BASIC NUTRITIONAL INVESTIGATION

Nutrition Vol. 14, No. 3, 1998

Dose- and Time-Dependent Hypocholesterolemic Effect of Oyster Mushroom (Pleurotus ostreatus) in Rats
R OZDI N, PHD,* AND S TEFAN GALBAVY , MD PAVEL BOBEK, PHD,* LUBOMI From the *Research Institute of Nutrition, and the Faculty of Medicine, University of J.A. Komensky , Bratislava, Slovak Republic Date accepted: 30 July 1997
ABSTRACT

The effect of the dose of oyster mushroom in the diet (1.0, 2.5, and 5.0%) and of the period of application (8, 16, 28, and 52 wk) on cholesterol accumulation in blood and body organs was studied in weanling male Wistar rats fed a diet containing 0.3% cholesterol. Reduction of cholesterol in serum and body organs was found to be dependent on the amount of dietary oyster mushroom administered. A negative correlation between the mushroom dose and cholesterol level was found after 8 and 28 wk of feeding (r 0.9821 and 0.9803, respectively; P 0.02 for both cases). The dose of 1% oyster mushroom did not affect cholesterol levels in serum or body organs. A signicant reduction of cholesterol levels was observed in serum (31 46%) and liver (2530%) at a dose of 5% of oyster mushroom for all periods. Reduced cholesterol content in very low-density lipoproteins (VLDL) was also observed at this level. The highest dose of oyster mushroom induced a decrease in conjugated diene levels in erythrocytes and an increase in the levels of reduced glutathione in the liver and stimulated the activities of catalase and glutathione peroxidase in the liver in the nal period of the experiment. Nutrition 1998;14:282286. Elsevier Science Inc. 1998 Key words: rat, oyster mushroom, cholesterol, lipoperoxidation, dose-dependency, time-dependency

INTRODUCTION

It is generally accepted that lowering of high serum cholesterol levels plays a signicant role in the prevention of atherosclerosis.1 The search for natural substances with hypocholesterolemic effects is, therefore, desirable particularly in countries with a persistent incidence of hypercholesterolemia and cardiovascular disease. Higher fungi are an ideal dietary substance for the prevention and treatment of hypercholesterolemia due to the high content of dietary ber, protein and microelements, and the presence of plant sterols, as well as the low energy content. Oyster mushroom (Pleurotus ostreatus) is a wood-rotting fungus produced on lignocellulose substrates in a number of countries on a large scale for the food industry. We have found in a series of experiments that long-term dietary supplementation with 5% dried oyster mushroom fruiting bodies can effectively suppress dietary-induced hypercholesterolemia in rats.2 A prospective clinical trial using dried oyster mushroom as a natural hypolipidemic agent prompted this study of the hypolipidemic effect of oyster mushroom in rats with dietary hypercholesterolemia as a function of the administration

period and dose. In concert with the proven role of oxidative stress in the etiology of atherogenesis,3 we also examined the effect of long-term administration of oyster mushroom on the oxidativeantioxidative status in the rat.
MATERIAL AND METHODS

Weanling male Wistar rats (n 112) with initial body weights of approximately 60 g were used. Control animals were fed ad libitum a semisynthetic diet4 of the following composition (g/100 g): starch 60, casein 18, pork fat 10, cellulose 6, mineral and vitamin mixtures 4 and 1, respectively, Fel tauri (commercial dried ox bile) 0.55, cholesterol 0.3, and choline chloride 0.15. The animals in experimental mushroom groups 13 were fed the diet with aliquots of cellulose substituted with 1.0, 2.5, and 5.0% powdered, dried fruiting bodies of oyster mushroom. Animals were killed by decapitation (in light ether narcosis after an 18 h fast) in intervals of 8, 16, 28, and 52 wk. Cholesterol in serum and lipoproteins (isolated by sequential otation5) and serum triacylglycerols were estimated by enzy-

Correspondence to: Pavel Bobek, PhD, Research Institute of Nutrition, Limbova 14, Bratislava 833 37, Slovak Republic.

