Journal of Prevention and Treatment for Stomatological Diseases ›› 2021, Vol. 29 ›› Issue (7): 449-455.DOI: 10.12016/j.issn.2096-1456.2021.07.003
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ZUO Xinhui1(),LI Jun1,HAN Xiangzhen1,LIU Xiaoyuan1,HE Huiyu1,2(
)
Received:
2020-08-18
Revised:
2021-01-05
Online:
2021-07-20
Published:
2021-04-19
Contact:
Huiyu HE
Supported by:
通讯作者:
何惠宇
作者简介:
左新慧,住院医师,硕士研究生,Email: 基金资助:
CLC Number:
ZUO Xinhui,LI Jun,HAN Xiangzhen,LIU Xiaoyuan,HE Huiyu. Effects of hypoxia inducible factor-1α on osteogenic differentiation and angiogenesis related factors of bone marrow mesenchymal stem cells[J]. Journal of Prevention and Treatment for Stomatological Diseases, 2021, 29(7): 449-455.
左新慧,李君,韩祥祯,刘小元,何惠宇. 低氧诱导因子-1α对骨髓间充质干细胞成骨分化与血管生成相关因子的影响[J]. 口腔疾病防治, 2021, 29(7): 449-455.
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Primers | Sequence |
---|---|
GAPDH | F:5’-GGCACAGTCAAGGCTGAGAATG-3’ R:5’-ATGGTGGTGAAGACGCCAGTA-3’ |
HIF-1α | F:5’-CACCGCAACTGCCACCACTG-3’ R:5’-TGAGGCTGTCCGACTGTGAGTAC-3’ |
Runx2 | F:5’-CCATAACGGTCTTCACAAATCCT-3’ R:5’-TCTGTCTGTGCCTTCTTGGTTC-3’ |
PDGF-BB | F:5’-TCTCTGCTGCTACCTGCGTCTG-3’ R:5’-AAGGAGCGGATGGAGTGGTCAC-3’ |
TGF-β | F:5’-GGCACCATCCATGACATGAACCG-3’ R:5’-GCCGTACACAGCAGTTCTTCTCTG-3’ |
Table 1 Synthetic primers used for RT-PCR
Primers | Sequence |
---|---|
GAPDH | F:5’-GGCACAGTCAAGGCTGAGAATG-3’ R:5’-ATGGTGGTGAAGACGCCAGTA-3’ |
HIF-1α | F:5’-CACCGCAACTGCCACCACTG-3’ R:5’-TGAGGCTGTCCGACTGTGAGTAC-3’ |
Runx2 | F:5’-CCATAACGGTCTTCACAAATCCT-3’ R:5’-TCTGTCTGTGCCTTCTTGGTTC-3’ |
PDGF-BB | F:5’-TCTCTGCTGCTACCTGCGTCTG-3’ R:5’-AAGGAGCGGATGGAGTGGTCAC-3’ |
TGF-β | F:5’-GGCACCATCCATGACATGAACCG-3’ R:5’-GCCGTACACAGCAGTTCTTCTCTG-3’ |
Figure 1 Cell culture of BMSCs(× 50) a: the primary cells were round; b: P3 cells, the cells adhered to the wall and grew in long fusiform or whirlpool shape;BMSCs: bone marrow mesenchymal stem cells
Figure 2 BMSC transfected with plasmid after 48 h (× 100) BMSCs that were successfully transfected with DNA plasmid showed green fluorescence expression, and the cell growth density was approximately 75%. The expression level of the low expression experimetal group was approximately 76%, and that of the control group was approximately 70%. The fluorescence level of the overexpression experimetal group was 73%, and that of the control group was approximately 68%. BMSCs: bone marrow mesenchymal stem cells
Figure 3 BMSC alizarin red staining results for each group(×50) The area of calcium nodules in the low expression group was smaller than that in the control group and smaller than the area of nodules in the overexpression experimental group. The area of red calcium nodules in the overexpression experimental group was significantly larger than that in the control group, and the calcification area in each group increased significantly with increasing osteogenic induction time. BMSCs: bone marrow mesenchymal stem cells
Gene names | Low expression experimental group | Low expression control group | t | P | Over expression experimental group | Over expression control group | t | P |
---|---|---|---|---|---|---|---|---|
HIF-1α | 0.373 ± 0.020 | 1.023 ± 0.231 | 22.522 | <0.001 | 1.563 ± 0.128 | 1.013 ± 0.171 | 0.896 | <0.001 |
Runx2 | 0.618 ± 0.012 | 1.025 ± 0.229 | 32.776 | <0.001 | 25.462 ± 0.406 | 1.023 ± 0.204 | 6.278 | <0.001 |
PDGF-BB | 0.097 ± 0.011 | 1.009 ± 0.155 | 16.531 | <0.001 | 3.322 ± 0.111 | 1.009 ± 0.147 | 1.705 | <0.001 |
TGF-β | 0.447 ± 0.013 | 1.023 ± 0.222 | 9.203 | <0.001 | 1.734 ± 0.016 | 1.006 ± 0.112 | 17.389 | <0.001 |
Table 2 mRNA expression of osteogenic differentiation and angiogenesis related factors $\bar{x}\pm s$
Gene names | Low expression experimental group | Low expression control group | t | P | Over expression experimental group | Over expression control group | t | P |
---|---|---|---|---|---|---|---|---|
HIF-1α | 0.373 ± 0.020 | 1.023 ± 0.231 | 22.522 | <0.001 | 1.563 ± 0.128 | 1.013 ± 0.171 | 0.896 | <0.001 |
Runx2 | 0.618 ± 0.012 | 1.025 ± 0.229 | 32.776 | <0.001 | 25.462 ± 0.406 | 1.023 ± 0.204 | 6.278 | <0.001 |
PDGF-BB | 0.097 ± 0.011 | 1.009 ± 0.155 | 16.531 | <0.001 | 3.322 ± 0.111 | 1.009 ± 0.147 | 1.705 | <0.001 |
TGF-β | 0.447 ± 0.013 | 1.023 ± 0.222 | 9.203 | <0.001 | 1.734 ± 0.016 | 1.006 ± 0.112 | 17.389 | <0.001 |
Figure 4 Protein expression of osteogenic differentiation and angiogenesis related factors 1: low expression experimental group; 2: low expression control group; 3: over expression experimental group; 4: over expression control group; HIF-1α: hypoxia inducible factor-1α; Runx2: runt-related transcription factor 2; PDGF-BB: platelet derived growth factor-BB; TGF-β: transforming growth factor-β; ***P<0.001
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