The synthesis of 25-Hydroxycholesterol is catalysed by cholesterol 25-hydroxylase, which uses cholesterol and molecular oxygen as substrates and NADPH as a cofactor. 25-Hydroxycholesterol is a potent biologically active lipid in the innate and adaptive immune system[1].
25-Hydroxycholesterol/25-羟基胆固醇 细胞实验
Cell Line
Concentration
Treated Time
Description
References
HEK293T cells
1µM
8h
Inhibited VSV-GFP expression
Immunity. 2013 Jan 24;38(1):92-105
Bovine aortic smooth muscle cells (BASMC)
1 μg/ml
24 days
25-Hydroxycholesterol did not induce nodule formation or calcification in BASMC.
J Clin Invest. 1994 May;93(5):2106-13
Bovine aortic endothelial cells (BAEC)
1 μg/ml
24 days
25-Hydroxycholesterol did not induce nodule formation or calcification in BAEC.
J Clin Invest. 1994 May;93(5):2106-13
Calcifying vascular cells (CVC)
1 μg/ml
24 days
25-Hydroxycholesterol significantly increased the number of mineralized nodules formed by CVC, which were highly calcified.
J Clin Invest. 1994 May;93(5):2106-13
PBMCs (Peripheral Blood Mononuclear Cells)
0.5 or 1.0 μM
12 h
To evaluate the inhibitory effect of 25-HC@DDAB on cytokine storm in PBMCs from COVID-19 patients. Results showed that 25-HC@DDAB significantly reduced levels of inflammatory cytokines such as IL-1β.
Nano Today. 2021 Jun;38:101149
BM-derived macrophages (BMDM)
25 ng/ml
4 h
To study the effect of 25-HC on lipid body formation, results showed that 25-HC induces lipid body formation.
J Exp Med. 2012 Apr 9;209(4):807-17
Caco2 cells
0.07 to 16.7 µM
20 h
Evaluate the inhibitory effect of 25HC and 27HC on HRV infection, results showed both significantly inhibited HRV infection.
Redox Biol. 2018 Oct;19:318-330
MA104 cells
0.07 to 16.7 µM
20 h
Evaluate the inhibitory effect of 25HC and 27HC on HRV infection, results showed both significantly inhibited HRV infection.
Redox Biol. 2018 Oct;19:318-330
splenic B220+ cells
250 nM
6 days
25-Hydroxycholesterol suppressed IgA class switch recombination in B cells, particularly when exposed early
Proc Natl Acad Sci U S A. 2009 Sep 29;106(39):16764-9
bone marrow-derived macrophages
500 nM
24 h
TLR4 activation led to significant increases in 25-hydroxycholesterol and CH25H mRNA and protein levels
Proc Natl Acad Sci U S A. 2009 Sep 29;106(39):16764-9
apoE4-expressing microglia
10 μg/ml
24 h
apoE4-expressing microglia secrete higher levels of IL-1β after treatment with LPS and 25-HC
J Neuroinflammation. 2020 Jun 17;17(1):192
Primary mouse microglia (PMG)
5 μg/ml
24 h
To investigate the effect of 25HC on microglial phagocytic capacity, results showed that 25HC significantly reduced the phagocytic ability of microglia.
J Neuroinflammation. 2025 Feb 25;22(1):50
CEM cells
5µM
12h
Inhibited HIV entry
Immunity. 2013 Jan 24;38(1):92-105
HeLa cells
1µM
5h
Inhibited Ebola virus replication
Immunity. 2013 Jan 24;38(1):92-105
PBMCs
1µM
24h
Inhibited HIV NL4-3 p24 expression
Immunity. 2013 Jan 24;38(1):92-105
J2 BMMs
1µM
12h
Reduced VSV replication
Immunity. 2013 Jan 24;38(1):92-105
Bone marrow-derived macrophages (BMDMs)
1 μM
24 h
To evaluate the effect of 25-HC on IL-1β production after L. monocytogenes infection. Results showed that Ch25h−/− BMDMs produced more IL-1β, and this effect was inhibited by the addition of 25-HC.
Cell. 2017 Nov 16;171(5):1057-1071. e11
25-Hydroxycholesterol/25-羟基胆固醇 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
C57BL/6 mice
Cecal ligation and puncture (CLP)-induced sepsis model
Intravenous injection
1 μM
Single dose, observed for 96 hours
To evaluate the therapeutic effect of 25-HC@DDAB in a CLP-induced sepsis mouse model. Results showed that 25-HC@DDAB significantly improved survival rate (50%) and reduced inflammation and tissue damage.
Nano Today. 2021 Jun;38:101149
Mouse
5XFAD transgenic mice
Intraperitoneal injection
10 mg/kg
Every other day for four weeks
To investigate the effect of 25HC on Alzheimer's disease pathology, results showed that 25HC accelerated amyloid accumulation and exacerbated cognitive and behavioral deficits.
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