Nutrition 14:282286, 1998 Elsevier Science Inc. 1998 Printed in the USA. All rights reserved.

0899-9007/98/$19.00 PII S0899-9007(97)00471-1

HYPOCHOLESTEROLEMIC EFFECT OF OYSTER MUSHROOM TABLE I.

283

THE DOSE- AND TIME-DEPENDENT EFFECT OF OYSTER MUSHROOM ON CHOLESTEROL AND TRIACYLGLYCEROL CONTENT IN RAT SERUM AND ORGANS Diet Interval/Parameter 8 wk Body weight (g) Cholesterol Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) Aorta (mmol kg1) Triacylglycerols Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) 16 wk Body weight (g) Cholesterol Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) Aorta (mmol kg1) Triacylglycerols Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) 28 wk Body weight (g) Cholesterol Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) Aorta (mmol kg1) Triacylglycerols Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) 52 wk Body weight (g) Cholesterol Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) Aorta (mmol kg1) Triacylglycerols Serum (mmol L1) Liver (mmol kg1) Heart (mmol kg1) Control Mushroom 1 Mushroom 2 Mushroom 3

337 13 5.60 0.60 326 19 7.60 0.62 4.04 0.15 0.43 0.04 37.33 5.99 3.94 0.39 401 12 5.39 0.41 540 27E 7.32 0.36 5.10 0.20D 0.56 0.05 28.04 4.15 2.54 0.50 446 16 6.81 0.43 537 20E 6.83 0.35 6.03 0.23E 0.34 0.06 70.19 5.48D 3.14 0.62 562 14 7.56 1.07 474 35C 5.53 0.32C 6.12 0.39E 0.60 0.06A 54.31 4.16A 3.54 0.56

300 12 4.95 0.24 327 19 6.02 0.30 3.96 0.14 0.54 0.04 42.93 10.92 3.89 0.98 414 14 4.60 0.59 527 26E 6.29 0.27a 4.91 0.24C 0.61 0.06 35.11 7.13 3.55 0.76 463 13 5.34 0.54 489 14E 6.14 0.29 6.10 0.19E 0.39 0.08 55.58 3.58a 2.86 0.51 587 25 5.78 0.57 403 17C 5.84 0.14 5.98 0.28E 0.74 0.12 34.47 3.05C 3.24 0.85

314 12 3.84 0.27C 276 15 6.65 0.70 3.58 0.10a 0.37 0.02 32.63 5.68 3.47 0.45 413 20 4.35 0.53 431 29b,E 6.03 0.12c 5.18 0.22E 0.67 0.05E 26.03 6.05 2.83 0.37 457 13 5.05 0.62a 479 26E 5.96 0.18a 5.14 0.16c,E 0.39 0.03 54.36 3.65a,C 2.56 0.42 545 18 4.40 0.37b 311 32c 5.40 0.88 6.03 0.39E 0.61 0.03E 30.50 0.82e 2.77 0.30

315 13 3.04 0.25d 232 12d 5.11 0.11c 3.62 0.09d 0.44 0.03 36.96 9.95 4.22 0.66 398 10 3.24 0.24e 398 27c,E 5.86 0.15c 5.13 0.16E 0.47 0.03 28.58 4.88 2.21 0.30 448 16 4.73 0.66a,A 376 18e,E 5.42 0.25c 5.42 0.16a,E 0.46 0.05 51.80 3.74b 3.02 0.56 556 17 4.38 0.61a 355 28b,D 4.69 0.78 5.45 0.24E 0.70 0.04E 32.22 3.41d 2.67 0.24A

Values are means SEM for 7 animals in each group. a e Statistical signicance of mushroom groups versus control group; AE statistical signicance of values in 16, 28, and 52 wk versus values in 8 wk: a,A P 0.05; b,B P 0.02; c,C P 0.01; d,D P 0.002; e,E P 0.001. Mushroom 1, 1.0%; Mushroom 2, 2.5%; Mushroom 3, 5.0% oyster mushroom.

