中国骨伤
中國骨傷
중국골상
CHINA JOURNAL OF ORTHOPAEDICS AND TRAUMATOLOGY
2015年
5期
446-449
,共4页
李军%王云%鲍小明%卫鹏斌%张民
李軍%王雲%鮑小明%衛鵬斌%張民
리군%왕운%포소명%위붕빈%장민
重组人骨形态发生蛋白-2%骨质疏松%血管内皮因子%骨髓基质干细胞
重組人骨形態髮生蛋白-2%骨質疏鬆%血管內皮因子%骨髓基質榦細胞
중조인골형태발생단백-2%골질소송%혈관내피인자%골수기질간세포
Recobinant human bone morphogenetic protein-2(rhBMP-2)%Osteoporosis%Vascular endothelial growth factors(VEGF)%Bone marrow stromal stem cells(BMSCs)
目的：观察骨形态发生蛋白-2对骨质疏松时骨髓基质干细胞（BMSCs）体外成骨及成骨因子VEGF表达的影响，为骨质疏松证的防治提供新的方法。方法：将20只6月龄，体重（300±20）g雌性SD大鼠双侧卵巢切除，术后3个月利用双能X线骨密度仪测量大鼠全身骨密度并与术前比较，确保造模成功，并运用全骨髓贴壁法培养骨质疏松大鼠BMSCs，倒置相差显微镜下观察BMSCs形态。随机把骨质疏松大鼠BMSCs第2代（p2）细胞分成实验组和对照组，分别加入完全培养基（含rhBMP-2）、成骨诱导液进行成骨诱导。2周后茜素红染色法检测各组细胞钙结节的形成，酶标仪测定碱性磷酸酶活性及RT-PCR法检测VEGF的表达量。结果：（1）大鼠全身骨密度：手术前后大鼠全身骨密度分别为（0.179±0.007），（0.158±0.006）g/cm2，差异有统计学意义（t=4.180，P<0.05）。（2）茜素红染色：BMSCs（P2）成骨诱导2周后实验组染色效果明显强与对照组。（3）碱性磷酸酶活性：BMSCs（P2）成骨诱导2周后碱性磷酸酶活性实验组明显高于对照组，分别为（15.62±1.27），（8.62±0.93）μg/prot，差异有统计学意义（t=7.709，P<0.01）。（4）BMSCs（P2）成骨诱导2周后VEGF表达：实验组明显高于对照组，分别为3.723±0.143，0.950±0.072，差异有统计学意义（t=29.462，P<0.01）。结论：rhBMP-2能提高去卵巢骨质疏松大鼠BMSCs的体外成骨能力，可促进成骨因子VEGF的表达，调控VEGF的表达可能是骨形态发生蛋白-2参与骨代谢的机制之一。
目的：觀察骨形態髮生蛋白-2對骨質疏鬆時骨髓基質榦細胞（BMSCs）體外成骨及成骨因子VEGF錶達的影響，為骨質疏鬆證的防治提供新的方法。方法：將20隻6月齡，體重（300±20）g雌性SD大鼠雙側卵巢切除，術後3箇月利用雙能X線骨密度儀測量大鼠全身骨密度併與術前比較，確保造模成功，併運用全骨髓貼壁法培養骨質疏鬆大鼠BMSCs，倒置相差顯微鏡下觀察BMSCs形態。隨機把骨質疏鬆大鼠BMSCs第2代（p2）細胞分成實驗組和對照組，分彆加入完全培養基（含rhBMP-2）、成骨誘導液進行成骨誘導。2週後茜素紅染色法檢測各組細胞鈣結節的形成，酶標儀測定堿性燐痠酶活性及RT-PCR法檢測VEGF的錶達量。結果：（1）大鼠全身骨密度：手術前後大鼠全身骨密度分彆為（0.179±0.007），（0.158±0.006）g/cm2，差異有統計學意義（t=4.180，P<0.05）。（2）茜素紅染色：BMSCs（P2）成骨誘導2週後實驗組染色效果明顯彊與對照組。（3）堿性燐痠酶活性：BMSCs（P2）成骨誘導2週後堿性燐痠酶活性實驗組明顯高于對照組，分彆為（15.62±1.27），（8.62±0.93）μg/prot，差異有統計學意義（t=7.709，P<0.01）。（4）BMSCs（P2）成骨誘導2週後VEGF錶達：實驗組明顯高于對照組，分彆為3.723±0.143，0.950±0.072，差異有統計學意義（t=29.462，P<0.01）。結論：rhBMP-2能提高去卵巢骨質疏鬆大鼠BMSCs的體外成骨能力，可促進成骨因子VEGF的錶達，調控VEGF的錶達可能是骨形態髮生蛋白-2參與骨代謝的機製之一。
목적：관찰골형태발생단백-2대골질소송시골수기질간세포（BMSCs）체외성골급성골인자VEGF표체적영향，위골질소송증적방치제공신적방법。방법：장20지6월령，체중（300±20）g자성SD대서쌍측란소절제，술후3개월이용쌍능X선골밀도의측량대서전신골밀도병여술전비교，학보조모성공，병운용전골수첩벽법배양골질소송대서BMSCs，도치상차현미경하관찰BMSCs형태。수궤파골질소송대서BMSCs제2대（p2）세포분성실험조화대조조，분별가입완전배양기（함rhBMP-2）、성골유도액진행성골유도。2주후천소홍염색법검측각조세포개결절적형성，매표의측정감성린산매활성급RT-PCR법검측VEGF적표체량。결과：（1）대서전신골밀도：수술전후대서전신골밀도분별위（0.179±0.007），（0.158±0.006）g/cm2，차이유통계학의의（t=4.180，P<0.05）。（2）천소홍염색：BMSCs（P2）성골유도2주후실험조염색효과명현강여대조조。（3）감성린산매활성：BMSCs（P2）성골유도2주후감성린산매활성실험조명현고우대조조，분별위（15.62±1.27），（8.62±0.93）μg/prot，차이유통계학의의（t=7.709，P<0.01）。（4）BMSCs（P2）성골유도2주후VEGF표체：실험조명현고우대조조，분별위3.723±0.143，0.950±0.072，차이유통계학의의（t=29.462，P<0.01）。결론：rhBMP-2능제고거란소골질소송대서BMSCs적체외성골능력，가촉진성골인자VEGF적표체，조공VEGF적표체가능시골형태발생단백-2삼여골대사적궤제지일。
