Where is GSH found in the body?
GSH is found in all tissues and body fluids. A healthy balance requires an unequal distribution of GSH and GSSG among these locations,6 similar to the need for sodium and potassium to differ between plasma and cells. In general, the concentrations of GSH within cells are much higher than outside of cells. Nonetheless, the amounts of GSH in the fluids surrounding cells are important because they provide a chemical-defense barrier to protect the cell surfaces.
The total amount of GSH in the body is about 15 grams, of which the cysteine component represents 5 grams. The organs principally responsible for detoxification (ie, the liver and kidneys), have the highest amounts, but the 15 grams are distributed among all major organ systems, including brain, heart, skeletal muscle, intestines, lungs, skin, and the immune system. The liver (6% of the body) has about 4 grams of GSH (25% of the body’s total), which is part of an important homeostatic mechanism. Liver GSH varies as a function of diet, time of day, and body needs.7 The cysteine content of liver GSH is similar to the RDA for sulfur amino acids (methionine plus cysteine), which is 1.4 g for a reference 70 kg individual. Thus, the GSH in the liver is equivalent to a 1-day reserve for the cysteine needed for the body’s protein synthesis.
Homeostatic mechanisms prevent the hepatic GSH content from falling too low.8 During fasting and starvation, GSH and its precursors are derived from muscle and other tissues. Simple calculations show that the entire human body has no more than a 4-day reserve of GSH so that loss of GSH can become critical in catabolic illness or whenever there is a prolonged period of protein/energy insufficiency. Importantly, GSH declines with age9,10 and has a diurnal variation with lowest values in the morning and early afternoon.11 The diurnal variation is linked to cysteine, and cysteine variation increases in individuals over 60 years.11 Thus, older individuals have increased vulnerability in cell injury due to both a decline in total amount of GSH and a decline in its homeostatic control.
Most research has focused on tissue levels of GSH, but the difference between GSH needs and availability may be equally important in the extracellular fluids, which bathe cells. GSH is found in all extracellular biological fluids, including plasma, interstitial fluid, cerebrospinal fluid, alveolar lining fluid, saliva, bile, pancreatic fluid, tears, sweat. and urine.12 The concentration of GSH in body fluids can be up to 1,000-fold lower than found in the tissues, yet all cells appear to release GSH, suggesting a universal requirement for extracellular GSH to protect cell surfaces.12 In addition, specific functions of extracellular GSH are well described. Bile has a high content of GSH to support detoxification of reactive chemicals in the lumen of the small intestines13 and to enhance iron absorption.14 Lipid peroxides are toxic species in the diet that are eliminated by supplemental GSH.15 GSH in the lining fluid of the lungs eliminates airborne oxidants and helps maintain fluidity of the mucus lining the airways. Elimination of bacteria by pulmonary macrophages in vitro is stimulated by added GSH, but this experiment has not been done in humans in vivo.16 GSH also protects human lung cells (in vitro) from influenza virus and protects against influenza in mice.17 One should note that controlled, double-blind studies of these effects have not been done in vivo in humans.
How is GSH Maintained in Tissues and Body Fluids?