matic kit (Oxochrom Chol 250 E, Czech Republic). Cholesterol content in individual tissues (liver, heart, and aorta) was quantied by Bio-La-Test kit (Lachema, Czech Republic) after chloroformmethanol (2:1) extraction.6 The content of conjugated dienes in plasma, erythrocytes and in liver was assayed according to

Recknagel et al.7 The following antioxidative parameters were estimated in erythrocytes and liver: total antioxidant status (Randox Lab, Ltd., Crumlin, Northern Ireland, UK); reduced glutathione level (GSH)8; activities of superoxide dismutase (SOD) (Randox Lab Ltd.), catalase (CAT)9, and glutathione peroxidase (GSH-

284

HYPOCHOLESTEROLEMIC EFFECT OF OYSTER MUSHROOM TABLE II.

CHOLESTEROL DISTRIBUTION IN LIPOPROTEINS OF RATS Diet Interval/Lipoprotein 8 wk VLDL (mmol (%)* LDL (mmol (%) HDL (mmol (%) 16 wk VLDL (mmol (%) LDL (mmol (%) HDL (mmol (%) 28 wk VLDL (mmol (%) LDL (mmol (%) HDL (mmol (%) 52 wk VLDL (mmol (%) LDL (mmol (%) HDL (mmol (%) Control Mushroom 1 Mushroom 2 Mushroom 3

L1) L1) L1)

3.14 0.43 55.0 3.2 1.69 0.33 29.5 3.2 0.86 0.09 15.6 5.0 2.82 0.29 55.7 1.3 1.44 0.09 28.3 0.7 0.79 0.07 16.0 1.0 3.78 0.34 55.3 2.8 1.76 0.17 28.8 2.2 1.23 0.23 18.2 3.1 3.63 0.68 47.8 4.5 1.90 0.26 26.7 1.0 1.68 0.1A 25.6 3.9

2.75 0.17 59.0 2.0 1.11 0.17 23.2 2.4 0.83 0.04 17.9 0.6 2.40 0.35 49.3 2.0C 1.52 0.20 30.4 2.6 0.91 0.02 20.2 3.1 2.42 0.15c 46.7 3.0C 1.48 0.28 29.8 4.5 1.38 0.22 26.2 3.7A 2.12 0.65 43.2 3.5C 2.65 0.86 43.2 3.9d,E 1.37 0.06 22.3 5.4

1.88 0.22a 51.5 2.2 0.85 0.12a 23.3 1.5 0.88 0.10 25.3 3.6 2.22 0.34 48.5 3.4 1.23 0.19 27.3 2.3 1.05 0.05 24.2 2.2c 2.50 0.31a 50.6 1.3 1.63 0.44 29.7 3.3 0.96 0.16 19.7 3.0 2.32 0.36 49.0 3.1 1.26 0.16 27.0 1.1 1.10 0.14 24.0 2.5

1.06 0.12e 34.5 1.2 0.75 0.18a 24.3 2.1 1.27 0.12b 41.2 3.2d 1.73 0.14c,C 48.3 3.2D 0.62 0.10e 17.0 1.9e,A 1.23 0.14b 34.7 4.2e 2.0 0.43c 43.0 0.8d,E 1.76 0.38A 37.8 3.99C 0.89 0.13 19.1 2.8D 1.58 0.18b,A 37.4 3.8 1.29 0.14A 30.5 4.2 1.36 0.16 32.2 6.9

L1) L1) L1)

L1) L1) L1)

L1) L1) L1)

a e, AE

Values are means SEM for 7 animals in each group. Statistical signicance as in Table I. * Percent of total serum cholesterol. Mushroom 1, 1.0%; Mushroom 2, 2.5%; Mushroom 3, 5% oyster mushroom.

