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| Effects of eclipta extract on migration and osteogenic differentiation of mouse bone marrow-derived mesenchymal stem cells |
| Hits 590 Download times 502 Received:October 27, 2021 |
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| DOI
10.11656/j.issn.1672-1519.2022.02.26 |
| Key Words
eclipta;bone marrow mesenchymal stem cell;cell migration;CXCR4;osteogenic differentiation |
| Author Name | Affiliation | E-mail | | AN Ran | Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | CAI Mingqi | Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | QIN Xiaoyan | Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | WEI Qiu | Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | YANG Yun | Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | MAO Haoping | Key Laboratory of Pharmacology of Traditional Chinese Medical Formulae, Ministry of Education, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | haoping_mao@126.com |
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| Abstract
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| [Objective] To investigate the effects of ecliptaherba (EHE) on the migration and osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) from mice.[Methods] BMMSCs were extracted from the femur and tibia of the hindlimb of 6-week C57BL/6J female mice, and the expression of cell surface molecules was detected by flow cytometry. ALP staining, alizarin red staining, toluidine blue staining and oil red O staining were used to observe the multi-directional differentiation of BMMSCs into osteoblasts, adipocytes and chondrocytes. Transwell experiment was used to explore the effect of EHE on the migration of BMMSCs. Western blot was used to detect the effect of EHE on the expression of CXC chemokine receptor 4(CXCR4) in BMMSCs. The effect of EHE on osteogenic induction of BMMSCs was observed by ALP activity detection, ALP staining and alizarin red staining.[Results] After the extraction and separation of mouse primary BMMSCs, the cells adhered to the wall and grew in a long spindle shape. The results of flow cytometry showed that the surface molecular expression rates of BMMSCs were as follows:Sca-1 was 86%, CD29 was 99%, cd140 α 88.7%, 63.7% for CD90, 14% for CD45 and 1% for CD34. Induced by osteoblast inducer, black stained osteoblasts can be seen after ALP staining, and red mineralized nodular osteoblasts can be seen after alizarin red staining;induced by chondrocyte inducer, blue stained chondrocytes were observed after toluidine blue staining;after being induced by adipocyte inducer, orange fat drop like adipocytes were observed after oil red O staining. The results of Transwell experiment showed that compared with the control group, 50 ng/mL stromal cell derived factor (SDF-1) and 0.1 μg/mL and 1 μg/mL EHE could significantly increase the number of permeable membranes of BMMSCs (P<0.05). The results of Western blot showed that compared with the control group, 0.1 μg/mL EHE could promote the expression of CXCR4 protein (P<0.05). ALP activity test, ALP staining and alizarin red staining showed that compared with the control group, osteogenic inducer significantly induced BMMSCs to differentiate into osteoblasts (P<0.05);compared with osteogenic inducer group, 1μg/mL EHE could significantly induce BMMSCs to differentiate into osteoblasts (P<0.05).[Conclusion] EHE can promote the migration and osteogenic differentiation of BMMSCs, and its mechanism of regulating the migration of BMMSCs may be related to promoting the expression of CXCR4 protein. |
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