摘要
目的探讨miR-129-5p靶向转录因子4(transcription factor 4,TCF4)对后纵韧带细胞(posterior longitudinal ligament cells,PLLCs)成骨分化的影响。方法选择2021年5月至2022年5月在中国人民解放军联勤保障部队第九六七医院接受手术治疗后纵韧带骨化(OPLL)患者和颈椎外伤患者各10例,分为OPLL组和PLL组,术中获取后纵韧带,应用贴壁培养的方法对两组的后纵韧带细胞进行体外培养,利用qRT-PCR和Western Blot方法检测miR-129-5p和TCF4对后纵韧带细胞成骨相关指标(RUNX2、ALP、OPN)表达情况的影响;碱性磷酸酶染色(ALP)和茜素红染色(ARS)检测miR-129-5p和TCF4对后纵韧带细胞碱性磷酸酶活性和钙沉积的影响;双荧光素酶实验验证miR-129-5p和TCF4之间的结合关系;miR-129-5p mimic和OE-TCF4共转染研究两者对后纵韧带细胞成骨分化的影响。结果miR-129-5p mimic和OE-TCF4能够促进后纵韧带细胞成骨相关基因的表达水平、碱性磷酸酶活性及钙结节沉积,miR-129-5p和TCF4两者之间存在靶向结合关系,且miR-129-5p能够靶向抑制TCF4表达,进而抑制后纵韧带细胞成骨相关基因的表达水平、碱性磷酸酶活性及钙结节沉积。结论miR-129-5p靶向抑制TCF4表达,进而抑制PLLCs的成骨分化。
Objective To investigate the effect of miR-129-5p targeting transcription factor 4(TCF4)on the osteogenic differentiation of posterior longitudinal ligament cells(PLLCs).Methods A total of 20 patients were selected,with 10 individuals diagnosed with ossification of the posterior longitudinal ligament(OPLL)and another 10 patients with cervical trauma to receive surgical treatment at No.967 Hospital of the Joint Logistic Support Force from May 2021 to May 2022,and were divided into OPLL group and PLL group.The posterior longitudinal ligament was obtained during the operation,and the PLLCs of the two groups were cultured in vitro by the method of adherent culture.To examine the effects of miR-129-5p and TCF4 on the expression of osteogenesis-related indicators(RUNX2,ALP,OPN)by qRT-PCR and Western Blot analysis.To evaluate the impact of miR-129-5p and TCF4 on calcium levels in PLLCs,Alkaline phosphatase staining(ALP)and Alizarin Red staining(ARS)techniques were employed.Furthermore,dual luciferase experiments were performed to verify the binding relationship between miR-129-5p and TCF4.In addition,co-transfection of miR-129-5p mimic and OE-TCF4 was performed to investigate their respective effects on the osteogenic differentiation of PLLCs.Results The findings demonstrated that miR-129-5p mimic and OE-TCF4 enhance the expression level of osteogenesis-related genes,calcium nodule deposition,and alkaline phosphatase activity in PLLCs.Additionally,a targeted binding relationship between miR-129-5p and TCF4 was observed,indicating that miR-129-5p can target and inhibit TCF4 expression,and then inhibit the expression level of osteogenesis-related genes,calcium nodule deposition,and alkaline phosphatase activity in PLLCs.Conclusion The study concludes that miR-129-5p targets and inhibits the expression of TCF4,which ultimately inhibits the osteogenic differentiation of PLLCs.
作者
张浩
唐晓丹
吕晶
孙浩
徐辰
Zhang Hao;Tang Xiaodan;Lyu Jing;Sun Hao;Xu Chen(Department of Orthopedics,No.967 Hospital of the Joint Logistic Support Force,Dalian Liaoning,116021;Department of Orthopedics,Changzheng Hospital Affiliated to Naval Medical University,Shanghai,200003,China)
出处
《生物骨科材料与临床研究》
2025年第1期1-6,共6页
Orthopaedic Biomechanics Materials and Clinical Study
基金
大连市医学科研计划项目(2212024)。
