去卵巢骨质疏松大鼠骨髓间充质干细胞成骨分化能力研究

目的　探讨去卵巢骨质疏松大鼠骨髓间充质干细胞(MSCs)成骨分化的特点。方法　选用3月龄健康SD雌性大鼠行双侧卵巢切除术,建立骨质疏松症的动物模型。实验分为正常大鼠髓间充质干细胞组(MSCs controlgroup )、骨质疏松大鼠骨髓间充质干细胞组(MSCs ovx group)、正常骨髓间充质干细胞成骨诱导组(OSI controlgroup)、骨质疏松骨髓间充质干细胞成骨诱导组(OSI ovx group)。使用密度梯度离心法分别获取正常大鼠和骨质疏松大鼠MSCs,体外培养传至3、4代后,流式细胞技术检测MSCs control group和MSCs ovx group细胞周期及增殖指数(PI) ;加入含有地塞米松(10 - 8m ol/L) ,β-甘油磷酸钠(10 - 2 mol/L) ,抗坏血酸(5 0 μg/m l)的成骨诱导液进行成骨诱导,采用磷酸对硝基苯测定方法检测碱性磷酸酶(AL P)表达量,同位素标记方法检测骨钙素(BGP)分泌量。结果　1PI:MSCs control group高于MSCs ovx group(P<0 .0 5 )。2 AL P的表达量:成骨诱导第7d和14 d时,OSI各组均明显高于相应的MSCs组(P<0 .0 5 ) ;OSI control组明显高于OSI ovx组(P<0 .0 5 )。随着时间延长,OSI各组AL P表达量呈升高趋势。3BGP的分泌量:成骨诱导14 d、2 1d、2 8d时,OSI各组均明显高于相应的MSCs组(P<0 .0 5 ) ;OSI control组明显高于OSI ovx组(P<0

Osteogenic Potential of Bone Marrow Mesenchymal Stem Cells from Ovariectomied Osteoporotic Rat

OBJECTIVE: To investigate the difference of osteogenic potential of bone marrow mesenchymal stem cells (MSCs) between healthy rats and osteoporotic rats. METHODS: We established the animal model of osteoporosis by performing ovariectom on the 3-month-old female Sprague-Dawley rats. Bone marrow mesenchymal stem cells(MSCs) were isolated from the rats of control group and of ovariectomized (ovx) group by means of the density-gradient centrifugation method, and the 3rd-4th passage MSCs were used in all the experiments. The experiments comprised 4 groups: (1) Marrow mesenchymal stem cells control group (MSCs control group); (2) Marrow mesenchymal stem cells ovx group (MSCs ovx group); (3) Osteogenesis induction control group (OSI control group); (4) Osteogenesis induction ovx group (OSI ovx group). Cell cycle and proliferation index (PI) of MSCs were detected by flow cytometry. The expression of alkaline phosphatase (ALP) was detected by dynamics method with substrate of phosphoric acid para-Nitro benzene. The levels of osteocalcin were detected with the isotope labelling method. RESULTS: (1) PI of MSCs was lower in MSCs ovx group than in MSCs control group. (2) The expression of alkaline phosphatase (ALP) was much higher in OSI control group than in the MSCs control group; the expression of alkaline phosphatase (ALP) was much higher in the OSI control group than in OSI ovx group after 7-day and 14-day osteogenic induction. (3) The level of osteocalcin was much higher in the OSI control group than in the MSCs control group after 14-day, 21-day, 28-day osteogenic induction. The level of osteocalcin was much higher in the OSI control group than in the OSI ovx group. CONCLUSION: Both the proliferative potential and the osteogenic potential of bone marrow mesenchymal stem cells (MSCs) from the ovariectomized osteoporotic rat are decreased.