Tong Xishuai, Zhang Ying, Zhao Yutian, Li Yawen, Li Tan, Zou Hui, Yuan Yan, Bian Jianchun, Liu Zongping, Gu Jianhong
Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Institutes of Agricultural Science and Technology Development, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China.
Animals (Basel). 2023 Aug 7;13(15):2544. doi: 10.3390/ani13152544.
Vitamin D is a lipid soluble vitamin that is mostly used to treat bone metabolism-related diseases. In this study, the effect of Cd toxicity in vitro on osteogenic differentiation derived from BMSCs and the alleviating effect of lα, 25-(OH)D were investigated. Cell index in real time was monitored using a Real-time cell analyzer (RTCA) system. The activity of alkaline phosphatase (ALP), and the calcified nodules and the distribution of Runx2 protein were detected using ALP staining, alizarin red staining, and immunofluorescence, respectively. Furthermore, the mitochondrial membrane potential and the apoptotic rate of BMSCs, the mRNA levels of and type Ⅰ collagen alpha2 () genes, and the protein expression of Col1 and Runx2 were detected using flow cytometry, qRT-PCR and western blot, respectively. The proliferation of BMSCs and osteogenic differentiation were enhanced after treatment with different concentrations of lα, 25-(OH)D compared with the control group. However, 5 μmol/L Cd inhibited the proliferation of BMSCs. In addition, 10 nmol/L lα,25-(OH)D attenuated the toxicity and the apoptosis of BMSCs treated by Cd, and also promoted the osteogenic differentiation including the activity of ALP, and the protein expression of Col1 and Runx2. lα, 25-(OH)D can alleviate cadmium-induced osteogenic toxicity in White Leghorn chickens in vitro.
维生素D是一种脂溶性维生素,主要用于治疗与骨代谢相关的疾病。在本研究中,研究了体外镉毒性对骨髓间充质干细胞(BMSCs)成骨分化的影响以及1α,25-(OH)D的缓解作用。使用实时细胞分析仪(RTCA)系统监测实时细胞指数。分别采用碱性磷酸酶(ALP)染色、茜素红染色和免疫荧光检测ALP活性、钙化结节以及Runx2蛋白的分布。此外,分别采用流式细胞术、qRT-PCR和蛋白质免疫印迹法检测BMSCs的线粒体膜电位和凋亡率以及Ⅰ型胶原蛋白α2(Col1α2)基因的mRNA水平,以及Col1和Runx2的蛋白表达。与对照组相比,不同浓度的1α,25-(OH)D处理后BMSCs的增殖和成骨分化增强。然而,5 μmol/L镉抑制了BMSCs的增殖。此外,10 nmol/L 1α,25-(OH)D减轻了镉处理的BMSCs的毒性和凋亡,还促进了成骨分化,包括ALP活性以及Col1和Runx2的蛋白表达。1α,25-(OH)D可在体外减轻镉诱导的白来航鸡成骨毒性。