Hui Wang1, Linhong Yuan1, Weiwei Ma1, Jing Han1, Yanhui Lu1, Lingli Feng1, Rong Xiao2. 1. School of Public Health, Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing 100069, China. 2. School of Public Health, Beijing Key Laboratory of Environmental Toxicology, Capital Medical University, Beijing 100069, China. Electronic address: xiaor22@ccmu.edu.cn.
Abstract
BACKGROUND: Diverse physiological and pathological functions of 27-hydroxycholesterol (27-OHC) were proved. However, cytotoxicity and potential influence of 27-OHC on cholesterol metabolism in neurons are unclear. DESIGN AND METHODS: In the vitro co-culture system, SH-SY5Y cells and C6 cells were applied to explore the potential cytotoxicity of 27-OHC. MTT assay was used to detect the cell proliferation. Cell vitality was measured by using the Annexin-FITC/PI test. Immunofluorescence technique was applied to observe the changes of mitochondria membrane potential. The expression of mRNA and protein (including SREBP-1, HMGCR, LXR-α, ABCA1) were measured by real-time PCR and western blot method respectively. RESULTS: 27-OHC induced apoptosis in co-cultured SH-SY5Y cells and C6 cells. 27-OHC treatment significantly inhibited cell viability and proliferation (p<0.05). Compared with control group, 27-OHC caused the reduction of mitochondrial membrane potential (p<0.05). Additionally, the mRNA and protein expression of cholesterol synthesis-related factors, such as SREBP-1, HMGCR, were down-regulated (p<0.05), while the mRNA and protein expression of cholesterol transport-related factors (LXR-α, ABCA1) were up-regulated (p<0.05). CONCLUSIONS: Cytotoxicity and cholesterol metabolism disorder induced by 27-OHC may contribute to the pathogenesis of neurodegenerative disease.
BACKGROUND: Diverse physiological and pathological functions of 27-hydroxycholesterol (27-OHC) were proved. However, cytotoxicity and potential influence of 27-OHC on cholesterol metabolism in neurons are unclear. DESIGN AND METHODS: In the vitro co-culture system, SH-SY5Y cells and C6 cells were applied to explore the potential cytotoxicity of 27-OHC. MTT assay was used to detect the cell proliferation. Cell vitality was measured by using the Annexin-FITC/PI test. Immunofluorescence technique was applied to observe the changes of mitochondria membrane potential. The expression of mRNA and protein (including SREBP-1, HMGCR, LXR-α, ABCA1) were measured by real-time PCR and western blot method respectively. RESULTS:27-OHC induced apoptosis in co-cultured SH-SY5Y cells and C6 cells. 27-OHC treatment significantly inhibited cell viability and proliferation (p<0.05). Compared with control group, 27-OHC caused the reduction of mitochondrial membrane potential (p<0.05). Additionally, the mRNA and protein expression of cholesterol synthesis-related factors, such as SREBP-1, HMGCR, were down-regulated (p<0.05), while the mRNA and protein expression of cholesterol transport-related factors (LXR-α, ABCA1) were up-regulated (p<0.05). CONCLUSIONS:Cytotoxicity and cholesterol metabolism disorder induced by 27-OHC may contribute to the pathogenesis of neurodegenerative disease.
Authors: Amanda J Unsworth; Alexander P Bye; Dionne S Tannetta; Michael J R Desborough; Neline Kriek; Tanya Sage; Harriet E Allan; Marilena Crescente; Parveen Yaqoob; Timothy D Warner; Chris I Jones; Jonathan M Gibbins Journal: Arterioscler Thromb Vasc Biol Date: 2017-06-15 Impact factor: 8.311