GSH is maintained by a continuous cycle of turnover at a rate equivalent to the entire body pool of GSH being made and degraded daily.18 GSH is synthesized from the precursor amino acids (ie, glutamine, glycine, cysteine) in all tissues.19 Cells in certain organs (ie, intestines, lung, kidney) can utilize exogenous GSH by a secondary active transport mechanism.20,21 Supply of GSH from tissue to extracellular fluids occurs through two types of transporters, classified as MRP and OAT transport proteins.22 The molecular nature of the systems that allow transport in the opposite direction (from extracellular spaces into cells) is not known.23 The cycle of GSH release, conversion to precursor amino acids, and resynthesis is termed the “GSH cycle.”3 Although it was earlier proposed that a “γ-glutamyl cycle” functioned in amino acid uptake, this was found to not be an important mechanism. Disulfide forms of GSH include low molecular weight chemicals and protein-bound forms14; under many circumstances, the balance between GSH and these disulfide forms (ie, GSH redox balance) can be more important than the absolute amount of GSH.24
GSH is depleted by elimination of reactive chemicals dependent upon abundant GSH transferases.1 These enzymes increase in response to toxic challenge, and trials have been conducted to determine whether continuous elevation of these enzymes can protect against cancer. In protection against cancer, GSH reacts with cancer-causing chemicals at rates that are faster than the chemical can react with DNA, thereby preventing mutations. To date, however, practical approaches to reduce cancer by increasing GSH transferase have not been established. In addition to cellular activities, GSH transferase is associated with mucus and provides a detoxifying barrier in the small intestines.13 Animal studies showed that provision of GSH to the GSH transferase associated with the mucus provides a defense mechanism to eliminate ingested toxic chemicals, such as oxidation products from polyunsaturated fatty acids, acrylein, acrylamide, and other reactive chemicals, prior to absorption by the body. (Fig 4A) This defense depends upon GSH supply outside of the cells, either from the bile, from food, or from a supplement. The finding that oral and pharyngeal cancer is decreased in association with intake of foods high in GSH25 could reflect the function of this mechanism in protection against cancer-causing chemicals or a better function of the immune system. Studies with human cells in culture further show that added GSH protects cells even in the absence of GSH uptake,26 apparently due to protection of proteins on the surface of cells. Recent studies show that cell surface thiols function as redox sensors, signaling processes such as platelet activation and early events of atherosclerosis.27–29 As indicated above, in vitro experiments have demonstrated that addition of GSH to the media improved killing of bacteria by pulmonary macrophages and decreased production of infectious influenza virus by human small airway epithelial cells.
How Big is the Functional Need for GSH?
In addition to the age-related decline mentioned above, GSH levels are inversely associated with environmental exposures and disease risk. GSH is decreased in the epithelial lining fluid of human lung in individuals who abuse alcohol.30 This example is illustrative of the hidden risks of low GSH in that these individuals have no apparent lung disease and yet are at considerably increased risk of acute lung injury and death from adult respiratory death syndrome.31, 32 Oxidation of GSH occurs in association with increased carotid intima media thickness, an indicator of cardiovascular disease risk.33 GSH redox balance (ie, the GSH/ GSSG ratio) favors oxidation in cigarette smokers34 and type 2 diabetics.35 Direct evidence that the decrease and oxidation of GSH occurs due to toxic chemical exposures is available from studies in individuals following chemotherapy.36 The extensive evidence that GSH status is decreased in association with disease and recognized risk factors for disease implies that maintenance of this protective system could reduce risk of disease development.
Because of the known functions and increased disease risk with a decline of GSH, systematic efforts are needed to quantify the difference between the available GSH and the amount needed. One approach is to consider how much GSH is present in a natural diet. GSH content has been measured in more than 100 common foods37 and provides the basis to estimate dietary intake. The best diets contain about 150 milligrams of GSH per day; the worst diets contain as little as 3 milligrams per day.37 GSH is present in essentially all raw and freshly prepared foods; the best sources are fresh fruits and vegetables, nuts, and whole-cut meats, including poultry and fish. GSH can also be increased by supplements, such as the increase in hepatic GSH following ingestion of silymarin, found in milk thistle. GSH is lost during most food processing procedures, with the exception of fresh-frozen foods. Processed, cured, and canned meat products have essentially no GSH. Similarly, canned or dried fruits and canned vegetables are not good sources. Cereal and grain products are largely deficient, and almost all dairy products, beverages, sweeteners, and condiments lack GSH. Thus, a simple conclusion is that modern processed foods are deficient in GSH compared to natural, freshly prepared foods.37 In quantitative terms, up to 150 mg of daily intake of GSH can be lost due to food processing.
Many foods also contain reactive chemicals that remove GSH through the GSH transferase reaction associated with the lining of the small intestines. Measurement of a broad range of foods show that milk, prunes, tea, blueberries, and bottled apple juice have high contents of GSH-reactive chemicals.38 Recently, there has been interest in the potent neurotoxicant acrylamide, because this has been found to be relatively high in french fries.39 The daily intake of GSH-reactive equivalents can range from almost zero to values exceeding the maximum naturally available 150 milligrams GSH.38 Thus, the sum of the amount of GSH needed to eliminate reactive chemicals and the amount of GSH lost by food processing can be greater than 300 mg of GSH per day.
