| 摘要: |
| ［摘要］ 目的 探讨力生长因子（MGF）对软骨终板干细胞（CESCs）增殖、迁移和分化的影响及其作用机制。方法 招募因退行性椎间盘疾病于广西壮族自治区人民医院进行椎间盘融合手术患者，术中采集其椎间盘软骨终板组织约2 cm3，取CESCs并进行培养。通过CCK-8实验评估MGF对CESCs增殖的影响；通过Transwell实验评估MGF对CESCs迁移和侵袭能力的影响；通过逆转录聚合酶链式反应（RT-PCR）和Western blot实验检测成骨、成软骨及成脂分化相关因子的表达水平，并分析细胞外信号调节激酶（ERK）磷酸化对MGF作用效果的影响。结果 CCK-8实验结果显示，MGF可促进CESCs增殖，且呈剂量依赖性。Transwell实验结果显示，MGF可促进CESCs迁移。经ERK抑制剂PD98059干预后，MGF的促增殖作用显著降低（P<0.05）。经胰岛素样生长因子-1受体（IGF-1R）抑制剂PQ401干预后，MGF的促迁移作用也显著降低（P<0.05）。RT-PCR检测结果显示，MGF组碱性磷酸酶（ALP）、Runt相关转录因子2（Runx2）和骨钙素（OC）的mRNA表达水平显著低于对照组（P<0.05），聚集蛋白聚糖（AGG）、性别决定区相关转录因子-9（SOX-9）和Ⅱ型胶原（CⅡ）的mRNA表达水平显著高于对照组（P<0.05）。Western blot实验结果也显示，MGF组ALP、Runx2和OC的蛋白表达水平显著低于对照组（P<0.05），AGG、SOX-9和CⅡ的蛋白表达水平显著高于对照组（P<0.05）。MGF组磷酸化细胞外信号调节激酶（p-ERK）/总细胞外信号调节激酶（t-ERK）比值显著高于对照组（P<0.05），MGF+PQ401组p-ERK/t-ERK比值低于MGF组，但仍高于对照组，差异有统计学意义（P<0.05）。结论 MGF通过诱导ERK磷酸化促进CESCs的增殖和迁移。 |
| 关键词: 软骨终板干细胞 力生长因子 增殖 迁移 细胞外信号调节激酶 胰岛素样生长因子-1受体 |
| DOI：10.3969/j.issn.1674-3806.2025.01.07 |
| 分类号:R 681.5 |
| 基金项目:广西科技计划项目（编号：桂科AD21220134）；广西自然科学基金项目（编号：2023GXNSFAA026285） |
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| Effects of mechano-growth factor on proliferation, migration and differentiation of cartilage endplate-derived stem cells and their acting mechanisms |
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LI Haoxi, WU Chunlan, PENG Guanlu, MO Kaiqi, YAO Shuyu, YU Chengqiang, LI Zhuhai, WEI Jianxun
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Department of Spine Surgery, the People′s Hospital of Guangxi Zhuang Autonomous Region(Guangxi Academy of Medical Sciences), Nanning 530021, China
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| Abstract: |
| ［Abstract］ Objective To explore the effects of mechano-growth factor(MGF) on proliferation, migration and differentiation of cartilage endplate-derived stem cells(CESCs) and their acting mechanisms. Methods The patients with degenerative disc disease who underwent disc fusion surgery in the People′s Hospital of Guangxi Zhuang Autonomous Region were recruited. During the surgery, about 2 cm3 of the patients′ cartilage endplate tissues of intervertebral disc were collected, and CESCs were collected and cultured. The effects of MGF on the proliferation of CESCs were evaluated by Cell Counting Kit-8(CCK-8) assay. The effects of MGF on the migration and invasion abilities of CESCs were evaluated by Transwell experiment. The expression levels of osteogenic, chondrogenic and lipogenic differentiation-related factors were detected by using reverse transcription-polymerase chain reaction(RT-PCR) and Western blot assay, and the influence of phosphorylated extracellular signal-regulated kinase(ERK) on the acting effect of MGF was analyzed. Results The results of CCK-8 assay showed that MGF could promote the proliferation of CESCs in a dose-dependent manner. The results of Transwell experiment showed that MGF could promote the migration of CESCs. After the intervention of ERK inhibitor PD98059, the proliferative effect of MGF was significantly weakened(P<0.05). After the intervention of insulin-like growth factor-1 receptor(IGF-1R) inhibitor PQ401, the promoting migration effect of MGF was also significantly weakened(P<0.05). The results of RT-PCR detection showed that the mRNA expression levels of alkaline phosphatase(ALP), Runt-related transcription factor 2(Runx2) and osteocalcin(OC) in the MGF group were significantly lower than those in the control group(P<0.05), while the mRNA expression levels of aggrecan(AGG), SRY-related high-mobility group box gene-9(SOX-9) and type Ⅱ collagen(CⅡ) in the MGF group were significantly higher than those in the control group(P<0.05). The results of Western blot assay also showed that the protein expression levels of ALP, Runx2 and OC in the MGF group were significantly lower than those in the control group(P<0.05), while the protein expression levels of AGG, SOX-9 and CⅡ in the MGF group were significantly higher than those in the control group(P<0.05). The ratio of phosphorylated ERK(p-ERK) to total extracellular signal-regulated kinase(t-ERK) in the MGF group was significantly higher than that in the control group(P<0.05), and the ratio of p-ERK to t-ERK in the MGF+PQ401 group was lower than that in the MGF group, but the ratio of p-ERK to t-ERK in the MGF+PQ401 group was still higher than that in the control group, and the differences were statistically significant(P<0.05). Conclusion MGF promotes the proliferation and migration of CESCs by inducing ERK phosphorylation. |
| Key words: Cartilage endplate-derived stem cells(CESCs) Mechano-growth factor(MGF) Proliferation Migration Extracellular signal-regulated kinase(ERK) Insulin-like growth factor-1 receptor( IGF-1R) |
