Department of Spine Surgery, Anhui Wanbei Coal-Electricity Group General Hospital, Suzhou City, Anhui Province, China.
Fourth Central Clinical School, Tianjin Medical University, Tianjin City, China.
J Physiol Pharmacol. 2024 Apr;75(2):173-183. doi: 10.26402/jpp.2024.2.06. Epub 2024 May 6.
Quercetin is widely distributed in plants as a flavonol compound with multiple biological activities. It has been found that quercetin can regulate bone homeostasis through multiple pathways and targets. This study investigated the role and specific molecular mechanisms of quercetin in regulating osteoblast viability, proliferation, migration and osteogenic differentiation. A mouse model of traumatic fracture was established and then 100 mg/kg quercetin corn oil suspension was gavaged at the same time every day for 28 days. miR-6089 and E2F transcription factor 2 (E2F2) expression levels in mice were measured. Fracture healing in mice was observed. MC3T3-E1 cells were transfected with plasmids targeting miR-6089 and E2F2, and cell viability, proliferation, migration, apoptosis, and osteogenic differentiation were determined. The targeting relationship between miR-6089 and E2F2 was verified. In vivo experiments showed that quercetin significantly increased osteocalcin (OCN) expression (P<0.05) and promoted fracture healing in traumatic fracture (TF) mice. miR-6089 expression was down-regulated (P<0.05) and E2F2 expression was up-regulated (P<0.05) in TF mice. Quercetin promoted miR-6089 expression and inhibited E2F2 expression (both P<0.05). In vitro results showed that quercetin promoted miR-6089 expression and inhibited E2F2 expression in a dose-dependent manner (both P<0.05). Quercetin dose-dependently promoted MC3T3-E1 cell viability, proliferation, migration, and osteogenic differentiation, and inhibited MC3T3-E1 cell apoptosis (all P<0.05). Up-regulating miR-6089 further promoted MC3T3-E1 cell viability, proliferation, migration and osteogenic differentiation, and inhibited MC3T3-E1 cell apoptosis (all P<0.05). miR-6089 targeted and regulated E2F2 expression. Up-regulating E2F2 attenuated the promoting effect of up-regulated miR-6089 on MC3T3-E1 cell viability, proliferation, migration, osteogenic differentiation, and inhibition of apoptosis (all P<0.05). We conclude that quercetin enhances osteoblast viability, proliferation, migration, and osteogenic differentiation by modulating the miR-6089/E2F2 axis, thereby promoting fracture healing.
槲皮素广泛存在于植物中，是一种具有多种生物学活性的类黄酮化合物。研究发现，槲皮素可以通过多种途径和靶点调节骨稳态。本研究探讨了槲皮素在调节成骨细胞活力、增殖、迁移和成骨分化中的作用及其特定分子机制。建立了创伤性骨折小鼠模型，然后每天同时给予 100mg/kg 槲皮素玉米油混悬液灌胃 28 天。测量小鼠 miR-6089 和 E2F 转录因子 2(E2F2)的表达水平。观察小鼠骨折愈合情况。用针对 miR-6089 和 E2F2 的质粒转染 MC3T3-E1 细胞，测定细胞活力、增殖、迁移、凋亡和成骨分化。验证 miR-6089 与 E2F2 的靶向关系。体内实验表明，槲皮素显著增加骨钙素(OCN)的表达(P<0.05)，促进创伤性骨折(TF)小鼠骨折愈合。TF 小鼠 miR-6089 表达下调(P<0.05)，E2F2 表达上调(P<0.05)。槲皮素促进 miR-6089 表达，抑制 E2F2 表达(均 P<0.05)。体外结果表明，槲皮素呈剂量依赖性地促进 miR-6089 表达，抑制 E2F2 表达(均 P<0.05)。槲皮素呈剂量依赖性地促进 MC3T3-E1 细胞活力、增殖、迁移和成骨分化，抑制 MC3T3-E1 细胞凋亡(均 P<0.05)。上调 miR-6089 进一步促进 MC3T3-E1 细胞活力、增殖、迁移和成骨分化，抑制 MC3T3-E1 细胞凋亡(均 P<0.05)。miR-6089 靶向调控 E2F2 表达。上调 E2F2 减弱上调 miR-6089 对 MC3T3-E1 细胞活力、增殖、迁移、成骨分化和抑制凋亡的促进作用(均 P<0.05)。综上所述，槲皮素通过调节 miR-6089/E2F2 轴增强成骨细胞活力、增殖、迁移和成骨分化，从而促进骨折愈合。
