CAJ | 학술논문

目的:探讨成骨细胞中破骨细胞分化抑制因子(OCIL)基因敲减后护骨素(OPG)和核因子κB受体活化因子配基(RANKL)mRNA的表达水平及其对破骨细胞分化的影响.方法:构建OCIL特异性shRNA表达载体并转染至细胞,分为pRNAT-N1组、pRNAT-N2组、pRNAT-N3组和阴性对照转染组.考察与阴性对照转染组比较,各组对OCIL mRNA水平的抑制率.将对OCIL mRNA干扰效果最佳的shRNA表达载体(pRNAT-N1组)和对照载体pRNAT-U6.1/Neo/CTL(control组)分别瞬时转染UMR106细胞,考察2组RANKL和OPG表达水平.各转染组条件培养基干预骨髓细胞后,观察PBS+control shRNA(vehicle组)、PTH+control shRNA(PTH组)及PTH+ OCILshRNA(shRNA组)破骨细胞样细胞(OLC)数量.结果:(1)与阴性对照转染组比较,pRNAT-N1、pRNAT-N2、pRNAT-N3对OCIL mRNA水平抑制率分别为64％、28％和59％.(2)pRNAT-N1转染UMR106成骨细胞后,RANKL mRNA表达水平明显升高,而OPG mRNA表达水平明显降低,转染后24 h RANKL:OPG比值较control组升高5.1倍.(3)vehicle组、PTH组及shRNA组OLC数分别为(1.83±0.75)个/孔、(49.67±6.77)个/孔及(65.50±7.34)个/孔,差异有统计学意义(P＜0.05).结论:RANKL/OPG系统参与了OCIL对破骨细胞分化的调节作用.
목적:탐토성골세포중파골세포분화억제인자(OCIL)기인고감후호골소(OPG)화핵인자κB수체활화인자배기(RANKL)mRNA적표체수평급기대파골세포분화적영향.방법:구건OCIL특이성shRNA표체재체병전염지세포,분위pRNAT-N1조、pRNAT-N2조、pRNAT-N3조화음성대조전염조.고찰여음성대조전염조비교,각조대OCIL mRNA수평적억제솔.장대OCIL mRNA간우효과최가적shRNA표체재체(pRNAT-N1조)화대조재체pRNAT-U6.1/Neo/CTL(control조)분별순시전염UMR106세포,고찰2조RANKL화OPG표체수평.각전염조조건배양기간예골수세포후,관찰PBS+control shRNA(vehicle조)、PTH+control shRNA(PTH조)급PTH+ OCILshRNA(shRNA조)파골세포양세포(OLC)수량.결과:(1)여음성대조전염조비교,pRNAT-N1、pRNAT-N2、pRNAT-N3대OCIL mRNA수평억제솔분별위64％、28％화59％.(2)pRNAT-N1전염UMR106성골세포후,RANKL mRNA표체수평명현승고,이OPG mRNA표체수평명현강저,전염후24 h RANKL:OPG비치교control조승고5.1배.(3)vehicle조、PTH조급shRNA조OLC수분별위(1.83±0.75)개/공、(49.67±6.77)개/공급(65.50±7.34)개/공,차이유통계학의의(P＜0.05).결론:RANKL/OPG계통삼여료OCIL대파골세포분화적조절작용.