Category ME L14 High Cholesterol Impairs Water and Gas Transport in Red Blood Cells

Abstract High blood cholesterol is a major health problem, resulting in more than half

a million deaths in the United States alone. It is well established that high

levels of plasma cholesterol are causal to numerous diseases, including

angina, diabetes, and cardiovascular disorders such as the development

of arterial plaque, as a consequence of the long-term effects of high blood

cholesterol. The most abundant molecule in a living cell is water (roughly

70%), which is critical to numerous life processes. Therefore, the precise

and regulated transport and distribution of water in various cellular

compartments is vital to life. Water can be transported slowly through

biological membranes via an osmotic gradient, or rapidly transported

through water channels called aquaporins (AQP). AQP1 was the first

water channel to be discovered, and was first found in the red blood cell

(RBC) membrane. Recent studies demonstrate the involvement of AQP1 in

the transport of CO2 as well as water through the RBC membrane.

Reports further demonstrate the inverse relationship of membrane

cholesterol on O2 transport in RBC. In view of this, the immediate

detrimental role of cholesterol on active water transport, and very likely on

the CO2 and O2 transport in RBC was hypothesized, and tested in the

present study. These studies were performed using isolated rat RBC, and

real-time changes in RBC volume were determined using dynamic light

scattering, following exposure to cholesterol and modulators of the water

channel. Results from the study demonstrate that exposure of RBC to

cholesterol is inhibitory to active water transport via AQP1 in the cell. In

contrast, the removal of cholesterol from RBC membrane using methyl-

β-cyclodextrin potentiates water transport. Recent studies suggest

an increase in PLA2 activity in macrophages of coronary lesions prone to

rupture, and high PLA2 activity in response to osmotic pressure in

cholesterol-containing vesicles. In view of this, the role of PLA2 in RBC

water transport was investigated. Surprisingly, the PLA2 inhibitor ONO-

RS-082 was able to reverse cholesterol’s inhibitory effect on water

transport in RBC, suggesting its potential use in the treatment of

complications resulting from elevated blood cholesterol levels. The present

study demonstrates for the first time the immediate detrimental effects of

cholesterol on RBC function, as well as its amelioration following ONO-RS-

082 treatment. Results from this work are a major advancement in our

understanding and treatment of the immediate detrimental effects of high

cholesterol that likely precede angina, diabetes, and cardiovascular

disorders.

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6. J.H. Steinbach, P.L. Blackshear, R.L. Varco, H. Buchwald. High blood

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