Li Xiang, Bai Yuanzhen, Tong Jia, Mao Guoqing
Nanjing University of Chinese Medicine, 210023, Nanjing, China.
Department of Orthopedics, Jiangsu Province Hospital of Chinese Medicine, Nanjing, China.
J Mol Histol. 2025 Apr 25;56(3):142. doi: 10.1007/s10735-025-10424-4.
Our paper aimed to disclose the effects of nobiletin (NOB) on hormone-induced osteoblast apoptosis and potential action mechanism. MC3T3-E1 cells were randomly separated into normal group, glucocorticoid (GC) group, L-NOB group, M-NOB group, H-NOB group, and Colivelin group (Colivelin: JAK2/STAT3 activator). CCK-8 was applied to ascertain the activity of MC3T3-E1 cells. FITC-Annexin V/PI method was applied to measure cell apoptosis. Alkaline phosphatase (ALP) assay kit was applied to measure ALP activity. Enzyme linked immunosorbent assay (ELISA) was implemented to ascertain the levels of IL-6, IL-1β, TNF-α, and ROS. Western blotting was implemented to distinguish the expressions of JAK2/STAT3 pathway proteins. The viability of MC3T3-E1 cells, ALP activities, and bcl-2 protein level were considerably decreased, while the apoptotic rate, the levels of TNF-α, IL-1β, IL-6, ROS, and the expressions of pJAK2/JAK2, pSTAT3/STAT3, Caspase-3, and bax proteins were greatly increased in each group after GC treatment. In comparison with GC group, MC3T3-E1 cell viability, ALP activity, and bcl-2 protein level in the L-NOB group, M-NOB group, and H-NOB group were greatly increased. Conversely, the apoptotic rate, the levels of TNF-α, IL-1β, IL-6, ROS, and the expressions of pJAK2/JAK2, pSTAT3/STAT3, Caspase-3, and bax proteins were markedly reduced. In contrast to H-NOB group, the apoptotic rate, the levels of TNF-α, IL-1β, IL-6, ROS, and the expressions of pJAK2/JAK2, pSTAT3/STAT3, Caspase-3, and bax proteins in Colivelin group were considerably enhanced, while MC3T3-E1 cell viability, ALP activity, and bcl-2 protein level were greatly declined. NOB ameliorates hormone-induced osteoblast apoptosis by reducing JAK2/STAT3 signaling activity.
我们的论文旨在揭示诺米林（NOB）对激素诱导的成骨细胞凋亡的影响及其潜在作用机制。将MC3T3-E1细胞随机分为正常组、糖皮质激素（GC）组、低剂量诺米林（L-NOB）组、中剂量诺米林（M-NOB）组、高剂量诺米林（H-NOB）组和可利韦林组（可利韦林：JAK2/STAT3激活剂）。采用CCK-8法检测MC3T3-E1细胞活性。采用FITC-Annexin V/PI法检测细胞凋亡。采用碱性磷酸酶（ALP）检测试剂盒检测ALP活性。采用酶联免疫吸附测定（ELISA）法检测IL-6、IL-1β、TNF-α和活性氧（ROS）水平。采用蛋白质免疫印迹法检测JAK2/STAT3信号通路蛋白的表达。GC处理后，各组MC3T3-E1细胞活力、ALP活性和bcl-2蛋白水平显著降低，而凋亡率、TNF-α、IL-1β、IL-6、ROS水平以及pJAK2/JAK2、pSTAT3/STAT3、Caspase-3和bax蛋白的表达显著增加。与GC组相比，L-NOB组、M-NOB组和H-NOB组的MC3T3-E1细胞活力、ALP活性和bcl-2蛋白水平显著升高。相反，凋亡率、TNF-α、IL-1β、IL-6、ROS水平以及pJAK2/JAK2、pSTAT3/STAT3、Caspase-3和bax蛋白的表达显著降低。与H-NOB组相比，可利韦林组的凋亡率、TNF-α、IL-1β、IL-6、ROS水平以及pJAK2/JAK2、pSTAT3/STAT3、Caspase-3和bax蛋白的表达显著增强，而MC3T3-E1细胞活力、ALP活性和bcl-2蛋白水平显著下降。诺米林通过降低JAK2/STAT3信号活性改善激素诱导的成骨细胞凋亡。
