Cholesterol sulfate is a naturally occurring, orally active cholesterol derivative that is widely distributed in various tissues and body fluids. Cholesterol sulfate acts as a DOCK2 inhibitor, with IC50 values of 2 μM and 2.9 μM against mouse and human targets, respectively. Cholesterol sulfate restricts excessive neutrophil infiltration and alleviates intestinal inflammation and damage. Cholesterol sulfate serves as an activator of protein kinase C ( PKC ), which promotes squamous cell differentiation and inhibits skin carcinogenesis. Cholesterol sulfate regulates cholesterol homeostasis and cellular metabolism by activating the AMPK-Sirt1 pathway. Cholesterol sulfate can be used in research related to actinic keratitis, ulcerative colitis, skin cancer, and other conditions [1] [2] [3] [4] [5] [6].
In Vivo, Cholesterol sulfate (8 μg/μL; topical; 6 total doses) suppresses inflammatory cell infiltration in the anterior chamber of UV-induced photokeratitis models in Sult2b1−/− mice[1]. Cholesterol sulfate (8 μg/μL; topical; three times daily; for 3 days) suppresses inflammatory cell infiltration in the conjunctiva of experimental allergic conjunctivitis models in Sult2b1−/− mice[1]. Cholesterol sulfate (200 mg/kg; p.o.; three times at 4-hour intervals) ameliorates Indomethacin -induced small intestinal ulceration and reduces neutrophil infiltration into ulcerative lesions in Sult2b1-/- mice[2]. Cholesterol sulfate (0.004%; dietary supplementation; continued for 6 days) alleviates 2.5% DSS-induced acute ulcerative colitis in Sult2b1ΔIEC mice by reducing disease severity markers and promoting colonic epithelial cell proliferation[3]. Cholesterol sulfate (20 mg/kg; subcutaneous (calvarial); 2 doses (Days 0 and 2)) inhibits LPS-induced bone destruction and osteoclast formation in male C57BL/6 mice, reducing bone cavity formation by ~6-fold and TRAP-positive osteoclasts by ~24-fold[5]. Cholesterol sulfate (20 mg/kg; intraperitoneal; six times weekly; 3 weeks) protects against ovariectomy-induced bone loss in female C57BL/6 mice, restoring key bone density parameters and increasing osteoclast apoptosis to ~55% of TRAP-positive cells[5]. Cholesterol sulfate (400 µg; topical; weekly; 19 weeks) inhibits skin tumor promotion in mice, reducing tumor incidence by 56%, tumor number per mouse by 81%, and tumor size by 60% at 20 weeks[6].
In Vitro, Cholesterol sulfate (0.19-150 μM; 20 min-6 h) inhibits chemokine-induced Rac activation in mouse T cells and bone marrow-derived neutrophils, thereby blocking the migration of these cells in vitro[1]. Cholesterol sulfate (12.5 μM; 60 min) potently inhibits phorbol ester (PMA) -induced reactive oxygen species (ROS) production in mouse bone marrow-derived neutrophils[2]. Cholesterol sulfate is a sulfated cholesterol. At concentrations of 5-100 μM for 24 h, it activates the SREBP2 signaling pathway in a dose-dependent manner in SULT2B1-knockout human colon cancer cell line HT-29, and this activating effect is enhanced upon SLC10A6 overexpression[3]. Cholesterol sulfate (25-50 μM; 6-18 h) promotes the proteolytic activation of SREBP2 in HEK293T cells[3]. Cholesterol sulfate (50 μM; 24 h) upregulates the expression of key cholesterol biosynthesis genes in human colonic epithelial cell lines HT-29, LOVO, SW480, HCT116, SW1116 and NCM460[3]. Cholesterol sulfate (50 μM; 6-48 h) increases the levels of total cholesterol and free cholesterol in HT-29 cells and SULT2B1-knockout HT-29 human colon cancer cells[3]. Cholesterol sulfate (25-50 μM) increases the cell viability of HT-29 human colon cancer cells treated with 25 or 50 μM cholesterol sulfate, as well as SULT2B1-knockout HT-29 human colon cancer cells, and this effect depends on the activation of SREBP2[3]. Cholesterol sulfate (50 μM) increases the level of mature nuclear-localized SREBP2 in HT-29 cells and SULT2B1-knockout HT-29 human colon cancer cells[3]. Cholesterol sulfate (25 μM; 12-48 h) significantly reduces the intracellular total cholesterol level by 20% to 30% in HEK 293T, Huh-7 and MEF cells[4]. Cholesterol sulfate (50 μM; 24 h) significantly reduces