Xie Tian, Yuan Jun, Mei Ling, Li Ping, Pan Ruijie
Department of Orthopedics, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, Hubei 430014, P.R. China.
College of Acupuncture and Bone Injury, Hubei University of Chinese Medicine, Wuhan, Hubei 430061, P.R. China.
Exp Ther Med. 2022 May 26;24(1):469. doi: 10.3892/etm.2022.11396. eCollection 2022 Jul.
Luteolin (3',4',5,7-tetrahydroxy flavone) is a flavonoid, which is widely distributed in various plants including flowers, vegetables, and medicinal herbs and spices. Luteolin can be applied in the treatment of various diseases due to its multiple biological activities, such as anti-inflammatory, anticancer, and antioxidative activity. However, its role in intervertebral disc degeneration has not been previously reported. Therefore, the purpose of the present study was to explore the effects of luteolin on Tumor necrosis factor (TNF)-α-induced inflammatory injury and senescence of human nucleus pulposus cells (HNPCs), as well as the underlying mechanisms of action of this compound. Cell viability and apoptosis were assessed by MTT assay and TUNEL staining, respectively. ELISA kits were applied to detect the levels of inflammatory cytokines and the activity of telomerase. Senescence β-galactosidase staining was used to detect the activity levels of β-galactosidase in the cells. Cell transfection was performed to achieve interference of sirtuin 6 (Sirt6). The protein expression levels were detected by western blot analysis. TUNEL staining and western blot analysis were performed to assess the expression levels of apoptosis-related proteins. The results indicated that TNF-α induced a significant decrease in HNPC viability and an increase in inflammatory factor levels, while the application of luteolin effectively increased cell viability and decreased intracellular interleukin (IL)-1β and IL-6 expression levels. Furthermore, luteolin decreased apoptosis compared with the TNF-α groups in a dose-dependent manner. In addition, the results of the detection kits suggested that luteolin reversed TNF-α-induced senescence. Notably, interference with Sirt6 partially reduced the protective effect of luteolin on TNF-α-induced HNPC senescence via the Sirt6/NF-κB pathway. In summary, the data indicated that luteolin suppresses TNF-α-induced inflammatory injury and senescence of HNPCs via the Sirt6/NF-κB pathway.
木犀草素(3',4',5,7 - 四羟基黄酮)是一种黄酮类化合物,广泛分布于各种植物中,包括花卉、蔬菜、药草和香料。由于木犀草素具有多种生物活性,如抗炎、抗癌和抗氧化活性,因此可用于治疗各种疾病。然而,其在椎间盘退变中的作用此前尚未见报道。因此,本研究的目的是探讨木犀草素对肿瘤坏死因子(TNF)-α诱导的人髓核细胞(HNPCs)炎症损伤和衰老的影响,以及该化合物的潜在作用机制。分别通过MTT法和TUNEL染色评估细胞活力和凋亡情况。应用ELISA试剂盒检测炎症细胞因子水平和端粒酶活性。采用衰老β - 半乳糖苷酶染色检测细胞中β - 半乳糖苷酶的活性水平。进行细胞转染以实现对沉默调节蛋白6(Sirt6)的干扰。通过蛋白质免疫印迹分析检测蛋白质表达水平。进行TUNEL染色和蛋白质免疫印迹分析以评估凋亡相关蛋白的表达水平。结果表明,TNF - α导致HNPC活力显著降低,炎症因子水平升高,而木犀草素的应用有效提高了细胞活力,并降低了细胞内白细胞介素(IL)-1β和IL - 6的表达水平。此外,与TNF - α组相比,木犀草素以剂量依赖性方式降低了细胞凋亡。另外,检测试剂盒的结果表明木犀草素可逆转TNF - α诱导的衰老。值得注意的是,干扰Sirt6通过Sirt6/NF - κB途径部分降低了木犀草素对TNF - α诱导的HNPC衰老的保护作用。综上所述,数据表明木犀草素通过Sirt6/NF - κB途径抑制TNF - α诱导的HNPC炎症损伤和衰老。