棕榈酸靶向调控趋化因子受体4促进骨髓间充质干细胞体外迁移的研究

Objective To study whether the effective component of traditional Chinese medicine safflower, palmitic acid, promotes the in vitro migration of rat bone marrow mesenchymal stem cells (rBMSCs) by targeting and regulating C - X - C chemokine receptor 4. Methods rBMSCs were cultured by the whole bone marrow adherent culture method, and the third generation (P3) rBMSCs were taken for experiments. Palmitic acid was used to intervene and treat rBMSCs according to the low, medium and high concentration gradients (5g/L, 10g/L, 20g/L), and Transwell experiments were carried out to determine that the optimal concentration for promoting migration was 10g/L. RT - qPCR was used to detect the effect of different concentrations of palmitic acid on the expression of CXCR4 mRNA. At the same time, CXCR4 siRNA plasmid was used to transfect rBMSCs, and they were divided into the palmitic acid group (10g/L), the CXCR4 siRNA group, the palmitic acid (10g/L) + CXCR4 siRNA group and the blank control group (10g/L). Further, through Transwell experiments, RT - qPCR experiments and Western Blot experiments, the effect of palmitic acid targeting and regulating C - X - C chemokine receptor 4 (CXCR4) on stem cell migration was verified. Results Different concentrations of palmitic acid could promote the migration of rBMSCs and up - regulate the expression of CXCR4 mRNA in rBMSCs, and the medium concentration of 10g/L was the best (P < 0.05). The Transwell experiment of rBMSCs after transfection with CXCR4 siRNA showed that the ability of palmitic acid to promote the migration of rBMSCs was related to the increase in the expression of C - X - C chemokine receptor 4. Conclusion Palmitic acid promotes the in vitro migration of rat bone marrow mesenchymal stem cells by up - regulating the expression of C - X - C chemokine receptor 4.

目的研究中药红花有效成分棕榈酸是否通过靶向调控C-X-C趋化因子受体4促进大鼠骨髓间充质干细胞(rBMSCs)体外迁移。方法通过全骨髓贴壁培养法培养rBMSCs,取第三代(P3)rBMSCs进行实验。棕榈酸按照低、中、高浓度梯度(5g/L、10g/L、20g/L)干预处理rBMSCs,并进行Transwell实验,确定最佳促迁移浓度为10g/L。应用RT-qPCR检测不同浓度的棕榈酸对CXCR4 mRNA表达的影响,同时运用CXCR4 siRNA质粒转染rBMSCs,并分为棕榈酸组(10g/ L)、CXCR4 siRNA组、棕榈酸(10g/L) +CXCR4 siRNA组及空白对照组(10g/L),进一步通过Transwell实验、RT-qPCR实验、Western Blot实验,验证棕榈酸靶向调控C-X-C趋化因子受体4(CXCR4)对干细胞迁移的影响。结果不同浓度的棕榈酸均可促进rBMSCs的迁移并上调rBMSCs的CXCR4 mRNA的表达,其中以中浓度10g/L最佳(P<0.05)。转染CXCR4 siRNA后rBMSCs的Transwell实验表明,棕榈酸促进rBMSCs迁移的能力与提高C-X-C趋化因子受体4表达相关。结论棕榈酸通过上调C-X-C趋化因子受体4的表达促进大鼠骨髓间充质干细胞体外迁移。