中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2015年
4期
249-253,封3
,共6页
高飞%葛志敏%郭中豪%杨静
高飛%葛誌敏%郭中豪%楊靜
고비%갈지민%곽중호%양정
甲状旁腺激素肽1-34%间质干细胞%骨髓%成脂分化%成骨分化
甲狀徬腺激素肽1-34%間質榦細胞%骨髓%成脂分化%成骨分化
갑상방선격소태1-34%간질간세포%골수%성지분화%성골분화
Teriparatide%Mesenchymal stem cells%Bone marrow%Adipogenic differentiation%Osteogenic differentiation
目的 观察甲状旁腺激素肽1-34(PTH1-34)对大鼠骨髓间充质干细胞(BMSCs)成脂分化的影响.方法 ①应用贴壁培养法分离大鼠BMSCs,观察细胞的形态学特征,流式细胞仪检测细胞表面标志物.②诱导BMSCs定向分化为成骨细胞和脂肪细胞,并通过组织染色的方法进行鉴定.③以P3代BMSCs为体外试验模型,不同浓度PTH 1-34(0、10-10、10-9、10-8 mol/L)分别作用于加入成脂诱导液的BMSCs,14d后采用ELISA法测定脂蛋白脂肪酶(LPL)活性,采用反转录(RT)-PCR法测定ALP和过氧化物酶体增殖物激活受体(PPARγ)-2 mRNA的表达.采用方差分析和LSD-t检验进行统计学分析.结果 ①P3代细胞表达抗原CD29、CD44,不表达抗原CD45,符合BMSCs表面标志物特征.②成脂诱导液诱导BMSCs分化21d后,油红0染色可见胞内脂滴出现,成骨诱导28 d后,茜素红染色可见红色钙结节.③与空白对照组比较,不同浓度PTH 1-34(10-10、10-9、10-8 mol/L)组LPL的表达量分别为11.20±0.16、7.62±0.48、5.84±0.57、5.32±0.52,PPARγ-2 mRNA的表达量分别是2.80±0.05、1.36±0.23、0.94±0.11、0.78±0.04,ALP活性分别为0.191±0.016、0.333±0.024、0.549±0.025、0.684±0.021,随着PTH1-34浓度的增加,IPL的表达量及PPARγ-2 mRNA的表达量下降,ALP活性增加,并呈剂量依赖性(P＜0.05).结论 PTH1-34抑制BMSCs成脂分化,促进其成骨分化,并呈剂量依赖性.
目的 觀察甲狀徬腺激素肽1-34(PTH1-34)對大鼠骨髓間充質榦細胞(BMSCs)成脂分化的影響.方法 ①應用貼壁培養法分離大鼠BMSCs,觀察細胞的形態學特徵,流式細胞儀檢測細胞錶麵標誌物.②誘導BMSCs定嚮分化為成骨細胞和脂肪細胞,併通過組織染色的方法進行鑒定.③以P3代BMSCs為體外試驗模型,不同濃度PTH 1-34(0、10-10、10-9、10-8 mol/L)分彆作用于加入成脂誘導液的BMSCs,14d後採用ELISA法測定脂蛋白脂肪酶(LPL)活性,採用反轉錄(RT)-PCR法測定ALP和過氧化物酶體增殖物激活受體(PPARγ)-2 mRNA的錶達.採用方差分析和LSD-t檢驗進行統計學分析.結果 ①P3代細胞錶達抗原CD29、CD44,不錶達抗原CD45,符閤BMSCs錶麵標誌物特徵.②成脂誘導液誘導BMSCs分化21d後,油紅0染色可見胞內脂滴齣現,成骨誘導28 d後,茜素紅染色可見紅色鈣結節.③與空白對照組比較,不同濃度PTH 1-34(10-10、10-9、10-8 mol/L)組LPL的錶達量分彆為11.20±0.16、7.62±0.48、5.84±0.57、5.32±0.52,PPARγ-2 mRNA的錶達量分彆是2.80±0.05、1.36±0.23、0.94±0.11、0.78±0.04,ALP活性分彆為0.191±0.016、0.333±0.024、0.549±0.025、0.684±0.021,隨著PTH1-34濃度的增加,IPL的錶達量及PPARγ-2 mRNA的錶達量下降,ALP活性增加,併呈劑量依賴性(P＜0.05).結論 PTH1-34抑製BMSCs成脂分化,促進其成骨分化,併呈劑量依賴性.
