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  4. Cholesterol

Cholesterol is the major sterol in mammals. It is making up 20-25% of structural component of the plasma membrane. Plasma membranes are highly permeable to water but relatively impermeable to ions and protons. Cholesterol plays an important role in determining the fluidity and permeability characteristics of the membrane as well as the function of both the transporters and signaling proteins. Cholesterol is also an endogenous estrogen-related receptor α (ERRα) agonist.

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Cholesterol Chemical Structure

Cholesterol Chemical Structure

CAS No. : 57-88-5

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Based on 19 publication(s) in Google Scholar

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Description

Cholesterol is the major sterol in mammals. It is making up 20-25% of structural component of the plasma membrane. Plasma membranes are highly permeable to water but relatively impermeable to ions and protons. Cholesterol plays an important role in determining the fluidity and permeability characteristics of the membrane as well as the function of both the transporters and signaling proteins[1][2]. Cholesterol is also an endogenous estrogen-related receptor α (ERRα) agonist[3].

IC50 & Target

Microbial Metabolite

 

Human Endogenous Metabolite

 

In Vitro

GT1-7 hypothalamic cells subjected to Cholesterol depletion in vitro produced 20-31% reductions in cellular Cholesterol content. All Cholesterol-depleted neuron-derived cells, exhibit decreased phosphorylation/activation of IRS-1 and AKT following stimulation by insulin, insulin-like growth factor-1, or the neurotrophins (NGF and BDNF). Reduction in cellular Cholesterol also results in increased basal autophagy and impairment of induction of autophagy by glucose deprivation[1].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

Cholesterol can be used in animal modeling to construct a rat model of hyperlipidemia.

Insulin-deficient diabetes in mice can lead to a reduction in brain Cholesterol synthesis, which occurs through a change in expression of Cholesterol synthesis enzymes and their upstream regulators SREBP2 and SCAP1[1].
The pool of Cholesterol in the whole animal is 2,200 mg/kg body weight. This is true for essentially all species from the mouse to the primate and indicates that the average concentration of Cholesterol in the whole animal is 2.2 mg/g fresh tissue[2].
The basal metabolic rate in the mouse is 170 kilocalories (kcal)/day/kg, and the flow of Cholesterol from all peripheral organs to the liver is greater than 100 mg/day/kg[2].The metabolic half-life of Cholesterol varies with the type of lipoprotein it binds to and the different tissues it is located in, ranging from a few hours to several years. Cholesterol can be used for modeling purposes related to hyperlipidemia and atherosclerosis[4].

