Showing posts with label cholestrol metabolism. Show all posts
Showing posts with label cholestrol metabolism. Show all posts

Tuesday, May 30, 2017

Rose Hip and Heart Disease

Helps Prevent Atherosclerotic Plaques

This study features the use of our ABCA1 Antibody. The ATP-binding cassette transporter (ABC) Protein called ABCA1 has a major impact on cellular and whole body cholesterol metabolism and is likely to play an important role in protecting against cardiovascular disease.

The authors conclude that rose hip is a suitable dietary supplement for preventing atherosclerotic plaques formation by modulating systemic blood pressure and the expression of RCT and inflammatory genes.Michele Cavalera, Ulrika Axling, Catarina Rippe, Karl Swärd, and Cecilia Holm. Dietary rose hip exerts anti-atherosclerotic effects and increases nitric oxide-mediated dilation in ApoE-null mice. The Journal of Nutritional Biochemistry. Available online 21 March 2017. http://dx.doi.org/10.1016/j.jnutbio.2017.02.017...Western blotting. Approximately 50 mg of tissue was homogenized in lysis buffer (50 mM Tris–HCl, pH 7.5, 1 mM EGTA, 1 mM EDTA, 1% NP40, 1 mM Na-orthovanadate, 40 mM NaF, 4 mM Na-pyrophosphate, 0.27 M sucrose, 1 mM DTT, 20 μg/ml leupeptin, 10 μg/ml antipain and 1 μg/ml pepstatin) and subsequently centrifuged at 12,000 g for 20 min at 4 °C. Protein concentration of the supernatant was determined using BCA-assay (Pierce) and 20 μg of proteins were resolved on NuPAGE 4–12% Bis-Tris Gel (Life Technologies – Carlsbad, CA) and electroblotted onto nitrocellulose membranes (Amersham, GE Healthcare – UK). The membranes were washed and incubated with primary and appropriate secondary antibodies. Primary antibodies used were: SR-B1 (ab3 - Abcam), beta-actin (A5441 – Sigma), Abca1 (MO13101 - Neuromics), Abcg1 (orb214917, Biorbyt), Abcg5 (orb5656, Biorbyt), Abcg8 (orb228808, Biorbyt), IL1b (orb101745, Biorbyt), IL6 (orb228222, Biorbyt), NF-kB (ab7970, Abcam) and HSP90 (BD610418, BD bioscience)...

RH supplementation modulates the reverse cholesterol transport pathway. (a) Liver qPCR analysis of cholesterol metabolism showed similar expression of genes involved in the synthesis and conversion of cholesterol into bile acids but increased expression of various RCT genes upon rose hip feeding (n=5–6).
(b) Protein levels of SR-B1, Abcg1 and Abcg5 were increased in the liver of RH-feeding mice (n=6). *P<.05, **P<.01 and ^P=.056 versus CTR.



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Monday, August 11, 2008

Cholesterol Homeostasis and Diabetes

Hormone-sensitive lipase (HSL) is widely expressed in adipose tissue. Interestingly, HSL-null mice have been shown to be resistant to diet-induced obesity. Despite this characteristic, they can also show insulin resistance. This resistance is contributing factor in type 2 diabetes.

Scientists have also shown that pancreatic beta cells, responsible for insulin release, begin to malfunction when their cholesterol levels build up. So what is the link?

The study referenced here suggests that HSL plays a critical role in the hydrolysis of cytosolic cholesteryl esters and that increased levels of hepatic cholesteryl esters, due to lack of action of HSL in the liver, is the main mechanism underlying the imbalance in cholesterol metabolism in HSL-null mice.

Celine Fernandez, Marie Lindholm, Morten Krogh, Stéphanie Lucas, Sara Larsson, Peter Osmark, Karin Berger, Jan Boren, Barbara A Fielding, Keith N. Frayn and Cecilia Holm. Disturbed cholesterol homeostasis in hormone-sensitive lipase null mice. Am J Physiol Endocrinol Metab (July 29, 2008). doi:10.1152/ajpendo.90206.2008.

...Liver samples were homogenized in lysis buffer pH 7.0 containing 25 mM Tris, 150 mM NaCl, 1 mM EDTA, 1% Triton and 1X protease inhibitor cocktail (Complete Mini, Roche). Total protein concentration was measured by BCA assay (Pierce) and 50 μg of protein were subjected to SDS-polyacrylamide gel electrophoresis (8 % polyacrylamide). After transfer to PVDF membranes (Invitrogen), blots were incubated with a primary antibody mouse anti-mouse/rat ABCA1 (Neuromics) according to the instructionsof the manufacturer. As secondary antibody a horseradish peroxidase-conjugated sheep antimouse IgG was used. Western blot analysis was performed using a chemiluminescence system (Luminol) and detection was made using a CCD-camera (LAS 1000, Fuji). Band intensities were quantified using the ImageJ software (http://rsb.info.nih.gov/ij)...