목적 관찰갑상방선격소태1-34(PTH1-34)대대서골수간충질간세포(BMSCs)성지분화적영향.방법 ①응용첩벽배양법분리대서BMSCs,관찰세포적형태학특정,류식세포의검측세포표면표지물.②유도BMSCs정향분화위성골세포화지방세포,병통과조직염색적방법진행감정.③이P3대BMSCs위체외시험모형,불동농도PTH 1-34(0、10-10、10-9、10-8 mol/L)분별작용우가입성지유도액적BMSCs,14d후채용ELISA법측정지단백지방매(LPL)활성,채용반전록(RT)-PCR법측정ALP화과양화물매체증식물격활수체(PPARγ)-2 mRNA적표체.채용방차분석화LSD-t검험진행통계학분석.결과 ①P3대세포표체항원CD29、CD44,불표체항원CD45,부합BMSCs표면표지물특정.②성지유도액유도BMSCs분화21d후,유홍0염색가견포내지적출현,성골유도28 d후,천소홍염색가견홍색개결절.③여공백대조조비교,불동농도PTH 1-34(10-10、10-9、10-8 mol/L)조LPL적표체량분별위11.20±0.16、7.62±0.48、5.84±0.57、5.32±0.52,PPARγ-2 mRNA적표체량분별시2.80±0.05、1.36±0.23、0.94±0.11、0.78±0.04,ALP활성분별위0.191±0.016、0.333±0.024、0.549±0.025、0.684±0.021,수착PTH1-34농도적증가,IPL적표체량급PPARγ-2 mRNA적표체량하강,ALP활성증가,병정제량의뢰성(P＜0.05).결론 PTH1-34억제BMSCs성지분화,촉진기성골분화,병정제량의뢰성.
Objective To observe the effect of different concentrations of parathyroid hormone 1-34 on the adipogenic potential of rat bone marrow mesenchymal stem cells (BMSCs).Methods ① Rat bone marrow mesenchymal cells were separated and expanded by adherent culture.The morphology of cells was observed and cell surface markers were examined by flow cytometry.② The multi-lineage differentiation capability of cells was examined by culturing cells under conditions favorable for adipogenic and osteogenic differentiation.③ Taken P3 of BMSCs for test,different concentrations of PTH1-34 (0,10-10,10-9,10-8 mol/L) were used to stimulate BMSCs respectively,14 days later,lipoprotein lipase (LPL) activity were measured by enzyme linked immuno-sorbent assay (ELISA),mRNA expression of alkaline phosphatase (ALP) and PPARγ-2 were measured by realiime polymerase chain reaction (PCR).④ Statistical analysis:data were presented as x±s.All statistical analysis was performed with windows Statistical Praduct and Serice Solutions (SPSS) 13.0.One-way analysis of variance (ANOVA) was applied to determine the difference between groups.Least signisicant difference (LSD) was used to determine the difference between the two randomized groups.Differences were considered significant at a value of P＜0.05.Results The cells expressed CD44,CD29 but without expression of CD45.By culturing in adipogenic medium for 3 weeks and in osteogenic medium for 4 weeks respectively,and then identified by oil red O and Alizarin red,the cells were successfully induced to adipocytes and osteogenesis.Expressions of LPL were 11.20±0.16,7.62±0.48,5.84±0.57,5.32±0.52,mRNA expressions of PPARγ-2 were 2.80±0.05,1.36±0.23,0.94-±0.11,0.78±0.04,ALP activity were 0.191 ±0.016,0.333±0.024,0.549±0.025,0.684±0.021 respectively.Compared with the control group,different concentrations of PTH1-34 groups could decrease mRNA expression of LPL and PPARγ-2.ALP activity were increased(P＜0.05).Conclusion PTH1-34 inhibits BMSCs of adipogenic differentiation and promotes osteogenic differentiation in a dose-dependent manner.