intracellular cholesterol levels in Huh-7 cells[4]. Cholesterol sulfate (25 μM; 12 h) inhibits de novo cholesterol synthesis in HEK 293T, Huh-7 and MEF cells[4]. Cholesterol sulfate (25 μM) can partially inhibit LDL-cholesterol uptake by cholesterol-depleted Huh-7 cells switched to FBS-containing medium[4]. Cholesterol sulfate (3.12-25 μM; 5-16 h) reduces the protein expression level of HMGCR in Huh-7 cells at the post-translational level, with an IC50 of 5.6 μM for inhibiting T7-tagged HMGCR after 5 h of treatment[4]. Cholesterol sulfate (12.5-25 μM; 5 h) promotes ubiquitination and proteasomal degradation of wild-type HMGCR in Huh-7 cells, and this process depends on Lys89 and Lys248 of HMGCR[4]. Cholesterol sulfate (25 μM; 8 h) promotes the interaction between INSIG1 and HMGCR in HEK 293T cells, a process that mediates the ubiquitination and degradation of HMGCR[4]. Cholesterol sulfate (25 μM; 8 h) promotes the interaction between INSIG1 and SCAP in HEK 293T cells, thereby inhibiting the translocation of SREBP2 to the Golgi apparatus; this interaction depends on the L343 and V355 residues of SCAP, but not on I348[4]. Cholesterol sulfate (25 μM; 4-24 h) reduces LDL-cholesterol uptake in Huh-7 cells through two mechanisms: a secondary effect of cholesterol depletion, and direct inhibition of clathrin-mediated endocytosis[4]. Cholesterol sulfate (25 μM; 4-24 h) inhibits LDLR endocytosis in Huh-7 cells, resulting in the accumulation of LDLR on the cell surface[4]. Cholesterol sulfate (25 μM; 8 h) partially inhibits the upregulation of SREBP2 target genes in cholesterol-depleted Huh-7 cells[4]. Cholesterol sulfate (25 μM) inhibits the proteolytic processing of SREBP2 in cholesterol-depleted Huh-7 cells and reduces the level of the active nuclear form of SREBP2[4]. Cholesterol sulfate (20-40 μM; 2-4 days) inhibits RANKL-induced osteoclast differentiation and NFATc1 pathway activation in mouse bone marrow macrophages (BMMs) without reducing cell viability; treatment with 30 μM for 2 days suppresses the expression of NFATc1 and its target genes, and this inhibitory effect persists for up to 4 days[5]. Cholesterol sulfate (30 μM; 2-4 days) inhibits RANKL-induced osteoclast differentiation in a RORα-independent manner, as it exerts comparable inhibitory effects on osteoclast formation and NFATc1 expression in bone marrow macrophages (BMMs) from wild-type (WT) and RORα-deficient mice[5]. Cholesterol sulfate (30 μM; 0.5-24 h) activates the AMPK-Sirt1 axis in a RORα-independent manner, thereby inhibiting NF-κB activity in mouse bone marrow-derived macrophages (BMMs) and RAW264.7 cells[5]. Cholesterol sulfate (30 μM; 10 h-8 days) induces caspase-dependent apoptosis in differentiated murine osteoclasts, disrupts their actin ring structure, and inhibits bone resorptive activity[5]. Cholesterol sulfate (30 μM; 12 h-4 days) induces apoptosis of mouse osteoclasts via AMPK-dependent NF-κB inhibition, thereby reducing the production of IL-1β; treatment with IL-1β reverses the aforementioned pro-apoptotic and inhibitory effects without altering the activation level of AMPK[5]. Cholesterol sulfate induces squamous differentiation of normal human keratinocytes by inhibiting cell growth and upregulating the expression and activity of TGase 1[6]. Cholesterol sulfate induces granular cell differentiation in mouse basal keratinocytes and regulates the expression of differentiation markers[6].
References:
[1]. Sakurai T, et al. Cholesterol sulfate is a DOCK2 inhibitor that mediates tissue-specific immune evasion in the eye. Sci Signal. 2018;11(541):eaao4874. Published 2018 Jul 31.
[2]. Morino K, et al. Cholesterol sulfate limits neutrophil recruitment and gut inflammation during mucosal injury. Front Immunol. 2023;14:1131146. Published 2023 Mar 17.
[3]. Xu D, et al. Cholesterol sulfate alleviates ulcerative colitis by promoting cholesterol biosynthesis in colonic epithelial cells. Nature Communications. 2022 Jul 30;13(1):4428.
[4]. Nam LB, et al. Cholesterol sulfate as a negative regulator of cellular cholesterol homeostasis. Mol Cells. 2025;48(6):100209.
