Wang Lili, Ma Rufeng, Guo Yubo, Sun Jing, Liu Haixia, Zhu Ruyuan, Liu Chenyue, Li Jun, Li Lin, Chen Beibei, Sun Liping, Tang Jinfa, Zhao Dandan, Mo Fangfang, Niu Jianzhao, Jiang Guangjian, Fu Min, Brömme Dieter, Zhang Dongwei, Gao Sihua
Cell and Biochemistry Lab, Preclinical Medicine School, Beijing University of Chinese MedicineBeijing, China.
Chinese Material Medica School, Beijing University of Chinese MedicineBeijing, China.
Front Pharmacol. 2017 May 22;8:266. doi: 10.3389/fphar.2017.00266. eCollection 2017.
This study is designed to explore whether (FLL) exhibits antioxidant effect in ovariectomized (OVX) rats, and to identify the signaling pathway involved in this process. OVX rats were treated with FLL aqueous extract (3.5 g/kg) for 12 weeks. Serum, uteri, and tibias were harvested from the rats and the levels of total antioxidant capacity (TAC), nitric oxide (NO), malondialdehyde (MDA), 8-hydroxy-desoxyguanosine (8-OHdG), and superoxide dismutase (SOD) were determined. Changes in the levels of NF-κB-p65, phosphorylation of NF-κB-p65 (NF-κB-pp65), NF-κB inhibitor alpha (IκBα), phosphorylation of IκBα (p-IκBα), and NADPH oxidase 4 (Nox4) in uteri and tibias were determined by western blot, immunofluorescent and immunohistochemical analysis, respectively. In addition, the expression of cytochrome C (Cyto-C) and B-cell lymphoma-2 (Bcl-2) were determined in the tibias of rats. Histopathological changes in the bones were evaluated by hematoxylin-eosin staining. Bone mineral density (BMD) was determined in rat femurs by dual X-ray absorptiometry. Treatment of OVX rats with FLL aqueous extract improved redox homeostasis by increasing the levels of TAC and NO as well as decreasing the levels of MDA and 8-OHdG in serum, tibias, and uteri. Further, FLL extract also downregulated the expression of Nox4, NF-κB-p65, NF-κB-pp65, and p-IκBα in the uteri and tibias. Furthermore, administration of FLL-OVX rats increased Bcl-2 expression and prevented cytoplasmic release of mitochondrial Cyto-C in the tibias. In addition, FLL treatment also improved bone microstructure and increased cortical bone thickness as well as increased BMD values in the femurs of OVX rats. FLL treatment may suppress oxidative stress response in OVX rats via regulating the Nox4/ROS/NF-κB signaling pathway. These results suggest the potential of using FLL as a natural antioxidant agent in preventing the development of osteoporosis.
本研究旨在探讨（FLL）在去卵巢（OVX）大鼠中是否具有抗氧化作用，并确定参与该过程的信号通路。将OVX大鼠用FLL水提取物（3.5 g/kg）处理12周。从大鼠身上采集血清、子宫和胫骨，测定总抗氧化能力（TAC）、一氧化氮（NO）、丙二醛（MDA）、8-羟基脱氧鸟苷（8-OHdG）和超氧化物歧化酶（SOD）的水平。分别通过蛋白质免疫印迹、免疫荧光和免疫组织化学分析测定子宫和胫骨中NF-κB-p65、NF-κB-p65磷酸化（NF-κB-pp65）、NF-κB抑制剂α（IκBα）、IκBα磷酸化（p-IκBα）和NADPH氧化酶4（Nox4）水平的变化。此外，还测定了大鼠胫骨中细胞色素C（Cyto-C）和B细胞淋巴瘤-2（Bcl-2）的表达。通过苏木精-伊红染色评估骨骼的组织病理学变化。用双能X线吸收法测定大鼠股骨的骨密度（BMD）。用FLL水提取物处理OVX大鼠可通过提高血清、胫骨和子宫中TAC和NO的水平以及降低MDA和8-OHdG的水平来改善氧化还原稳态。此外，FLL提取物还下调了子宫和胫骨中Nox4、NF-κB-p65、NF-κB-pp65和p-IκBα的表达。此外，给FLL-OVX大鼠给药可增加Bcl-2表达并防止胫骨中线粒体Cyto-C的细胞质释放。此外，FLL处理还改善了骨微结构，增加了皮质骨厚度，并提高了OVX大鼠股骨的BMD值。FLL处理可能通过调节Nox4/ROS/NF-κB信号通路抑制OVX大鼠的氧化应激反应。这些结果表明FLL作为一种天然抗氧化剂在预防骨质疏松症发展方面的潜力。
