CAJ | 학술논문

目的探讨系统性红斑狼疮(SLE)患者骨髓间充质干细胞(MSCs)多向分化能力是否存在异常. 方法密度梯度离心法和贴壁筛选法分离培养MSCs,定向诱导骨髓MSCs向脂肪和成骨细胞分化,脂肪细胞经油红O染色鉴定并定量,成骨细胞经茜素红S染色鉴定.将骨髓MSCs与羟基磷灰石共孵育,将共孵育物植于裸鼠皮下,8周后常规苏木素-伊红(HE)染色.反转录-聚合酶链反应(RT-PCR)检测过氧化物酶体增殖物激活受体γ-2(PPARγ-2)、脂蛋白酶(LPL)、Runx2/CBFA1、降钙素(osteocalcin) mRNA的表达水平.结果 SLE骨髓MSCs分化成脂肪细胞比例低于健康埘照组,油红O染色定量低于健康对照组[(35±7)%与(80±5)%],(0.14±0.04与0.27±0.04),LPL mRNA表达低于健康对照组(0.369±0.020与0.481±0.038),两组PPARγ-2 mRNA表达差异无统计学意义(0.421±0.052与0.441±0.012).SLE骨髓MSCs分化成骨细胞后形成钙结节低于健康对照组[(35±4)%与(45±4)%],Runx2/CBFA1、osteocalcin mRNA表达低于健康对照组(0.371±0.000与0.563±0.069),(0.819±0.023与0.962±0.049);羟基磷灰石与SLE患者骨髓MSCs共孵育后移植裸鼠皮下8周后,成骨细胞形成明显少于健康埘照组. 结论 SLE骨髓MSCs成脂和成骨分化能力存在异常,提示SLE患者骨髓MSCs存在缺陷.
목적 탐토계통성홍반랑창(SLE)환자골수간충질간세포(MSCs)다향분화능력시부존재이상. 방법 밀도제도리심법화첩벽사선법분리배양MSCs,정향유도골수MSCs향지방화성골세포분화,지방세포경유홍O염색감정병정량,성골세포경천소홍S염색감정.장골수MSCs여간기린회석공부육,장공부육물식우라서피하,8주후상규소목소-이홍(HE)염색.반전록-취합매련반응(RT-PCR)검측과양화물매체증식물격활수체γ-2(PPARγ-2)、지단백매(LPL)、Runx2/CBFA1、강개소(osteocalcin) mRNA적표체수평.결과 SLE골수MSCs분화성지방세포비례저우건강시조조,유홍O염색정량저우건강대조조[(35±7)%여(80±5)%],(0.14±0.04여0.27±0.04),LPL mRNA표체저우건강대조조(0.369±0.020여0.481±0.038),량조PPARγ-2 mRNA표체차이무통계학의의(0.421±0.052여0.441±0.012).SLE골수MSCs분화성골세포후형성개결절저우건강대조조[(35±4)%여(45±4)%],Runx2/CBFA1、osteocalcin mRNA표체저우건강대조조(0.371±0.000여0.563±0.069),(0.819±0.023여0.962±0.049);간기린회석여SLE환자골수MSCs공부육후이식라서피하8주후,성골세포형성명현소우건강시조조. 결론 SLE골수MSCs성지화성골분화능력존재이상,제시SLE환자골수MSCs존재결함.
Objective To investigate the muhilineage differentiation potential of bone marrow-derived mesenchymM stem eels (MSCs) in patients with systemic lupus erythematosus (SLE).Methods Density gradient centrifugation and plastic adherence methods were used for isolation of marrow-derived MSCs.Then tIIeir differentiation potentiality to lipoblasts and osteoblasts waft tested.MSCs loading on hydroxyapatite were elnbedded in the nude mouse's subcutaneous tissues.Eight weeks later.osteogenesis was evaluated by HE staining.PPA Rγ2,LPL,Runx2/CBFA1,osteocalcin gene expression in MSCs after differentiation were examined by RT-PCR.Results The positive rates of lipoblasts stained by oil red O and optical density in SLE were decreased than in the control group[(35±7)% vs (80±5)%] (0.14±0.04 vs 0.27±0.04),and the positive rates of osteoblasts stained by Alizarin Red S in SLE were decreased than those in the control group [(35±4)% vs (45±4)%].Osteoblast differentiation in the SLE group was less than that of the contro]group.The mRNA expression of LPL (0.369±0.020 vs 0.481±0.038).Runx2/CBFA1 (0.371±0.000 vs 0.563±0.069).osteoealcin (0.819±0.023 vs 0.962±0.049) of MSCs after difierentiation in the SLE group was decreased than that of the control group.There was no significant difference in the expression of PPARγ2 mRNA between SLE and controI group (0.421±0.052 vs 0.441±0.012).Conelusion MSCs from SLE have abnormalities in osteogenie and adipogenic differentiation potential.