[5]. Park JH, et al. Cholesterol sulfate inhibits osteoclast differentiation and survival by regulating the AMPK-Sirt1-NF-κB pathway. J Cell Physiol. 2023;238(9):2063-2075.
[6]. Kuroki T, et al. Cholesterol sulfate, an activator of protein kinase C mediating squamous cell differentiation: a review. Mutat Res. 2000;462(2-3):189-195.
In Vivo, Cholesterol sulfate (8 μg/μL; topical; 6 total doses) suppresses inflammatory cell infiltration in the anterior chamber of UV-induced photokeratitis models in Sult2b1−/− mice[1]. Cholesterol sulfate (8 μg/μL; topical; three times daily; for 3 days) suppresses inflammatory cell infiltration in the conjunctiva of experimental allergic conjunctivitis models in Sult2b1−/− mice[1]. Cholesterol sulfate (200 mg/kg; p.o.; three times at 4-hour intervals) ameliorates Indomethacin -induced small intestinal ulceration and reduces neutrophil infiltration into ulcerative lesions in Sult2b1-/- mice[2]. Cholesterol sulfate (0.004%; dietary supplementation; continued for 6 days) alleviates 2.5% DSS-induced acute ulcerative colitis in Sult2b1ΔIEC mice by reducing disease severity markers and promoting colonic epithelial cell proliferation[3]. Cholesterol sulfate (20 mg/kg; subcutaneous (calvarial); 2 doses (Days 0 and 2)) inhibits LPS-induced bone destruction and osteoclast formation in male C57BL/6 mice, reducing bone cavity formation by ~6-fold and TRAP-positive osteoclasts by ~24-fold[5]. Cholesterol sulfate (20 mg/kg; intraperitoneal; six times weekly; 3 weeks) protects against ovariectomy-induced bone loss in female C57BL/6 mice, restoring key bone density parameters and increasing osteoclast apoptosis to ~55% of TRAP-positive cells[5]. Cholesterol sulfate (400 µg; topical; weekly; 19 weeks) inhibits skin tumor promotion in mice, reducing tumor incidence by 56%, tumor number per mouse by 81%, and tumor size by 60% at 20 weeks[6].
In Vitro, Cholesterol sulfate (0.19-150 μM; 20 min-6 h) inhibits chemokine-induced Rac activation in mouse T cells and bone marrow-derived neutrophils, thereby blocking the migration of these cells in vitro[1]. Cholesterol sulfate (12.5 μM; 60 min) potently inhibits phorbol ester (PMA) -induced reactive oxygen species (ROS) production in mouse bone marrow-derived neutrophils[2]. Cholesterol sulfate is a sulfated cholesterol. At concentrations of 5-100 μM for 24 h, it activates the SREBP2 signaling pathway in a dose-dependent manner in SULT2B1-knockout human colon cancer cell line HT-29, and this activating effect is enhanced upon SLC10A6 overexpression[3]. Cholesterol sulfate (25-50 μM; 6-18 h) promotes the proteolytic activation of SREBP2 in HEK293T cells[3]. Cholesterol sulfate (50 μM; 24 h) upregulates the expression of key cholesterol biosynthesis genes in human colonic epithelial cell lines HT-29, LOVO, SW480, HCT116, SW1116 and NCM460[3]. Cholesterol sulfate (50 μM; 6-48 h) increases the levels of total cholesterol and free cholesterol in HT-29 cells and SULT2B1-knockout HT-29 human colon cancer cells[3]. Cholesterol sulfate (25-50 μM) increases the cell viability of HT-29 human colon cancer cells treated with 25 or 50 μM cholesterol sulfate, as well as SULT2B1-knockout HT-29 human colon cancer cells, and this effect depends on the activation of SREBP2[3]. Cholesterol sulfate (50 μM) increases the level of mature nuclear-localized SREBP2 in HT-29 cells and SULT2B1-knockout HT-29 human colon cancer cells[3]. Cholesterol sulfate (25 μM; 12-48 h) significantly reduces the intracellular total cholesterol level by 20% to 30% in HEK 293T, Huh-7 and MEF cells[4]. Cholesterol sulfate (50 μM; 24 h) significantly reduces intracellular cholesterol levels in Huh-7 cells[4]. Cholesterol sulfate (25 μM; 12 h) inhibits de novo cholesterol synthesis in HEK 293T, Huh-7 and MEF cells[4]. Cholesterol sulfate (25 μM) can partially inhibit LDL-cholesterol uptake by cholesterol-depleted Huh-7 cells switched to FBS-containing medium[4]. Cholesterol sulfate (3.12-25 μM; 5-16 h) reduces the protein expression level of HMGCR in Huh-7 