Where is GSH found in the body?
GSH is found in all tissues and body fluids. A healthy balance requires an unequal distribution of GSH and GSSG among these locations,6 similar to the need for sodium and potassium to differ between plasma and cells. In general, the concentrations of GSH within cells are much higher than outside of cells. Nonetheless, the amounts of GSH in the fluids surrounding cells are important because they provide a chemical-defense barrier to protect the cell surfaces.
The total amount of GSH in the body is about 15 grams, of which the cysteine component represents 5 grams. The organs principally responsible for detoxification (ie, the liver and kidneys), have the highest amounts, but the 15 grams are distributed among all major organ systems, including brain, heart, skeletal muscle, intestines, lungs, skin, and the immune system. The liver (6% of the body) has about 4 grams of GSH (25% of the body’s total), which is part of an important homeostatic mechanism. Liver GSH varies as a function of diet, time of day, and body needs.7 The cysteine content of liver GSH is similar to the RDA for sulfur amino acids (methionine plus cysteine), which is 1.4 g for a reference 70 kg individual. Thus, the GSH in the liver is equivalent to a 1-day reserve for the cysteine needed for the body’s protein synthesis.
Homeostatic mechanisms prevent the hepatic GSH content from falling too low.8 During fasting and starvation, GSH and its precursors are derived from muscle and other tissues. Simple calculations show that the entire human body has no more than a 4-day reserve of GSH so that loss of GSH can become critical in catabolic illness or whenever there is a prolonged period of protein/energy insufficiency. Importantly, GSH declines with age9,10 and has a diurnal variation with lowest values in the morning and early afternoon.11 The diurnal variation is linked to cysteine, and cysteine variation increases in individuals over 60 years.11 Thus, older individuals have increased vulnerability in cell injury due to both a decline in total amount of GSH and a decline in its homeostatic control.
Most research has focused on tissue levels of GSH, but the difference between GSH needs and availability may be equally important in the extracellular fluids, which bathe cells. GSH is found in all extracellular biological fluids, including plasma, interstitial fluid, cerebrospinal fluid, alveolar lining fluid, saliva, bile, pancreatic fluid, tears, sweat. and urine.12 The concentration of GSH in body fluids can be up to 1,000-fold lower than found in the tissues, yet all cells appear to release GSH, suggesting a universal requirement for extracellular GSH to protect cell surfaces.12 In addition, specific functions of extracellular GSH are well described. Bile has a high content of GSH to support detoxification of reactive chemicals in the lumen of the small intestines13 and to enhance iron absorption.14 Lipid peroxides are toxic species in the diet that are eliminated by supplemental GSH.15 GSH in the lining fluid of the lungs eliminates airborne oxidants and helps maintain fluidity of the mucus lining the airways. Elimination of bacteria by pulmonary macrophages in vitro is stimulated by added GSH, but this experiment has not been done in humans in vivo.16 GSH also protects human lung cells (in vitro) from influenza virus and protects against influenza in mice.17 One should note that controlled, double-blind studies of these effects have not been done in vivo in humans.
How is GSH Maintained in Tissues and Body Fluids?
