中国骨质疏松杂志
中國骨質疏鬆雜誌
중국골질소송잡지
CHINESE JOURNAL OF OSTEOPOROSIS
2014年
4期
382-386,403
,共6页
1α,25(OH)2D3%成骨细胞%RANKL%OPG
1α,25(OH)2D3%成骨細胞%RANKL%OPG
1α,25(OH)2D3%성골세포%RANKL%OPG
1α,25-( OH) 2 D3%Osteoblasts%RANKL%OPG
目的：研究高剂量1α，25（OH）2D3对RANKL/OPG表达的调控是否随成骨细胞（osteoblasts，OB）分化成熟而变化。方法体外诱导MC3T3-E1细胞成骨分化，并分别于成骨诱导第0、7、14、21、28 d向细胞加以1α，25（OH）2D3（10 nM）的刺激，4 h后提取细胞总RNA，进行RT-PCR检测OB RANKL及OPG mRNA表达水平。对照组则以无水乙醇刺激。同时，对不同分化阶段OB进行茜素红染色，以检测钙结节的形成。结果在OB分化过程中，对照组RANKL表达无显著变化；OPG 基因表达逐渐增多，在第14天达最高水平，之后逐渐下降；RANKL/OPG比值始终低于未成骨诱导时的RANKL/OPG比值。1α，25（ OH）2 D3持续促进RANKL基因的表达，并且持续抑制OPG基因表达；实验组RANKL/OPG比值一直高于对照组，尤其在矿化阶段，差异有统计学意义。结论1α，25（OH）2D3对RANKL和OPG的调控随OB分化成熟而变化。
目的：研究高劑量1α，25（OH）2D3對RANKL/OPG錶達的調控是否隨成骨細胞（osteoblasts，OB）分化成熟而變化。方法體外誘導MC3T3-E1細胞成骨分化，併分彆于成骨誘導第0、7、14、21、28 d嚮細胞加以1α，25（OH）2D3（10 nM）的刺激，4 h後提取細胞總RNA，進行RT-PCR檢測OB RANKL及OPG mRNA錶達水平。對照組則以無水乙醇刺激。同時，對不同分化階段OB進行茜素紅染色，以檢測鈣結節的形成。結果在OB分化過程中，對照組RANKL錶達無顯著變化；OPG 基因錶達逐漸增多，在第14天達最高水平，之後逐漸下降；RANKL/OPG比值始終低于未成骨誘導時的RANKL/OPG比值。1α，25（ OH）2 D3持續促進RANKL基因的錶達，併且持續抑製OPG基因錶達；實驗組RANKL/OPG比值一直高于對照組，尤其在礦化階段，差異有統計學意義。結論1α，25（OH）2D3對RANKL和OPG的調控隨OB分化成熟而變化。
목적：연구고제량1α，25（OH）2D3대RANKL/OPG표체적조공시부수성골세포（osteoblasts，OB）분화성숙이변화。방법체외유도MC3T3-E1세포성골분화，병분별우성골유도제0、7、14、21、28 d향세포가이1α，25（OH）2D3（10 nM）적자격，4 h후제취세포총RNA，진행RT-PCR검측OB RANKL급OPG mRNA표체수평。대조조칙이무수을순자격。동시，대불동분화계단OB진행천소홍염색，이검측개결절적형성。결과재OB분화과정중，대조조RANKL표체무현저변화；OPG 기인표체축점증다，재제14천체최고수평，지후축점하강；RANKL/OPG비치시종저우미성골유도시적RANKL/OPG비치。1α，25（ OH）2 D3지속촉진RANKL기인적표체，병차지속억제OPG기인표체；실험조RANKL/OPG비치일직고우대조조，우기재광화계단，차이유통계학의의。결론1α，25（OH）2D3대RANKL화OPG적조공수OB분화성숙이변화。
Objective To investigate the effect of high-dose 1α,25-(OH)2D3 on the changes of RANKL and OPG mRNA expression during osteoblast ( OB) differentiation.Methods The osteogenic differentiation of MC3T3-E1 cells was induced in vitro.A 10nM concentration of 1α,25-(OH)2D3 was additioned into the medium at the 0, 7th, 14th, 21st, and the 28th day, respectively, in order to stimulate the osteogenic induction.Four hours after the stimulation, total RNA was extracted.And the expression of RANKL and OPG mRNA in OBs was detected using RT-PCR.Cells in control group were stimulated with absolute ethyl alcohol instead of 1α,25-( OH) 2 D3 .Meanwhile, Alizarin red staining was carried out at the indicated different time-point, in order to detect the formation of mineralized nodules.Results The expression of RANKL mRNA in control groups showed no significant change, while the expression of OPG mRNA increased and reached the peak level at the 14 th day, then decreased gradually.RANKL/OPG ratio was constanly lower than that at the base line (undifferentiated).1α,25-(OH)2D3 upregulated the expression of RANKL continuously, but it inhibited OPG gene expression.The RANKL/OPG ratio in study group was constantly higher than that in control group, especially during the mineralizing phase.And the difference was statistically significant ( P<0.05).Conclusion The regulating effect of 1α,25-( OH)2D3 on the expression of RANKL and OPG changes at various differential stage of OBs.
