Yang Yaming, Yan Zheng, Xie Qi, Wang Yong, Liu Zhiying, Lei Min
Department of Clinical Nutrition, The Third Hospital of Hebei Medical University, Shijiazhuang, Hebei 050051, China.
School of Public Health, Hebei Medical University, Shijiazhuang, Hebei 050017, China.
Aging (Albany NY). 2024 Apr 3;16(7):6334-6347. doi: 10.18632/aging.205708.
The purpose of this study is to observe LP45 ( 45) to investigate the mechanism by which LP45 attenuates oxidative stress-induced damage and regulates the osteoblast-osteoclast balance.
The oxidative stress level and osteoblast- and osteoclast-related proteins were detected by immunofluorescence staining, Western blotting, ROS fluorescent probe and ELISA. Osteoblast cell proliferation capacity was determined by the CCK-8 assay. X-ray observation and HE staining were used to detect the effect of LP45 on osteoporosis.
The expression level of SHP2 and Src was significantly increased, and the expression levels of NOX4, P22, P47, IL-1β, NLRP3, IRF3, RANK, β-catenin and INF-β were inhibited in LP45 group and LPS + LP45 group as compared to those in LPS group. Compared with that in LPS group, the concentration of SOD was increased and the concentration of MDA was decreased in LPS + LP45 group. The protein expressions of OPG, RANKL, RUNX3, RANK and β-catenin in LP45 group and LPS + LP45 group increased. The protein expressions of NF-κB, CREB and AP-1 in LP45 group and LPS + LP45 group decreased significantly. The results were also confirmed by immunofluorescence staining and ROS fluorescent probe. X-ray observation and HE staining showed that LP45 could inhibit the progression of osteoporosis.
LP45 can exert its antioxidant effect by inhibiting the production of oxidative stress to activate the SHP2 signaling pathway, thus promoting osteoblast differentiation and repressing osteoclast formation to maintain bone homeostasis and improve bone metabolism.
本研究旨在观察LP45，以探究LP45减轻氧化应激诱导的损伤并调节成骨细胞 - 破骨细胞平衡的机制。
通过免疫荧光染色、蛋白质印迹法、活性氧（ROS）荧光探针和酶联免疫吸附测定（ELISA）检测氧化应激水平以及与成骨细胞和破骨细胞相关的蛋白。采用细胞增殖-毒性检测试剂盒（CCK-8）法测定成骨细胞的增殖能力。通过X射线观察和苏木精-伊红（HE）染色检测LP45对骨质疏松症的影响。
与脂多糖（LPS）组相比，LP45组和LPS + LP45组中SHP2和Src的表达水平显著升高，而NOX4、P22、P47、白细胞介素-1β（IL-1β）、NLRP3、干扰素调节因子3（IRF3）、核因子κB受体活化因子（RANK）、β-连环蛋白和干扰素-β（INF-β）的表达水平受到抑制。与LPS组相比，LPS + LP45组中超氧化物歧化酶（SOD）浓度升高，丙二醛（MDA）浓度降低。LP45组和LPS + LP45组中骨保护素（OPG）、核因子κB受体活化因子配体（RANKL）、 runt相关转录因子3（RUNX3）、RANK和β-连环蛋白的蛋白表达增加。LP45组和LPS + LP45组中核因子κB（NF-κB）、环磷腺苷效应元件结合蛋白（CREB）和激活蛋白-1（AP-1）的蛋白表达显著降低。免疫荧光染色和ROS荧光探针也证实了上述结果。X射线观察和HE染色表明LP45可抑制骨质疏松症的进展。
LP45可通过抑制氧化应激的产生来发挥其抗氧化作用，从而激活SHP2信号通路，进而促进成骨细胞分化并抑制破骨细胞形成，以维持骨稳态并改善骨代谢。