Objective:To observe the impact of bone morphogenetic protein-2(rhBMP-2)on bone marrow stromal cells (BMSCs)osteogenesis in vitro and vascular endothelial growth factors(VEGF)expression in bone osteoporotic to prevent and treat the osteoporosis. Methods:Twenty 6 month old female SD rats weighted(300±20)g underwent bilateral ovariecto?mized. At 3 months after operation,dual energy X ray absorptiometry was used to measure bone mineral density of rats,the values were compared with preoperative to ensure the model successfully,and the osteoporosis rats' BMSCs were cultured by bone marrow adherent cultured and the BMSCs morphology was observed under a phase contrast microscope upside down. The osteoporosis rats' BMSCs at the 2nd generation(p2)were randomly divided into experimental and control groups and were added complete medium(containing rhBMP-2)and osteogenic induced liquid,respectively. Two weeks later,the formation of cell calcium nodules were detected by Alizarin red staining method,alkaline phosphatase activity was measured by enzyme standard instrument and the expression of VEGF was detected by RT-PCT method. Results:(1)Whole body bone mineral density of rats before and after operation were(0.179 ± 0.007),(0.158 ± 0.006)g/cm2,there was statistically significant(t=4.180,P<0.05).(2)Alizarin red staining at 2 weeks after osteogenesis induced by BMSCs(P2)in the experimental group had more strong dyeing effect than the control group obviously.(3)Alkaline phosphatase activity at 2 weeks after osteogenesis induced by BMSCs(P2)of the experimental group(15.62 ± 1.27)ug/gprot was significantly higher than that of the control group(8.62±0.93)ug/gprot,there was statistically significant(t=7.709,P<0.01).(4)The expression of VEGF at 2 weeks af?ter osteogenesis induced by BMSCs(P2)of the experimental group 3.723±0.143 was significantly higher than that of the con?trol group 0.950±0.072,there was statistically significant(t=29.462,P<0.01). Conclusion:RhBMP-2 can improve the in vi? tro osteogenesis ability of ovary osteoporosis rat BMSCs,promote the VEGF expression of osteogenesis factor. Regulating the VEGF expression may be one of the mechanisms of BMP-2 to participate in bone metabolism.
