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| 基于Hippo信号通路探讨鹿茸提取液对BMSCs成骨分化的影响 |
| To investigate the effect of pilose antler extract on BMSCs osteogenic differentiation based on Hippo signaling pathway |
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| DOI：10.3969/j.issn.1006.7108.2022.10.017 |
| 中文关键词: 鹿茸提取液 成骨分化 Hippo信号通路 骨髓间充质干细胞 |
| 英文关键词:pilose antler extract osteogenic differentiation hippo signaling pathway bone marrow mesenchymal stem cells |
| 基金项目:国家重点研发计划（2018YFC1704301）；国家自然科学基金青年科学基金项目（82104709）；辽宁省自然科学基金面上项目（2020-MS-222）；2016年百千万人才工程[2017(59)]；沈阳市中青年科技创新人才支持计划（RC210129）；辽宁中医药大学自然科学重点项目（2021LZY035） |
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| 中文摘要: |
| 目的 观察鹿茸提取液对SD大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）向成骨分化及Hippo信号通路的影响。方法 将3、4代BMSCs随机分为正常组、诱导组以及鹿茸组。酶联免疫吸附测定（enzyme linked immunosorbent assay，ELISA）法分别检测6、12 d以及18 d碱性磷酸酶(alkaline phosphatase，ALP)的活性及水平；茜素红染色法检测3组BMSCs钙结节生长增殖情况；qRT-PCR法和蛋白质印迹法（Western blot）检测RUNX2及Hippo信号通路 MST1和YAP的转录表达与蛋白表达水平。结果 ELISA实验结果表明，鹿茸提取液可以提高ALP的活性，12 d鹿茸组ALP的活性达到最高，18 d鹿茸组ALP的活性显著上调（P＜0.01）；茜素红实验结果显示，鹿茸组观察到大量且密集的钙结节形成，提示鹿茸提取液可有效促进骨髓间充质干细胞向成骨细胞转化；qRT-PCR的实验结果表明，与正常组相比，鹿茸组YAP和RUNX2 mRNA显著上调（P＜0.01）；Western blot法检测结果显示，鹿茸提取液可显著上调YAP蛋白表达水平（P＜0.01），并且明显上调RUNX2蛋白表达水平（P＜0.05）。结论 鹿茸提取液能够影响RUNX2以及Hippo信号通路中 MST1和YAP相关蛋白进而推进骨髓间充质干细胞向成骨细胞的转化。 |
| 英文摘要: |
| Objective To observe the effects of pilose antler extract on Bone marrow mesenchymal stem cells (BMSCs) differentiation and Hippo signaling pathway in SD rats. Methods BMSCs of the third and fourth generations were randomly divided into normal group, induced group and deer antler group. Enzyme Linked immunosorbent assay (ELISA) was used to determine the activity and content of alkaline phosphatase (ALP) at 6, 12 and 18 days, respectively. Alizarin red staining was used to detect the growth and proliferation of bone marrow mesenchymal stem cells in the three groups. The transcription and protein expression levels of MST1 and YAP in RUNX2 and Hippo signaling pathways were detected by RT-PCR and Western blot. Results The results of ELISA showed that the serum ALP activity was increased by antler antler extract. The serum ALP activity was the highest in antler antler group at 12 days, and significantly up-regulated in antler antler group at 18 days（P＜0.01）. Alizarin red assay showed that a large number of calcium nodules were observed in the pilose antler group, suggesting that pilose antler extract can effectively promote the transformation of bone marrow mesenchymal stem cells into osteoblasts. RT-PCR results showed that the mRNA of YAP and RUNX2 were significantly up-regulated in the antler group compared with the normal group（P＜0.01）. Western blot analysis showed that the expression of YAP protein and RUNX2 protein was significantly up-regulated by antler antler extract（P＜0.01）. Conclusion Pilose antler extract can affect MST1 and YAP related proteins in RUNX2 and Hippo signaling pathway and promote the transformation of bone marrow mesenchymal stem cells into osteoblasts. |
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