[1]蘇華斌,胡波涌,葉俊杰,等.miR-449對骨髓間充質干細胞成骨分化調控的機制研究[J].醫學信息,2020,(04):69-71,86.[doi:10.3969/j.issn.1006-1959.2020.04.021]
 SU Hua-bin,HU Bo-yong,YE Jun-jie,et al.The Mechanism of MiR-449 on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells[J].Medical Information,2020,(04):69-71,86.[doi:10.3969/j.issn.1006-1959.2020.04.021]
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miR-449對骨髓間充質干細胞成骨分化調控的機制研究()
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醫學信息[ISSN:1006-1959/CN:61-1278/R]

卷:
期數:
2020年04期
頁碼:
69-71,86
欄目:
論著
出版日期:
2020-02-15

文章信息/Info

Title:
The Mechanism of MiR-449 on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
文章編號:
1006-1959(2020)04-0069-04
作者:
蘇華斌胡波涌葉俊杰
(廣州市第八人民醫院外科1,骨科2,病理科3,廣東 廣州 510070)
Author(s):
SU Hua-binHU Bo-yongYE Jun-jieet al
(Department of Surgery1,Department of Orthopaedics2,Department of Pathology3,Eighth People’s Hospital of Guangzhou,Guangzhou 510070,Guangdong,China)
關鍵詞:
miR-449 a/b間充質干細胞成骨分化
Keywords:
miR-449 a/bMesenchymal stem cellsOsteogenic differentiation
分類號:
R329
DOI:
10.3969/j.issn.1006-1959.2020.04.021
文獻標志碼:
A
摘要:
目的 探究miR-449是否具有調控骨髓間充質干細胞(MSCs)成骨分化的作用。方法 分離、培養骨髓MSCs,采用流式細胞儀檢測CD44、CD29、CD34、CD45等表面相關標志抗原的表達鑒定骨髓MSCs;將MSCs分為對照組、miR-449a組、miR-449b組及anti-miR-449組,對照組用成骨誘導培養基培養,其余各組分別經miR-449a/b或inhibitors轉染后在成骨誘導培養基培養,分別于培養第3、6、9、12天檢測各組堿性磷酸酶活性,qRT-PCR檢測miR-449a/b對成骨特異性基因Runx2、Osterix表達的影響。結果 ①第4代間充質干細胞CD44、CD29表達陽性,而CD34 和CD45表達陰性,符合骨髓間充質干細胞的特征。②成骨誘導培養3、6、9、12 d后,MSCs細胞堿性磷酸酶活性較上一時間點升高,且呈時間依賴性,差異有統計學意義(P<0.05);MSCs成骨誘導培養3、6、9、12 d后,Runx2、Osterix表達水平逐漸升高。③miR-449a組和miR-449b組AKP活性低于對照組,而anti-miR-449組AKP活性升高,差異有統計學意義(P<0.05);miR-449a組和miR-449b組 Runx2、Osterix表達較對照組下調,差異有統計學意義(P<0.05)。結論 本次實驗成功分離、培養了骨髓MSCs,其具有體外成骨分化能力,而miR-449能抑制骨髓MSCs向成骨分化。
Abstract:
Objective To investigate whether miR-449 can regulate the osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs). Methods Bone marrow MSCs were isolated and cultured. Flow cytometry was used to detect the expression of surface-associated marker antigens such as CD44, CD29, CD34, and CD45. Bone marrow MSCs were identified. The MSCs were divided into control group, miR-449a group, miR-449b group,and anti-miR-449 group, the control group was cultured with osteogenic induction medium, and the remaining groups were cultured in osteogenic induction medium after transfection with miR-449a / b or inhibitors, respectively in culture 3, 6, 9, and12 d, the alkaline phosphatase activity was detected in each group, and the effect of miR-449a / b on the expression of osteogenic specific genes Runx2 and Osterix was detected by qRT-PCR. Results ①The fourth generation of mesenchymal stem cells was positive for CD44 and CD29, while the expression of CD34 and CD45 was negative, which is in line with the characteristics of bone marrow mesenchymal stem cells. ②After 3, 6, 9, 12 d of osteogenic induction culture, the alkaline phosphatase activity of MSCs cells increased compared with the previous time point, and it was time-dependent,the difference was statistically significant (P<0.05). After 3,6,9,12 d of osteogenic induction of MSCs, the expression levels of Runx2 and Osterix gradually increased. ③The AKP activity of miR-449a group and miR-449b group was lower than the control group, while the anti-miR-449 group increased AKP activity, the difference was statistically significant (P<0.05); miR-449a group and miR-449b group Runx2, Osterix expression was down-regulated compared with the control group,the difference was statistically significant (P<0.05). Conclusion This experiment successfully isolated and cultured bone marrow MSCs, which has the ability to differentiate into bone in vitro, and miR-449 can inhibit the differentiation of bone marrow MSCs into osteogenesis.

參考文獻/References:

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更新日期/Last Update: 2020-02-15
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