“…Statin therapy reduces the synthesis of cholesterol and increases its absorption. These findings suggest that a change in one vector, consistently, results in a compensatory and opposing change in the other [29].…”
Section: Physiological Factors Affecting Cholesterol Absorption In Inmentioning
The goal of this review is to discuss many aspects of cholesterol homeostasis in the body in order to presents the standards of cholesterol disposition and its levels, especially when several means might change the features of cholesterol synthesis and absorption and interrupt this equilibrium. With the detection of pathways of cholesterol absorption and the discovery of different compounds that amend cholesterol absorption, there is a series of cholesterol lowering drugs, which may decrease the level of serum cholesterol by altering its absorption. It is recognized that the effects of cholesterol lowering drugs as well as genetics of a person, loss of body weight, enzymes and proteins may increase the cholesterol altering effect, but the impact of these agents on the mechanism of cholesterol homeostasis are poorly understood. The disturbance of cholesterol homeostasis is also related with vascular changes associated with cardiac and kidney problems. Besides using different drugs and other means (plant sterol, enzyme, protein etc.), there is a need to improve the dietary pattern and life style. These may help to improve the cholesterol homeostasis.
“…Statin therapy reduces the synthesis of cholesterol and increases its absorption. These findings suggest that a change in one vector, consistently, results in a compensatory and opposing change in the other [29].…”
Section: Physiological Factors Affecting Cholesterol Absorption In Inmentioning
The goal of this review is to discuss many aspects of cholesterol homeostasis in the body in order to presents the standards of cholesterol disposition and its levels, especially when several means might change the features of cholesterol synthesis and absorption and interrupt this equilibrium. With the detection of pathways of cholesterol absorption and the discovery of different compounds that amend cholesterol absorption, there is a series of cholesterol lowering drugs, which may decrease the level of serum cholesterol by altering its absorption. It is recognized that the effects of cholesterol lowering drugs as well as genetics of a person, loss of body weight, enzymes and proteins may increase the cholesterol altering effect, but the impact of these agents on the mechanism of cholesterol homeostasis are poorly understood. The disturbance of cholesterol homeostasis is also related with vascular changes associated with cardiac and kidney problems. Besides using different drugs and other means (plant sterol, enzyme, protein etc.), there is a need to improve the dietary pattern and life style. These may help to improve the cholesterol homeostasis.
“…Although the exact reason remains unknown, sitosterol is most effectively returned to the gut among all plant sterols, thereby resulting in the lowest absorption rate ( 38 ). Another possible explanation for these results is that orlistat limits dietary cholesterol absorption by the inhibition of Niemann-Pick C1-like 1 (NPC1L1) transport protein, as well as by the inhibition of intestinal lipase ( 39 ). The NPC1L1 was proposed to play an important role in the competitive uptake of plantar sterols and cholesterol across the enterocytes’ apical membrane ( 40 ).…”
BackgroundOrlistat, a reversible inhibitor of pancreatic and gastric lipase, is known to have anti-obesity and antioxidant properties. Cholesterol intermediates and metabolites have diverse and important functions in cardiovascular disease. Therefore, we aimed to evaluate the effect of orlistat on sterol metabolism in overweight and obese adults after weight loss during the intervention or weight loss at 12 weeks.MethodsA total of 51 (27 in the control group and 24 in the experimental group), patients with a BMI of 27 or greater were randomly assigned in a 1:1 ratio to receive either orlistat (120 mg) three times a day plus phentermine hydrochloride (37.5 mg) once daily or a placebo three times a day plus phentermine hydrochloride (37.5 mg) once daily. The primary study outcome was sterol metabolism.ResultsThe experimental group exhibited significantly decreased metabolic signatures of serum sterols, free cholesterol, sitosterol, 7α-hydroxycholesterol (7α-OHC), and 7β-OHC at 12 weeks. The experimental group also exhibited significantly decreased metabolic ratios of sitosterol and 7α-OHC to cholesterol at 12 weeks. Regarding changes in sterol signatures from baseline to 6-month follow-up, free cholesterol, plant sterols, and cholesterol precursors tended to decrease with weight loss during the intervention and increase again as the weight was regained in both groups.ConclusionOrlistat treatment improves oxysterol metabolism in overweight and obese adults. Our findings support that orlistat plays a crucial role in the process of endothelial dysfunction and atherosclerosis via oxysterol modulation.
“…Cholesterol uptake and efflux assays were already carried out by different groups, but the described methodology differs in regard to several assay conditions, i.e. differentiation of Caco-2 cells on plates with/without inserts, incubation with cholesterol packaged in micelles or not, composition of the micelles, incubation time and use/no use of acceptors like apoA1 [30][31][32][33][34]. Therefore, we also tested and adjusted these conditions for the cholesterol uptake and efflux assay.…”
Section: Establishing Functional Assays With Caco-2 Cells For Cholestmentioning
Background: The human Caco-2 cell line is a common in vitro model of the intestinal epithelial barrier. As the intestine is a major interface in cholesterol turnover and represents a non-biliary pathway for cholesterol excretion, Caco-2 cells are also a valuable model for studying cholesterol homeostasis, including cholesterol uptake and efflux. Currently available protocols are, however, either sketchy or not consistent among different laboratories. Our aim was therefore to generate a collection of optimized protocols, considering the different approaches of the different laboratories and to highlight possibilities and limitations of measuring cholesterol transport with this cell line. Results: We developed comprehensive and quality-controlled protocols for the cultivation of Caco-2 cells on filter inserts in a single tight monolayer. A cholesterol uptake as well as a cholesterol efflux assay is described in detail, including suitable positive controls. We further show that Caco-2 cells can be efficiently transfected for luciferase reporter gene assays in order to determine nuclear receptor activation, main transcriptional regulators of cholesterol transporters (ABCA1, ABCB1, ABCG5/8, NPC1L1). Detection of protein and mRNA levels of cholesterol transporters in cells grown on filter inserts can pose challenges for which we highlight essential steps and alternative approaches for consideration. A protocol for viability assays with cells differentiated on filter inserts is provided for the first time.
Conclusions:The Caco-2 cell line is widely used in the scientific community as model for the intestinal epithelium, although with highly divergent protocols. The herein provided information and protocols can be a common basis for researchers intending to use Caco-2 cells in the context of cellular cholesterol homeostasis.
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