GSH is maintained by a continuous cycle of turnover at a rate equivalent to the entire body pool of GSH being made and degraded daily.18 GSH is synthesized from the precursor amino acids (ie, glutamine, glycine, cysteine) in all tissues.19 Cells in certain organs (ie, intestines, lung, kidney) can utilize exogenous GSH by a secondary active transport mechanism.20,21 Supply of GSH from tissue to extracellular fluids occurs through two types of transporters, classified as MRP and OAT transport proteins.22 The molecular nature of the systems that allow transport in the opposite direction (from extracellular spaces into cells) is not known.23 The cycle of GSH release, conversion to precursor amino acids, and resynthesis is termed the “GSH cycle.”3 Although it was earlier proposed that a “γ-glutamyl cycle” functioned in amino acid uptake, this was found to not be an important mechanism. Disulfide forms of GSH include low molecular weight chemicals and protein-bound forms14; under many circumstances, the balance between GSH and these disulfide forms (ie, GSH redox balance) can be more important than the absolute amount of GSH.24
GSH is depleted by elimination of reactive chemicals dependent upon abundant GSH transferases.1 These enzymes increase in response to toxic challenge, and trials have been conducted to determine whether continuous elevation of these enzymes can protect against cancer. In protection against cancer, GSH reacts with cancer-causing chemicals at rates that are faster than the chemical can react with DNA, thereby preventing mutations. To date, however, practical approaches to reduce cancer by increasing GSH transferase have not been established. In addition to cellular activities, GSH transferase is associated with mucus and provides a detoxifying barrier in the small intestines.13 Animal studies showed that provision of GSH to the GSH transferase associated with the mucus provides a defense mechanism to eliminate ingested toxic chemicals, such as oxidation products from polyunsaturated fatty acids, acrylein, acrylamide, and other reactive chemicals, prior to absorption by the body. (Fig 4A) This defense depends upon GSH supply outside of the cells, either from the bile, from food, or from a supplement. The finding that oral and pharyngeal cancer is decreased in association with intake of foods high in GSH25 could reflect the function of this mechanism in protection against cancer-causing chemicals or a better function of the immune system. Studies with human cells in culture further show that added GSH protects cells even in the absence of GSH uptake,26 apparently due to protection of proteins on the surface of cells. Recent studies show that cell surface thiols function as redox sensors, signaling processes such as platelet activation and early events of atherosclerosis.27–29 As indicated above, in vitro experiments have demonstrated that addition of GSH to the media improved killing of bacteria by pulmonary macrophages and decreased production of infectious influenza virus by human small airway epithelial cells.
How Big is the Functional Need for GSH?
In addition to the age-related decline mentioned above, GSH levels are inversely associated with environmental exposures and disease risk. GSH is decreased in the epithelial lining fluid of human lung in individuals who abuse alcohol.30 This example is illustrative of the hidden risks of low GSH in that these individuals have no apparent lung disease and yet are at considerably increased risk of acute lung injury and death from adult respiratory death syndrome.31, 32 Oxidation of GSH occurs in association with increased carotid intima media thickness, an indicator of cardiovascular disease risk.33 GSH redox balance (ie, the GSH/ GSSG ratio) favors oxidation in cigarette smokers34 and type 2 diabetics.35 Direct evidence that the decrease and oxidation of GSH occurs due to toxic chemical exposures is available from studies in individuals following chemotherapy.36 The extensive evidence that GSH status is decreased in association with disease and recognized risk factors for disease implies that maintenance of this protective system could reduce risk of disease development.
Because of the known functions and increased disease risk with a decline of GSH, systematic efforts are needed to quantify the difference between the available GSH and the amount needed. One approach is to consider how much GSH is present in a natural diet. GSH content has been measured in more than 100 common foods37 and provides the basis to estimate dietary intake. The best diets contain about 150 milligrams of GSH per day; the worst diets contain as little as 3 milligrams per day.37 GSH is present in essentially all raw and freshly prepared foods; the best sources are fresh fruits and vegetables, nuts, and whole-cut meats, including poultry and fish. GSH can also be increased by supplements, such as the increase in hepatic GSH following ingestion of silymarin, found in milk thistle. GSH is lost during most food processing procedures, with the exception of fresh-frozen foods. Processed, cured, and canned meat products have essentially no GSH. Similarly, canned or dried fruits and canned vegetables are not good sources. Cereal and grain products are largely deficient, and almost all dairy products, beverages, sweeteners, and condiments lack GSH. Thus, a simple conclusion is that modern processed foods are deficient in GSH compared to natural, freshly prepared foods.37 In quantitative terms, up to 150 mg of daily intake of GSH can be lost due to food processing.
Many foods also contain reactive chemicals that remove GSH through the GSH transferase reaction associated with the lining of the small intestines. Measurement of a broad range of foods show that milk, prunes, tea, blueberries, and bottled apple juice have high contents of GSH-reactive chemicals.38 Recently, there has been interest in the potent neurotoxicant acrylamide, because this has been found to be relatively high in french fries.39 The daily intake of GSH-reactive equivalents can range from almost zero to values exceeding the maximum naturally available 150 milligrams GSH.38 Thus, the sum of the amount of GSH needed to eliminate reactive chemicals and the amount of GSH lost by food processing can be greater than 300 mg of GSH per day.
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