a flow chamber for 1 h. The release of PGE2 in osteoblasts was determined. Results: In 1G gravitational environment, the release of PGE2 was significantly increased along with the sustaining of FSS treatments (P<0. 01), but there was no remarkable difference between the responses to 0. 5 Pa FSS and 1. 5 Pa FSS (P>0. 05). While in simulated weightlessness
