The antimicrobial peptide KR-12 promotes the osteogenic differentiation of human bone marrow stem cells by stimulating BMP/SMAD signaling.
The antimicrobial peptide KR-12 promotes the osteogenic differentiation of human bone marrow stem cells by stimulating BMP/SMAD signaling.
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抗菌肽KR-12通过刺激BMP/SMAD信号促进人骨髓干细胞成骨分化
DOI：
10.1039/c8ra00750k
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发表时间：
2018-04-23
期刊：
影响因子：
3.9
通讯作者：
Yue, Bing
中科院分区：
文献类型：
--
作者：
Li, Hui;Zhang, Shutao;Nie, Bin'en;Du, Zhe;Long, Teng;Yue, Bing
KR-12是人抗菌肽cathelicidin（LL-37）的最小片段，在治疗包括骨髓炎在内的多种感染中发挥重要作用。我们的前期工作发现，KR-12可促进人骨髓间充质干细胞（HBMSCs）的成骨分化。本研究探讨KR-12是否影响HBMSC成骨分化，以及所涉及的分子机制。细胞计数法检测KR-12对HBMSC增殖的影响，流式细胞仪检测KR-12对HBMSC细胞周期进程和凋亡的影响。采用碱性磷酸酶、天狼星红、茜素红染色及定量分析方法研究HBMSCs的成骨分化。实时荧光定量PCR检测成骨分化标志物的表达。通过荧光素酶报告基因测定和蛋白质印迹分析来检查潜在相关途径的激活。KR-12处理增加了HBMSC的成骨分化，无细胞毒性，并且不影响细胞周期或诱导细胞凋亡。荧光素酶报告基因分析表明，KR-12激活骨形态发生蛋白2（BMP 2）的转录，BMP/SMAD途径中的关键基因。Western blot分析表明，BMP/SMAD信号显着激活KR-12刺激成骨诱导条件。SMAD磷酸化被KR-12处理激活，并被转化生长因子-β/SMAD抑制剂（LDN-193189 HCL）和BMP 2小干扰RNA（si-BMP 2）抑制。LDN-193189 HCL和si-BMP 2处理也消除了KR-12诱导的HBMSC的成骨分化。总之，我们的研究结果表明，KR-12通过激活BMP/SMAD信号促进HBMSC成骨。
KR-12 is the smallest fragment of human antimicrobial peptide cathelicidin (LL-37), and could play key roles in the treatment of multiple infections, including osteomyelitis. Our preliminary work found that KR-12 enhances the osteogenic differentiation of human bone marrow mesenchymal stem cells (HBMSCs). The present study investigated whether KR-12 affects HBMSC osteogenic differentiation, as well as the molecular mechanisms involved. HBMSC proliferation in the presence of KR-12 was observed with a cell counting 8 assay, and its effects on HBMSC cell cycle progression and apoptosis were examined by flow cytometry. Alkaline phosphatase, Sirius Red, and Alizarin Red staining and quantitative assays were used to study the osteogenic differentiation of HBMSCs. The expression of osteogenic differentiation markers was detected by real-time quantitative PCR analysis. The activation of potentially related pathways was examined by luciferase reporter assay and western blot analysis. KR-12 treatment increased the osteogenic differentiation of HBMSCs without cytotoxicity and did not influence the cell cycle or induce apoptosis. Luciferase reporter assays showed that KR-12 activated the transcription of bone morphogenetic protein 2 (BMP2), a key gene in the BMP/SMAD pathway. Western blot analysis indicated that BMP/SMAD signaling was markedly activated by KR-12 stimulation in osteogenic induction conditions. SMAD phosphorylation was activated by KR-12 treatment, and was inhibited by both a transforming growth factor-β/SMAD inhibitor (LDN-193189 HCL) and BMP2 small interfering RNA (si-BMP2). LDN-193189 HCL and si-BMP2 treatment also abolished the KR-12-induced osteogenic differentiation of HBMSCs. In conclusion, our results suggest that KR-12 promotes HBMSC osteogenesis through the activation of BMP/SMAD signaling.
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影响因子：
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