Px).10 Protein content in liver homogenates was determined according to Lowry et al.11 Samples from heart and liver were taken for histological examination. The results were statistically evaluated by Students t test.
RESULTS

The body weight of experimental animals was not affected by the presence of oyster mushroom in the diet for all doses and time periods tested. A time-dependent increase of cholesterol levels in serum, liver, and aorta was observed in cholesterol diet-fed animals. Analogous changes in triacylglycerol levels were less pronounced and showed no clear trend. The diet supplemented with 1% oyster mushroom did not signicantly affect the levels of serum cholesterol or triacylglycerols in any of the investigated intervals. A signicant reduction of cholesterol levels could be conrmed only in heart in week 16, and reduction of triacylglycerol levels in liver in weeks 28 and 52. The diet with 2.5% oyster mushroom signicantly reduced cholesterol levels in serum after 8, 28, and 52 wk, in liver after 16 and 52 wk, in heart after 16 and 28 wk, and in aorta after 8 and 28 wk. Diets supplemented with 5% oyster mushroom signicantly reduced cholesterol levels in serum (31 46%) and in liver (2530%) in all examined periods.

The cholesterol content in heart was also signicantly reduced for all intervals, with the exception of the last one, while in aorta it decreased after 8 and 28 wk. Triacylglycerol levels in serum and body organs were not affected at any dose or time period tested (with the exception of the reduced content in liver for all oyster mushroom doses after 28 and 52 wk) (Table I). The cholesterol diet modied the cholesterol distribution in lipoproteins in all tested time periods. This was observed as a shift from high-density lipoproteins (HDL) (which are dominant under physiologic conditions containing 60 70% of cholesterol) to very low-density lipoproteins (VLDL) (which are dominant under feeding a cholesterol-rich diet and carrying 40 60% of total cholesterol). The lowest oyster mushroom dose did not signicantly alter cholesterol content in any lipoprotein class. The dose of 2.5% of oyster mushroom in the diet caused a signicant reduction of cholesterol in VLDL after 8 and 28 wk, in low-density lipoproteins (LDL) after 8 wk only, and HDL cholesterol level was increased after 16 wk only. The diet supplemented with 5% oyster mushroom reduced VLDL cholesterol levels (40 60%) in all tested periods. LDL cholesterol was reduced (nearly 60%) after 8 and 16 wk of treatment with a simultaneous increase of HDL cholesterol concentration (48 and 56%, respectively) in the same

HYPOCHOLESTEROLEMIC EFFECT OF OYSTER MUSHROOM TABLE III.

CONCENTRATION OF CONJUGATED DIENES, TOTAL ANTIOXIDANT CAPACITY, REDUCED GLUTATHIONE LEVELS AND OF ANTIOXIDANT ENZYME ACTIVITY IN PLASMA ERYTHROCYTES AND LIVER OF RATS AFTER 52 WEEKS Diet Parameter Conjugated dienes* Erythrocytes (d mL1) Plasma (d mL1) Liver (d g1) TAS* Plasma (mmol L1) Liver (mmol g1) SOD** Erythrocytes (U mL1) Liver (U mg1) CAT** Erythrocytes (U mL1) Liver (U mg1) GSH-Px** Erythrocytes (U mL1) Liver (U mg1) GSH** Erythrocytes (mol mL1) Liver (mol mg1) Control Mushroom 1 Mushroom 2 Mushroom 3

285

15.64 1.16 0.62 0.09 26.23 1.75 2.69 0.11 0.099 0.013 229 11 20.03 3.54 7410 867 19.34 0.36 13.13 1.32 0.099 0.006 0.26 0.011 2.71 0.13

13.75 2.28 0.66 0.11 26.76 1.16 2.54 0.07 0.047 0.013b 234 10 11.31 1.69a 7298 859 21.49 0.47C 13.61 1.22 0.098 0.011 0.25 0.010 2.66 0.24

7.71 1.43d 0.69 0.13 23.43 1.60 3.17 0.22 0.081 0.007 239 7 25.69 5.11 4910 607a 19.85 1.21 17.62 1.10 0.135 0.020 0.25 0.006 2.93 0.12

