Lovastatin

Manufactured by Enzo Life Sciences

Sourced in United States

Lovastatin is a laboratory reagent used for research purposes. It is a fungal-derived compound that acts as an inhibitor of the enzyme HMG-CoA reductase, a key enzyme involved in the biosynthesis of cholesterol. Lovastatin is commonly used as a research tool to study cholesterol metabolism and related biochemical processes.

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5 protocols using lovastatin

Quantification of Cellular Cholesterol

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For determination of cellular cholesterol levels, human GFs were treated with IL1β, TNFα, cholesterol synthesis inhibitors (triparanol (Cayman) and lovastatin (Enzo), or anti-CXCL16 blocking antibody (R&D Systems) in the presence of IL1β (2 ng ml⁻¹), TNFα (50 ng ml⁻¹) or cholesterol (200 μM). The quantity of intracellular cholesterol was determined by a cholesterol quantification kit (Biomax, BM-CHO−100). Briefly, cellular lipids were extracted with a mixture of chloroform, isopropanol, and NP-40 (7:11:0.1) in a microhomogenizer and centrifuged for 10 min at 15,000 × g. The organic phase was collected and dried under vacuum at 55 °C for 12 h. From the lipid extract obtained, the amounts of total and free cholesterol were determined using a total cholesterol quantification kit following the manufacturer’s instructions (Biomax). The cholesteryl ester content was calculated as the total cholesterol minus free cholesterol.

Tran T.T., Lee G., Huh Y.H., Chung K.H., Lee S.Y., Park K.H., Kwon S.H., Kook M.S., Chun J.S., Koh J.T, & Ryu J.H. (2023). Disruption of cholesterol homeostasis triggers periodontal inflammation and alveolar bone loss. Experimental & Molecular Medicine, 55(12), 2553-2563.

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Compound Validation in Cell Culture

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Lovastatin (Enzo Life Sciences), Tranylcypromine (Enzo Life Sciences), Chloroquine Phosphate (CQ, Enzo Life Sciences), Acetylsalicylic Acid (AA, Sigma-Aldrich), Sulindac (Enzo Life Sciences), Mycophenolate Mofetil (MMF, Enzo Life Sciences), and Amifostine (Enzo Life Sciences) were prepared for validation as a stock solution in DMSO or H₂O and dilutions were produced to obtain working concentrations in cell culture medium.

Zanni E.D., Bianchi G., Ravazzolo R., Raffaghello L., Ceccherini I, & Bachetti T. (2017). Targeting of PHOX2B expression allows the identification of drugs effective in counteracting neuroblastoma cell growth. Oncotarget, 8(42), 72133-72146.

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Statin Drugs and Cholesterol Synthesis

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Nine statin drugs were obtained from commercial sources: Atorvastatin calcium salt trihydrate, simvastatin, pravastatin, mevastatin (PZ0001, S6196, P4498, M2537 Sigma, St Louis, MO, USA); fluvastatin, rosuvastatin, cerivastatin (F601250, R700500, C277000, Toronto Research Chemicals Inc., Toronto, ON, Canada); pitavastatin (S1759, Selleckchem, Boston, MA, USA); lovastatin (430-103-M050, Enzo life sciences, Farmingdale, NY, USA). We prepared 10 mM stock solution in DMSO for in vitro testing. For in vivo testing, pitavastatin was prepared as fresh suspension in methycellulose at 0.4 mg ml⁻¹ (for oral gavage). fluvastatin, cerivastatin, and pitavastatin were prepared as a solution at 0.25 mg ml⁻¹ in PBS for IP injection. GGTI-298 and FTI-277 (G5169, A1393, Sigma) were made into 10 mM stock solution in DMSO. AICAR (#S1802, Selleckchem) was made into 100 mM stock solution in PBS. Intermediate products of cholesterol synthesis were purchased from Sigma: mevalonolactone (M4667), farnesyl pyrophosphate ammonium salt (F6892), geranylgeranyl pyrophosphate ammonium salt (G6025), squalene (S3626), cholesterol solution (S5442), geranylpyrophosphate (G6772), and isopentenyl pyrophosphate triammonium salt solution (I0503).

Jiang P., Mukthavaram R., Chao Y., Nomura N., Bharati I.S., Fogal V., Pastorino S., Teng D., Cong X., Pingle S.C., Kapoor S., Shetty K., Aggrawal A., Vali S., Abbasi T., Chien S, & Kesari S. (2014). In vitro and in vivo anticancer effects of mevalonate pathway modulation on human cancer cells. British Journal of Cancer, 111(8), 1562-1571.

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DiI-Labeled Oxidized LDL Binding Assay

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Filipin, MβCD, glimepiride and atorvastatin were purchased from Sigma-Aldrich (St. Louis, MO). Lovastatin (Enzo Life Sciences Inc., Farmingdale, NY) was activated by the addition of 0,1 N NaOH (Lovastatin/NaOH 2:3 v/v), at 50°C and neutralized with 1 N HCl to pH 7,2.
Phenylmethylsulfonylfuoride (PMSF) (Euroclone, Devon, UK), pepstatin A (Boehringer, Mannheim, DE), GM6001 inhibitor (Biomol Research Lab Inc.), protease inhibitor cocktail set III (AEBSF 100mM; aprotinin 80μM; bestatin 5mM; E-64 1,5mM; leupeptin 2mM; pepstatin A 1mM) (Calbiochem, La Jolla, CA) were used for specific protease class inhibition experiments.
Ox-LDL preparation, labelling and fluorescence analysis
Human LDL was prepared from fresh healthy normolipidemic plasma of volunteers by ultracentrifugation [50 ] and in vitro oxidized as described (Matarazzo et al., 2012). Ox-LDL was labelled with 1,1-dioctadecyl-3,3,3,3-tetramethyllindocarbocyanine perchlorate (DiI, Invitrogen, Paisley, UK) [7 (link),51 (link)]. DiI-ox-LDL binding assay was performed as previously described [13 (link)].

Gioia M., Vindigni G., Testa B., Raniolo S., Fasciglione G.F., Coletta M, & Biocca S. (2015). Membrane Cholesterol Modulates LOX-1 Shedding in Endothelial Cells. PLoS ONE, 10(10), e0141270.

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Statin regulation of T-cell activation

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Atorvastatin, Fluvastatin, Lovastatin, Pravastatin, Rosuvastatin and Simvastatin were purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA); Pitavastatin, cholesterol and U0126 from Selleckchem (Houston, USA); Farnesyl Pyrophosphate (FPP) and Geranylgeranyl Pyrophosphate (GGPP) from Biomol GmbH (Hamburg, Germany); and Mevalonic acid lactone (MVA) from Sigma Aldrich (St. Louis, MO, USA). Hybridoma supernatants containing mouse anti-human CD3ε (OKT-3) and CD28 (248.23.2) monoclonal antibodies were produced and purified in our laboratory.

Voss L., Guttek K., Reddig A., Reinhold A., Voss M., Simeoni L., Schraven B, & Reinhold D. (2021). Pitavastatin Is a Highly Potent Inhibitor of T-Cell Proliferation. Pharmaceuticals, 14(8), 727.

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