Lu Zhi-Jun, Gu Hou-Yun, Li Zhi-Qiang, Lin Fei-Xiang
Department of Spine Surgery, Ganzhou People's Hospital (The Affiliated Ganzhou Hospital of Jiangxi Medical College of Nanchang University, Ganzhou Hospital-Nanfang Hospital of Southern Medical University), Ganzhou, Jiangxi 341000, P.R. China.
Exp Ther Med. 2024 Oct 1;28(6):446. doi: 10.3892/etm.2024.12736. eCollection 2024 Dec.
The present study investigated the effect of connexin 43 (Cx43) on the regulation of osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (BMSCs) using low-frequency-pulsed electromagnetic fields (LPEMF). The BMSCs were isolated and cultured using adherent whole-bone marrow cultures. CCK-8 assay was used to detect the effects of LPEMF on the proliferation ability of BMSCs and alkaline phosphatase (ALP) activity and the levels of osteogenic marker genes were detected to evaluate the osteogenic ability change following LPEMF treatment. Lentiviral vector-mediated RNA interference was transfected into BMSCs to inhibit the expression of Cx43 and western blotting was used to detect Cx43 expression. The BMSCs showed the highest proliferation following LPEMF treatment at 80 Hz for 1 h. The results of ALP activity, osteogenic marker genes and Alizarin Red S staining showed that the osteogenic ability was notably increased following LPEMF treatment at 80 Hz for 1 h. Cx43 expression increased during the osteogenic differentiation of BMSCs following LPEMF treatment at 80 Hz. The enhanced osteogenic differentiation of the LPEMF-treated BMSCs were partially reversed when Cx43 expression was inhibited. LPEMF may promote the osteogenic differentiation of BMSCs by regulating Cx43 expression and enhancing osteogenic ability.
本研究利用低频脉冲电磁场（LPEMF），探讨了连接蛋白43（Cx43）对大鼠骨髓间充质干细胞（BMSCs）成骨分化调控的影响。采用贴壁全骨髓培养法分离培养BMSCs。采用CCK-8法检测LPEMF对BMSCs增殖能力的影响，检测碱性磷酸酶（ALP）活性及成骨标志基因水平，以评估LPEMF处理后成骨能力的变化。将慢病毒载体介导的RNA干扰转染至BMSCs中以抑制Cx43的表达，并采用蛋白质印迹法检测Cx43表达。BMSCs在80 Hz的LPEMF处理1 h后增殖最为显著。ALP活性、成骨标志基因及茜素红S染色结果显示，80 Hz的LPEMF处理1 h后成骨能力显著增强。在80 Hz的LPEMF处理后，BMSCs成骨分化过程中Cx43表达增加。当Cx43表达受到抑制时，LPEMF处理的BMSCs增强的成骨分化被部分逆转。LPEMF可能通过调节Cx43表达和增强成骨能力来促进BMSCs的成骨分化。