10.80 1.33 0.73 0.04 32.78 3.49 3.11 0.18 0.076 0.01 228 19 30.37 5.93 5676 353 23.37 1.64 22.26 0.82 0.222 0.01 0.28 0.02 3.41 0.28

Values are means SEM for 7 animals in each group. * Values are expressed per milliliter of blood and plasma, or per gram of tissue. ** Values are expressed per milliliter of blood or per milligram of tissue. a e Statistical signicance of mushroom groups versus control group as in Table I. Mushroom 1, 1.0%; Mushroom 2, 2.5%; Mushroom 3, 5.0% oyster mushroom. TAS, total antioxidant capacity; SOD, superoxide dimutase; CAT, catalase; GSH-Px, glutathione peroxidase; GSH, glutathione.

intervals. The amount of cholesterol carried by VLDL and LDL (percentage of total cholesterol) was signicantly reduced only in the animals fed a diet containing 5% oyster mushroom (for VLDL in week 8 and 28, for LDL only in week 16). The contribution of HDL to cholesterol distribution was signicantly higher only for 5% oyster mushroom in the diet and just for weeks 8 and 16 (Table II). Upon analysis, conjugated diene content was found significantly reduced in erythrocytes of animals fed the diet with 2.5 and 5.0% oyster mushroom in the last period of the experiment. However, no changes in conjugated dienes in plasma or liver were observed. The diet with 1% oyster mushroom reduced the activity of SOD and the total antioxidative capacity in liver; higher doses of oyster mushroom returned these parameters to values found in the control animals. The lowest dose of oyster mushroom stimulated CAT activity in liver. However, the activity of this enzyme in erythrocytes was reduced by the 2.5% oyster mushroom diet. A signicant increase in the activity of CAT in liver, GSH-Px in erythrocytes and liver, and the level of reduced glutathione in liver was observed at the dose of 5% oyster mushroom (Table III). The histologic examination did not reveal any effect of dietary oyster mushroom on the histology of myocardium, irrespective of the mushroom dose. Steatosis was the dominant nding in the liver. However, it was less pronounced in groups with lower cholesterol and triacylglycerol levels in liver, i.e., in groups with higher oyster mushroom content in the diet.

DISCUSSION

The results of the study clearly proved that the capacity of oyster mushroom to reduce cholesterol levels in serum and in tissues is dose-dependent. We have found a negative correlation between serum cholesterol level and the dose of oyster mushroom after 8 and 28 wks of treatment (r 0.9821 and 0.9803; P 0.02 for both cases). In spite of progressive hypercholesterolemia, the reduction of serum cholesterol was less signicant after 28 and 52 wk on inclusion of 5% oyster mushroom in the diet. A less signicant reduction of cholesterol content was also observed in the organs (especially in aorta and heart) in the nal phases of the experiment. This less efcient reduction of cholesterolemia by oyster mushroom is related to a retarded decrease in VLDL cholesterol and to the absence of a decrease in LDL-cholesterol in this phase of the experiment. Our data do not provide any rational explanation for this phenomenon. The fact that rat liver LDL receptor function is not affected by a cholesterol diet12 can certainly contribute to an effective control of cholesterolemia in rat. However, this effect results in a massive accumulation of cholesterol in liver. It can be hypothesized that a limiting saturation of the receptor system is reached near the end of the experiment, which results in the failure of the mechanisms controlling LDL cholesterol and cholesterolemia. In addition, the liver plays a key role in the regulation of cholesterol metabolism and a serious steatosis can result in the loss of its functions. Liver steatosis is accompanied by a massive accumulation of cholesterol leading to a nearly 40-fold enhancement of cholesterol content in the liver com-

