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首页> 外文期刊>Drug Design, Development and Therapy >N -Acetyl-l-Leucine-Polyethyleneimine-Mediated Delivery of CpG Oligodeoxynucleotides 2006 Inhibits RAW264.7 Cell Osteoclastogenesis
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N -Acetyl-l-Leucine-Polyethyleneimine-Mediated Delivery of CpG Oligodeoxynucleotides 2006 Inhibits RAW264.7 Cell Osteoclastogenesis

机译：N-乙酰-1-亮氨酸 - 聚乙烯亚胺介导的CPG寡脱氧核苷酸2006的递送抑制RAW264.7细胞骨质细胞发生

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Introduction: CpG oligodeoxynucleotides (CpG ODN) play important roles in resisting inflammation and bone resorption. However, the inherent instability and rapid degradation hinder their wider application. This study aimed to evaluate whether N -acetyl-L-leucine-modified polyethyleneimine ( N -Ac-L-Leu-PEI) could effectively deliver CpG ODN 2006 to RAW264.7 cells and and if it can regulate osteoclastogenesis in vitro. Materials and Methods: Gel retardation assay was conducted to evaluate whether N - Ac-L-Leu-PEI and CpG ODN could form a stable complex. RAW264.7 cells were divided into four groups of control group, ODN group, phosphorothioate ODN group and N -Ac-L-Leu-PEI/ODN group. Fluorescence assay was conducted to evaluate the transfection rate of ODNs in different groups. Cell viability was determined by MTT assay. Cell apoptosis was determined by live-dead cell staining and flow cytometry assay. Relative expression levels of osteoclastic differentiation factors, including Nfatc, c-fos, receptor activator of nuclear factor κB (RANK), and matrix metalloproteinase 9 (MMP9), were determined by real-time PCR and Western blot. Results: N -Ac-L-Leu-PEI and CpG ODN could form a stable complex at a mass ratio of 1:1 (w:w). MTT assay showed that the cell viability of N -Ac-L-Leu-PEI was relatively high even at a mass ratio of 8 μg/mL. The transfection rate of N -Ac-L-Leu-PEI-ODN complex was higher than 90%. The cell proliferation and apoptosis was significantly enhanced in N -Ac-L-Leu-PEI- CpG ODN group when compared to those in phosphorothioate CpG ODN. The expression levels of Nfatc, c-fos, RANK, and MMP9 were significantly decreased in N -Ac-L-Leu-PEI/ODN complex group. Discussion: N -Ac-L-Leu-PEI could be a potential gene vehicle for the prevention of periodontitis-mediated bone resorption.
机译：简介：CPG寡脱氧核苷酸（CPG ODN）在抵抗炎症和骨吸收中起重要作用。然而，固有的不稳定性和快速降解阻碍了他们更广泛的应用。该研究旨在评估N-乙酰基-L-亮氨酸改性的聚乙烯（N-CAC-L-Leu-PEI）是否可以有效地将CPG ODN 2006递送至RAW264.7细胞，并且如果它可以在体外调节骨核细胞发生。材料和方法：进行凝胶延迟测定以评估N - AC-L-Leu-PEI和CpG ODN是否可以形成稳定的复合物。将Raw264.7细胞分为四组对照组，ODN组，硫代磷酸ODN基团和N-Ac-L-Leu-PeI / ODN组。进行荧光测定以评估不同组中ODN的转染率。通过MTT测定法测定细胞活力。细胞凋亡由活体 - 死细胞染色和流式细胞术测定。通过实时PCR和Western印迹测定骨细胞分化因子的相对表达水平，包括NFATC，C-FOS，核因子κB（秩序）和基质金属蛋白酶9（MMP9）的蛋白酶9（MMP9）。结果：N -Ac-L-Leu-PEI和CPG ODN可以以质量比为1：1（W：W）形成稳定的复合物。 MTT测定表明，即使以8μg/ ml的质量比，N -Ac-L-Leu-Pei的细胞活力也相对较高。 N -Ac-L-Leu-PEI-ODN复合物的转染率高于90％。与硫代磷酸盐磷酸盐胺odN的那些相比，在N -Ac-L-Leu-PEI-CPG ODN组中显着增强了细胞增殖和细胞凋亡。 NFATC，C-FOS，等级和MMP9的表达水平在N-AC-L-Leu-PEI / ODN复合物中显着降低。讨论：N -Ac-L-Leu-PEI可以是预防牙周炎介导的骨吸收的潜在基因载体。

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