Qiu Chen, Li Zhaowen, Peng Puji
Department of Sports Medicine, The Affiliated Hospital of Wuhan Sports University, Wuhan, 430000, China.
Department of Orthopedics, Henan Provincial People's Hospital, Zhengzhou, 450003, China.
Regen Ther. 2024 Mar 5;27:1-11. doi: 10.1016/j.reth.2024.02.009. eCollection 2024 Dec.
To investigate the protective effect human umbilical cord mesenchymal stem cells (hUC-MSCs) have on Dexamethasone (Dex)-induced apoptosis in osteogenesis via the Nrf2-ARE signaling pathway.
Glucocorticoid-induced osteonecrosis of the femoral head (GC-ONFH) was developed in rats through the administration of lipopolysaccharide and methylprednisolone. The incidence of femoral head necrosis, cavity notch, apoptosis of osteoblasts, and bone density were observed by HE staining, TUNEL staining, and Micro-CT. HUC-MSCs were co-cultured with mouse pre-osteoblast MC3T3-E1. The survival rate of osteoblasts was determined by CCK8, and apoptosis and ROS levels of osteoblasts were determined by flow cytometer. The viability of antioxidant enzymes SOD, GSH-Px, and CAT was analyzed by biochemistry. Nrf2 expression levels and those of its downstream proteins and apoptosis-related proteins were analyzed by Western blotting.
In rats, hUC-MSCs can reduce the rates of empty bone lacuna and osteoblast apoptosis that are induced by glucocorticoids (GCs), while reducing the incidence of GC-ONFH. hUC-MSCs can significantly improve the survival rate and antioxidant SOD, GSH-Px, and CAT activity of MC3T3-E1 cells caused by Dex, and inhibit apoptosis and oxidative stress levels. In addition, hUC-MSCs can up-regulate the expression of osteoblast antioxidant protein Nrf2 and its downstream protein HO-1, NQO-1, GCLC, GCLM, and apoptosis-related protein bcl-2, while also down-regulating the expression of apoptosis-related protein bax, cleaved caspase-3, cleaved caspase-9, and cytochrome C in MC3T3-E1 cells. hUC-MSCs improve the ability of MC3T3-E1 cells to mineralize to osteogenesis. However, the promoting effects of hUC-MSCs were abolished following the blocking of the Nrf2-ARE signaling pathway for osteoblasts.
The results reveal that hUC-MSCs can reduce Dex-induced apoptosis in osteoblasts via the Nrf2-ARE signaling pathway.
探讨人脐带间充质干细胞（hUC-MSCs）通过Nrf2-ARE信号通路对成骨过程中地塞米松（Dex）诱导的细胞凋亡的保护作用。
通过给予大鼠脂多糖和甲基强的松龙建立糖皮质激素诱导的股骨头坏死（GC-ONFH）模型。通过苏木精-伊红（HE）染色、TUNEL染色和显微CT观察股骨头坏死的发生率、空洞缺口、成骨细胞凋亡和骨密度。将hUC-MSCs与小鼠前成骨细胞MC3T3-E1共培养。采用CCK8法检测成骨细胞的存活率，采用流式细胞仪检测成骨细胞的凋亡率和活性氧（ROS）水平。通过生化分析抗氧化酶超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）和过氧化氢酶（CAT）的活性。采用蛋白质免疫印迹法分析Nrf2及其下游蛋白和凋亡相关蛋白的表达水平。
在大鼠中，hUC-MSCs可降低糖皮质激素（GCs）诱导的空骨陷窝率和成骨细胞凋亡率，同时降低GC-ONFH的发生率。hUC-MSCs可显著提高Dex诱导的MC3T3-E1细胞的存活率以及抗氧化酶SOD、GSH-Px和CAT的活性，并抑制细胞凋亡和氧化应激水平。此外，hUC-MSCs可上调成骨细胞抗氧化蛋白Nrf2及其下游蛋白血红素加氧酶-1（HO-1）、醌氧化还原酶1（NQO-1）、谷氨酸半胱氨酸连接酶催化亚基（GCLC）、谷氨酸半胱氨酸连接酶调节亚基（GCLM）和凋亡相关蛋白bcl-2的表达，同时下调MC3T3-E1细胞中凋亡相关蛋白bax、裂解的半胱天冬酶-3（cleaved caspase-3）、裂解的半胱天冬酶-9（cleaved caspase-9）和细胞色素C的表达。hUC-MSCs提高了MC3T3-E1细胞向成骨细胞矿化的能力。然而，在阻断成骨细胞的Nrf2-ARE信号通路后，hUC-MSCs的促进作用被消除。
结果表明，hUC-MSCs可通过Nrf2-ARE信号通路减少Dex诱导的成骨细胞凋亡。