286 pared to the control animals on a cholesterol-free diet while triacylglycerol accumulation is less than two-fold. The aforementioned arguments may indicate that cholesterol removal from the liver is progressively restricted near the end of the experiment resulting in at least a 25- to 28-fold enhancement of its liver content. This is in contrast to the return of triacylglycerol content to physiological levels. Our earlier results from a shorter 8-wk experiment showed that oyster mushroom in the diet was able to reduce the formation of cholesterol-enriched VLDL13 (which are quantitatively the most signicant lipoprotein class in the control of serum cholesterol levels) through the inhibition of cholesterol absorption14 and biosynthesis,15 VLDL production,13 and acceleration of their fractional turnover.2 Because oyster mushroom evidently affects several steps in the regulation of cholesterol metabolism, it would be of interest to investigate whether preventive administration of oyster mushroom could stimulate the ability of the rat to cope with a high cholesterol diet. Powdered oyster mushroom contains: polysaccharides (6570%), proteins (20 25%), lipids (2.2%), ash (4.8%), and water (up to 5%). Oyster mushroom contains a number of substances with a potential effect on the regulation of cholesterol metabolism. One of these substances is mevinolin16 (lovastatin, the rst from a series of highly active hypocholesterolemic drugs-statins), which inhibits HMG-CoA reductase, the key enzyme of cholesterol biosynthesis in liver, and increases the activity of liver LDL receptors.17 The brous matter in oyster mushroom, particularly its water-soluble components (beta-1,3-Dglucan with low degree of polymerization) and pectin (1520 and 6% of dry matter, respectively) are able to bind bile acids, therefore, reducing the formation of micelles18 and cholesterol absorp-

HYPOCHOLESTEROLEMIC EFFECT OF OYSTER MUSHROOM tion. Higher excretion of bile acids17 induces a decrease of their enterohepatic circulation and, by a feedback mechanism, the stimulation of 7 alpha-hydroxylase,19 the rate-limiting enzyme in the catabolism of cholesterol to bile acids. Oxidative stress, as the disturbed balance between oxidative and antioxidative processes in favor of the former, plays a signicant role in the etiology of atherosclerosis.3 In addition to proatherogenic redistribution of the lipoprotein prole (enrichment of VLDL and LDL at the expense of HDL, which are the dominant cholesterol carriers under physiological conditions), a cholesterol-rich diet resulted in increased lipid peroxidation with simultaneous reduced activity of glutathionedependent antioxidant defense enzymes GSH-Px and glutathione transferase.20 Particularly important with respect to atherogenesis is the nding that a cholesterol diet increases the sensitivity of LDL to peroxidation21 and induces higher production of toxic superoxide radicals in the arterial endothelium in rabbits.22 It can be supposed that reduced content of conjugated dienes (primary products of lipid peroxidation) in erythrocytes and increased activities of antioxidant enzymes CAT and GSH-Px (together with increased content of GSH as cofactor) in liver are favorable antiatherogenic effects of a diet supplemented with 5% oyster mushroom. We do not have data explaining the mechanisms of antioxidant effects of oyster mushroom. It has been found that lowpolymerized water-soluble beta-glucans are able to quench free radicals.23 It is interesting in this respect that hypocholesterolemic drugs (that actually work analogously to oyster mushroom either by sequestration of bile acids or by inhibition of HMG-CoA reductase) reduced the level of lipid peroxides in serum and liver, and increased the resistance of LDL and liver lipids to peroxidation.24 The results obtained so far in our laboratory show a potential for the use of oyster mushroom as a component in the prevention of dietary-induced hypercholesterolemia.

