Department of Orthopedics, The First Affiliated Hospital of Bengbu Medical College; Anhui Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, China.
Drug Dev Res. 2021 Nov;82(7):1044-1054. doi: 10.1002/ddr.21811. Epub 2021 Mar 13.
Neobavaisoflavone (NBIF) is a flavonoid, which has a variety of pharmacological activities. However, the mechanism of NBIF in the treatment of osteoporosis still needs further exploration. The differentiation of osteoblast MC-3T3-E1 cells after treatment was observed by Alizarin red staining. Cell counting kit-8 and flow cytometry were used to detect viability, apoptosis, and reactive oxygen species (ROS) levels of treated MC-3T3-E1 cells, respectively. Malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were tested by ELISA kits. The expressions of lncRNA MALAT1, MEG3, CRNDE, Runx2, osteocalcin (OCN), osteopontin (OPN), collagen I (col-I), nuclear Nrf2, cytoplasm Nrf2, heme oxygenase-1 (HO-1) and quinone oxidoreductase 1 (NQO1) in treated MC-3T3-E1 cells were examined by Quantitative real-time PCR or Western blot. Dexamethasone (Dex) inhibited the viability of MC-3T3-E1 cells, while the appropriate amount of NBIF had no significantly effect on cell viability. Dex downregulated CRNDE expression, whereas NBIF upregulated CRNDE. Overexpressed CRNDE and NBIF reversed the inhibitory effects of Dex on cell viability, differentiation and levels of SOD, GSH-Px, Runx2, OCN, OPN, col-I, nuclear Nrf2, HO-1 and NQO1 while reversing the promoting effect of Dex on apoptosis and the levels of ROS, MDA, LDH and cytoplasm Nrf2 in MC-3T3-E1 cells, respectively, but shCRNDE further reversed the effects of NBIF in MC-3T3-E1 cells. NBIF protected osteoblasts from Dex-induced oxidative stress by upregulating the CRNDE-mediated Nrf2/HO-1 signaling pathway.
新巴西紫檀黄酮（NBIF）是一种黄酮类化合物，具有多种药理活性。然而，NBIF 治疗骨质疏松症的机制仍需要进一步探索。通过茜素红染色观察处理后成骨细胞 MC-3T3-E1 细胞的分化。分别用细胞计数试剂盒-8 和流式细胞术检测处理后的 MC-3T3-E1 细胞的活力、凋亡和活性氧（ROS）水平。通过 ELISA 试剂盒检测丙二醛（MDA）、乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）。用定量实时 PCR 或 Western blot 检测处理后的 MC-3T3-E1 细胞中长链非编码 RNA MALAT1、MEG3、CRNDE、Runx2、骨钙素（OCN）、骨桥蛋白（OPN）、I 型胶原（col-I）、核 Nrf2、细胞质 Nrf2、血红素加氧酶-1（HO-1）和醌氧化还原酶 1（NQO1）的表达。地塞米松（Dex）抑制 MC-3T3-E1 细胞活力，而适量 NBIF 对细胞活力无明显影响。Dex 下调 CRNDE 表达，而 NBIF 上调 CRNDE。过表达 CRNDE 和 NBIF 逆转了 Dex 对细胞活力、分化和 SOD、GSH-Px、Runx2、OCN、OPN、col-I、核 Nrf2、HO-1 和 NQO1 水平的抑制作用，同时逆转了 Dex 对 MC-3T3-E1 细胞中 ROS、MDA、LDH 和细胞质 Nrf2 水平的促进作用，但 shCRNDE 进一步逆转了 NBIF 在 MC-3T3-E1 细胞中的作用。NBIF 通过上调 CRNDE 介导的 Nrf2/HO-1 信号通路，保护成骨细胞免受 Dex 诱导的氧化应激。