cells at the post-translational level, with an IC50 of 5.6 μM for inhibiting T7-tagged HMGCR after 5 h of treatment[4]. Cholesterol sulfate (12.5-25 μM; 5 h) promotes ubiquitination and proteasomal degradation of wild-type HMGCR in Huh-7 cells, and this process depends on Lys89 and Lys248 of HMGCR[4]. Cholesterol sulfate (25 μM; 8 h) promotes the interaction between INSIG1 and HMGCR in HEK 293T cells, a process that mediates the ubiquitination and degradation of HMGCR[4]. Cholesterol sulfate (25 μM; 8 h) promotes the interaction between INSIG1 and SCAP in HEK 293T cells, thereby inhibiting the translocation of SREBP2 to the Golgi apparatus; this interaction depends on the L343 and V355 residues of SCAP, but not on I348[4]. Cholesterol sulfate (25 μM; 4-24 h) reduces LDL-cholesterol uptake in Huh-7 cells through two mechanisms: a secondary effect of cholesterol depletion, and direct inhibition of clathrin-mediated endocytosis[4]. Cholesterol sulfate (25 μM; 4-24 h) inhibits LDLR endocytosis in Huh-7 cells, resulting in the accumulation of LDLR on the cell surface[4]. Cholesterol sulfate (25 μM; 8 h) partially inhibits the upregulation of SREBP2 target genes in cholesterol-depleted Huh-7 cells[4]. Cholesterol sulfate (25 μM) inhibits the proteolytic processing of SREBP2 in cholesterol-depleted Huh-7 cells and reduces the level of the active nuclear form of SREBP2[4]. Cholesterol sulfate (20-40 μM; 2-4 days) inhibits RANKL-induced osteoclast differentiation and NFATc1 pathway activation in mouse bone marrow macrophages (BMMs) without reducing cell viability; treatment with 30 μM for 2 days suppresses the expression of NFATc1 and its target genes, and this inhibitory effect persists for up to 4 days[5]. Cholesterol sulfate (30 μM; 2-4 days) inhibits RANKL-induced osteoclast differentiation in a RORα-independent manner, as it exerts comparable inhibitory effects on osteoclast formation and NFATc1 expression in bone marrow macrophages (BMMs) from wild-type (WT) and RORα-deficient mice[5]. Cholesterol sulfate (30 μM; 0.5-24 h) activates the AMPK-Sirt1 axis in a RORα-independent manner, thereby inhibiting NF-κB activity in mouse bone marrow-derived macrophages (BMMs) and RAW264.7 cells[5]. Cholesterol sulfate (30 μM; 10 h-8 days) induces caspase-dependent apoptosis in differentiated murine osteoclasts, disrupts their actin ring structure, and inhibits bone resorptive activity[5]. Cholesterol sulfate (30 μM; 12 h-4 days) induces apoptosis of mouse osteoclasts via AMPK-dependent NF-κB inhibition, thereby reducing the production of IL-1β; treatment with IL-1β reverses the aforementioned pro-apoptotic and inhibitory effects without altering the activation level of AMPK[5]. Cholesterol sulfate induces squamous differentiation of normal human keratinocytes by inhibiting cell growth and upregulating the expression and activity of TGase 1[6]. Cholesterol sulfate induces granular cell differentiation in mouse basal keratinocytes and regulates the expression of differentiation markers[6].
References:
[1]. Sakurai T, et al. Cholesterol sulfate is a DOCK2 inhibitor that mediates tissue-specific immune evasion in the eye. Sci Signal. 2018;11(541):eaao4874. Published 2018 Jul 31.
[2]. Morino K, et al. Cholesterol sulfate limits neutrophil recruitment and gut inflammation during mucosal injury. Front Immunol. 2023;14:1131146. Published 2023 Mar 17.
[3]. Xu D, et al. Cholesterol sulfate alleviates ulcerative colitis by promoting cholesterol biosynthesis in colonic epithelial cells. Nature Communications. 2022 Jul 30;13(1):4428.
[4]. Nam LB, et al. Cholesterol sulfate as a negative regulator of cellular cholesterol homeostasis. Mol Cells. 2025;48(6):100209.
[5]. Park JH, et al. Cholesterol sulfate inhibits osteoclast differentiation and survival by regulating the AMPK-Sirt1-NF-κB pathway. J Cell Physiol. 2023;238(9):2063-2075.
[6]. Kuroki T, et al. Cholesterol sulfate, an activator of protein kinase C mediating squamous cell differentiation: a review. Mutat Res. 2000;462(2-3):189-195.