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1. Stamler J, Welthwoth D, Neaton JD. Is there a relationship between serum cholesterol and risk of premature death from coronary heart disease and grade? J Am Med Assoc 1986;253:2823 , Ozd . The mushroom Pleurotus ostreatus 2. Bobek P, Kuniak L n L reduces secretion and accelerates the fractional turnover rate of verylow-density lipoproteins in rat. Ann Nutr Metab 1993;37:142 3. Steinberg D, Parthasarathi S, Carew TE, Khoo JC, Witzum JL. Mechanism of disease. Beyond cholesterol (Modications of low-density lipoprotein that induces atherogenicity). N Engl J Med 1989;320:915 4. Yamashita S, Yamashita K, Yasuda H. High-bre diet in the control of diabetes in rats. Endocrinol Jpn 1980;27:169 5. Havel RJ, Eder HA, Bragdon JH. The distribution of ultracentrifugally separated lipoproteins in human serum. J Clin Invest 1955;34:1345 6. Folch J, Lees M, Sloane-Stanley GH. A simple method for the isolation and purication of total lipids from animal tissue. J Biol Chem 1957;226:497 7. Recknagel R, Glende EA. Spectrophotometric detection of lipid conjugated dienes. In: Colowick SR, Kaplan NO, eds. Methods in enzymology. San Diego: Academic Press, 1984:331 8. Akerboom TPM, Sies H. Assay of glutatione disulde and glutatione mixed disuldes in biological samples. In: Jacoby WB, ed. Methods in enzymology, New York: Academic Press, 1981:373 9. Cavarocchi NC, England MD, OBrien JF. Superoxide generation during cardiopulmonary bypassis there a role for vitamin E. J Surg Res 1986;40:519 10. Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterisation of erythrocyte glutatione peroxidase. J Lab Clin Med 1978;70:158 11. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurment with the Folin phenol reagent. J Biol Chem 1951;90:265 12. Havel RJ. Cholesterol transport in lipoproteins. Atherosclerosis Review 1991;23:1 . Oyster mushroom (Pleurotus ostreatus) reduces 13. Bobek P, Ozd n L the production and secretion of very- low-density lipoproteins in hypercholesterolemic rats. Z Ernahrungswiss 1996;35:249 , Kuniak L. Mechanism of hypocholesterolemic 14. Bobek P, Ozd n L effect of oyster mushroom (Pleurotus ostreatus) in rats: reduction of cholesterol absorption and increase of plasma cholesterol removal. Z Ernahrungswiss 1994;33:44 . Oyster mushroom (Pleurotus 15. Bobek P, Hromadova M, Ozd n L ostreatus) reduces the activity of 3-hydroxy-3-methylglutaryl CoA reductase in rat liver microsomes. Experientia 1995;51:589 16. Gunde-Cimerman N, Friedrich J, Cimerman A, Benic ki N. Screening fungi for the production of an inhibitor of HMG CoA reductase: production of mevinolin by the fungi of the genus Pleurotus. FEMS Microbiol Lett 1993;111:203 17. Fidge NH. Fighting high cholesterol levels-lipid lowering drugs. Med J Austral 1993;159:815 18. Vahouny GV, Tombes R, Cassidy MM, Kritchevsky D, Gallo L. Dietary bers. V. Binding of bile salts, phospholipids and cholesterol from mixed micells by bile acid sequestrans and dietary bers. Lipid 1980;15:1012 . Oyster mushroom 19. Bobek P, Ondreic ka R, Klvanova J, Ozd n L (Pleurotus ostreatus) decreases serum and liver cholesterol and increases cholesterol 7 alpha-hydroxylase activity and fecal excretion of neutral sterols and bile acids in hyper cholesterolemic rats. Nutr Res 1994;14:1683 20. Uysal M, Kutalp G, Seckin S. The effect of cholesterol feeding on lipid peroxide, glutathione, glutathione peroxidase and glutathione transferase in the liver of rats. Int J Vit Nutr Res 1988;58:339 21. Nenseter MS, Gudmundsen O, Malterud KE, Berg T, Drevon CA. Effect of cholesterol feeding on the susceptibility of lipoproteins to oxidative modication. Biochim Biophys Acta 1994;1213:207 22. OHara Y, Peterson E, Harrison DG. Hypercholesterolemia increases endothelial superoxide anion production. J Clin Invest 1993;91:2546 23. Klurfeld DM. Dietary ber mediated mechanisms in carcinogenesis. Cancer Res 1992;52:255 24. Hoffman R, Brook GJ, Aviram M. Hypolipemic drugs reduce lipoprotein susceptibility to undergo lipid peroxidation: in vitro and in vivo studies. Atherosclerosis 1992;93:105