目的 探讨血管内皮祖细胞(endothelial progenitor cell,EPC)对骨质疏松(OP)大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMSC)自主脂肪化的影响。方法 选6周龄雌性 SD 大鼠,建立OP模型。体外分离、培养大鼠EPC、BMSC。分别取第3、6代BMSC及原代EPC,建立EPC/BMSC 间接共培养体系。实验分组:假手术组BMSC(BMSC sham)、OP组BMSC(BMSC ovx)、OP BMSC /EPC共培养组(BMSC ovx/EPC);MTT法检测细胞增殖情况;油红 O染色检测成脂分化情况,real- time PCR检测成脂分化标志物PPARγ2的mRNA表达水平。结果 BMSC ovx/EPC 组BMSC的增殖能力显著高于BMSC ovx组;BMSC ovx/EPC组自主脂肪细胞分化水平明显低于BMSC ovx组。结论 EPC可通过间接接触提高骨质疏松大鼠BMSC的增殖能力,并能有效抑制OP大鼠BMSC的自主脂肪化。
Abstract
Objective To investigate the effect of endothelial progenitor cells (EPCs) on adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) of osteoporosis rats in order to provide a new strategy for prevention and treatment of osteoporosis.Methods An osteoporosis animal model was established with six-week-old female SD rats. EPCs and BMSCs of these rats were cultivated and identified in vitro. BMSCs at passage 3 and 6 were used respectively to establish a BMSC-EPC indirect co-culture system on transwell inserts. The experimental groups included the BMSC sham group, BMSC ovx group and BMSC ovx/EPC indirect co-culture group. MTT assay was applied to measure the cellular proliferation while Oil Red O solution was used to detect lipid droplets within the cells. Real-time PCR analyses were performed to analyze the expression of adipogenic marker PPARγ2.Results Under indirect co-culture conditions, EPCs could significantly promote the proliferation of BMSCs(ovx) and greatly inhibit the cells’capacity of self-adipogenic differentiation. Co-cultured EPCs resulted in a better proliferative ability of BMSCs in the BMSC ovx/EPC group than in the BMSC ovx group. Furthermore, BMSCs in the BMSC ovx/EPC group had a lower self-adipogenic differentiation potential than in the BMSC ovx group, which was evidenced by Oil Red O staining quantification and mRNA expression levels of adipogenic marker gene PPARg2.Conclusions Under indirect co-culture conditions, EPCs can enhance the stem cells’ proliferation but notably inhibit autonomic adipogenic differentiation of BMSCs of osteoporosis rats.
关键词
血管内皮祖细胞 /
间充质干细胞 /
骨质疏松 /
细胞增殖 /
成脂分化
Key words
endothelial progenitor cell /
mesenchymal stem cell /
osteoporosis /
cell proliferation /
adipogenic differentiation
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基金
国家自然科学基金资助项目(81602393)
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