Induction of Hyperlipidemia[5][6]
Background
Hyperlipidemia is a group of disorders characterized by elevated concentrations of circulating lipids, including cholesterol, cholesterol esters, phospholipids and triglycerides. If the intake of cholesterol is too much, and exceeds the body's metabolic capacity, it may lead to increased plasma cholesterol levels, causing hyperlipidemia.
Specific Mmodeling Methods
Rats: Wistar • male • 18-week-old (period: 8 weeks)
Administration: 2% cholesterol; diet • 8 weeks
Note
(1) Rats were housed in a room maintained at a 12-h light-dark cycle and a constant temperature of 22±2 °C
(2) Wistar rats were always chosen for hyperlipidemia studies since this species shows a moderate increase in serum cholesterol and triglyceride level due to a high-cholesterol diet and no substantial atherosclerosis develops; therefore, the direct myocardial effect of hyperlipidemia, independent from atherosclerosis, can be studied in this model.
Modeling Indicators
Molecular changes: Significant increase in total cholesterol levels in blood samples (about 20%)
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Induction of atherosclerosis[7][8]
Background
High levels of cholesterol in the blood, especially low-density lipoprotein cholesterol (LDL-C), can accumulate plaque on the walls of blood vessels, a process known as atherosclerosis. Over time, these plaques can block blood flow and cause serious health problems such as myocardial ischemia or myocardial infarction.
Specific Mmodeling Methods
Rabbits: Oryctolagus cuniculus • male • 4–6-month-old (period: 16 weeks)
Administration: 0.3% cholesterol and 3% soybean oil; diet • 16 weeks
Note
(1) The cholesterol-fed rabbit is a widely used model for experimental atherosclerosis research as cholesterol only cause atherosclerotic changes in the rabbit arterial intima, which was very similar to human atherosclerosis.
(2) As the absorption of dietary cholesterol requires fat, you must add oil into the diet. Otherwise, rabbits will use their internal fat, which makes them lean or sick. In addition, using soybean oil, which consists of unsaturated fatty acids, can prevent the levels of plasma cholesterol from becoming too high. Other vegetable oils, such as peanut oil or corn oil, can be used because they are all unsaturated fatty acids. Animal fat (saturated fatty acids) like tallow and lard is not recommended.
(3) 0.3–0.5% cholesterol diet is recommended for most experiments. Rabbits cannot tolerate a 1–2% cholesterol diet for a month as they develop severe liver dysfunction.
(4) Adult rabbits at 4 months or older can consume approximately ~150 g a day. You can either feed ab libitum or restricted (100–150 g/day/adult rabbit).
(5) Plasma lipids should be measured weekly, especially for the first 4 weeks, because you need to determine whether plasma levels of cholesterol are elevated in each animal. Non-responder rabbits can be excluded from the experiments if their plasma cholesterol levels do not increase after cholesterol diet feeding.
(6) Plasma lipoproteins can be measured at 8 and 16 weeks when the plasma levels of cholesterol are stable.
(7) The age of rabbits should be considered because young rabbits are more susceptible to aortic atherosclerosis than old rabbits even though they have similar plasma cholesterol levels. 4–6-month-old rabbits are usually used for cholesterol feeding experiments.
(8) Male and female rabbits are different in terms of response to a cholesterol diet and atherosclerosis. In our experience, female rabbits develop higher hypercholesterolemia and greater aortic lesions than their counterpart male rabbits. In general, male rabbits are recommended for experiments because estrogen may influence the results.
Modeling Indicators
Histological changes: atherosclerosis lesions can be seen on HE stained aortic arch and thoracic aorta segments
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MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Clinical Trial
Molecular Weight

386.65

Formula

C27H46O

CAS No.
Appearance

Solid

Color

White to off-white

SMILES

O[C@H](C1)CC[C@@]2(C)C1=CC[C@]3([H])[C@]2([H])CC[C@@]4(C)[C@@]3([H])CC[C@]4([H])[C@@H](CCCC(C)C)C

Structure Classification
Initial Source
Shipping

Room temperature in continental US; may vary elsewhere.

Storage
Powder -20°C 3 years
4°C 2 years
In solvent -80°C 6 months
-20°C 1 month
Solvent & Solubility
In Vitro: 

Ethanol : 20 mg/mL (51.73 mM; Need ultrasonic)

DMSO : < 1 mg/mL (insoluble or slightly soluble)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.5863 mL 12.9316 mL 25.8632 mL
5 mM 0.5173 mL 2.5863 mL 5.1726 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

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Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

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In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% EtOH    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 1.43 mg/mL (3.70 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.43 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (14.3 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% EtOH    90% Corn Oil

    Solubility: ≥ 1.43 mg/mL (3.70 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.43 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.

    Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (14.3 mg/mL) to 900 μL Corn oil, and mix evenly.

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
Purity & Documentation

Purity: 99.94%

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
Ethanol 1 mM 2.5863 mL 12.9316 mL 25.8632 mL 64.6580 mL
5 mM 0.5173 mL 2.5863 mL 5.1726 mL 12.9316 mL
10 mM 0.2586 mL 1.2932 mL 2.5863 mL 6.4658 mL
15 mM 0.1724 mL 0.8621 mL 1.7242 mL 4.3105 mL
20 mM 0.1293 mL 0.6466 mL 1.2932 mL 3.2329 mL
25 mM 0.1035 mL 0.5173 mL 1.0345 mL 2.5863 mL
30 mM 0.0862 mL 0.4311 mL 0.8621 mL 2.1553 mL
40 mM 0.0647 mL 0.3233 mL 0.6466 mL 1.6164 mL
50 mM 0.0517 mL 0.2586 mL 0.5173 mL 1.2932 mL
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Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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Cholesterol
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