Comparative effects of curcumin and photo-irradiated curcumin on alcohol-and polyunsaturated fatty acid-induced hyperlipidemia

8/8/2019 Comparative effects of curcumin and photo-irradiated curcumin on alcohol-and polyunsaturated fatty acid-induced

1/8

Pharmacological Research, Vol. 46, No. 3, 2002doi:10.1016/S1043-6618(02)00149-4, available online at http://www.idealibrary.com on

COMPARATIVE EFFECTS OF CURCUMIN AND PHOTO-IRRADIATEDCURCUMIN ON ALCOHOL- AND POLYUNSATURATED

FATTY ACID-INDUCED HYPERLIPIDEMIAR. RUKKUMANI a, M. SRI BALASUBASHINI a , P. VISHWANATHAN b and V.P. MENON a,

a Department of Biochemistry, Faculty of Science, Annamalai University, Annamalai Nagar 608 002,Tamil Nadu, India, b Department of Pathology, Faculty of Medicine, Raja Muthiah Medical College,

Annamalai University, Annamalai Nagar 608 002, Tamil Nadu, India

Accepted 1 July 2002

It is a knownfact that ethanol increases lipid levels inhumansand experimental animals.Reportsshowthat the increased intakeof polyunsaturated fatty acids (PUFAs) along with alcohol produces variouspathological changes in liver resulting in hyperlipidemia. Heating of oil rich in PUFA producesvarious lipid peroxidative end products, which aggravate the pathological changes. In the presentstudy, we have investigated theeffect of curcumin (C)andphoto-irradiated curcumin (IC) on alcohol-and PUFA-induced hyperlipidemia. Our results showed that the activities of alkaline phosphatase(ALP), -glutamyl transferase (GGT) in plasma and levels of cholesterol, triglycerides (TGs) andfree fatty acids (FFAs) in tissues were increased signicantly in both alcohol + raw as well as heatedPUFA groups compared to normal, but decreased signicantly on treatment with curcumin and IC.The IC treatment decreased the levels more signicantly compared to curcumin. The phospholipids(PLs) were increased signicantly in heart and intestine and decreased in liver and kidney in bothalcohol + raw as well as heated PUFA groups. The levels were signicantly decreased in liver andkidney and increased in intestine and heart in both curcumin- and IC-treated groups. But the effectof IC was more pronounced than curcumin. Histopathological observations were also in correlation

with the biochemical parameters. Thus, photo-irradiated curcumin proves itself to be more effectivethan curcumin in treating the above pathological conditions.

2002 Elsevier Science Ltd. All rights reserved.

K ey words : alcohol, PUFA, curcumin, photo-irradiated curcumin, lipids.

INTRODUCTION

Alcohol, a toxic substance in excess causes variousbiochemical, pharmacogenetic and pathological distur-bances. Ethanol produces fatty liver with ultrastructurallesions both in rats and human. It occurs when the in-tracellular redox potential and redox sensitive nutrientmetabolisms are disturbed by alcohol [1]. An excessiveaccumulation of reducing equivalents favours hepaticlipogenesis, decreases hepatic release of lipoproteins, in-creases the mobilisation of peripheral fat, enhances theuptake of circulating lipids and decreases the fatty acidoxidation, andthus increasesthe retentionof lipidsin liver.

The composition of macronutrient is a determinantof nutritional health. Fat is an important dietary com-ponent, which affects both growth and health. Dietary

Correspondingauthor. Departmentof Biochemistry, Facultyof Science,Annamalai University, Annamalai Nagar 608 002, Tamil Nadu, India.E-mail: [email protected]

role of polyunsaturated fatty acids (PUFAs) is gaininga greater attention now a day in the research world. Incontrast to earlier epidemiological studies showing thereduced risk of coronary heart disease (CHD) due toPUFA intake [2], current data on dietary fats indicatethat it is not just the presence of PUFA but the type of PUFA that is important. A high PUFA n-6 content andn-6/n-3 ratio in dietary fats is considered to be athero-genic and diabetogenic. The newer heart friendly oilslike sunower oil possess this undesirable PUFA contentand thus the excess intake of these vegetable oils areactually detrimental to health [3]. Moreover, heating of oil produces various peroxidative changes. During deepfat frying, when the fat is used repeatedly, oxidative andthermal effects result in the formation of many volatileand non-volatile products, some of which are toxic de-pending on the level of intake [4]. Ingestion of decompo-

sition products formed as a result of thermal abuse andoxidation of frying oils is known to lead to a variety of diseases [5]. Thus, the ingestion of alcohol along with

1043-6618/02/$ see front matter 2002 Elsevier Science Ltd. All rights reserved.


2/8

258 Pharmacological Research, Vol. 46, No. 3, 2002

PUFA and heated PUFA is expected to alter the lipidlevels. In our present study, we have investigated the inu-ences of curcumin (bis-4-hydroxy-3-methoxy-phenyl)-1,6-hepta-diene-3,5-dione, an active principle of Curcumalonga and photo-irradiated curcumin on alcohol- and

Fig. 1. (A) Control animals liver: H&E, 20. Hepatocytes arranged in a linear pattern without dilatation of the sinusoids and portal triad ( ).(B) Alcohol + raw PUFA fed rats liver: H&E, 20. Sinusoidal dilatation ( ), congestion ( ), fatty changes especially microvesicular type( ) . (C) Alcohol + raw PUFA + curcumin fed rats liver: H&E, 20. Fatty changes of hepatocyte especially microvesicular ( ) and dilatation ( )of the sinusoids; ( ) indicates portal vein. (D) Alcohol + heated PUFA fed rats liver: H&E, 20. Focal fatty changes of parenchyma, predominantly

macro-vesicular( ).(E) Alcohol + heated PUFA + curcumin fedrats liver: H&E, 20.Fattychanges of hepatocytes especiallymicrovesicular( )and dilatation of the sinusoids ( ). (F) Alcohol + heated PUFA + irradiated curcumin fed rats liver: H&E, 20. Fatty changes of macrovesicular type( ).(G)Controlrats kidney: H&E, 20. Normalglomeruli ( ) and tubules. (H) Alcohol + rawPUFA fedrats kidney:H&E, 20. Cloudyswelling

of tubule ( ) and congested blood vessels ( ). (I) Alcohol + raw PUFA + curcumin fed rats kidney: H&E, 20. Area of haemorrhage ( ). (J)Alcohol + heated PUFA fed rats kidney: H&E, 20. Fatty inltrate ( ).(K)Alcohol + heated PUFA + curcumin fedrats kidney: H&E, 20. Multipleareas of haemorrhage ( ). (L) Alcohol + heated PUFA + irradiated curcumin fed rats kidney: H&E, 20. Multiple areas of haemorrhage ( ).

PUFA-induced hyperlipidemia, since little or no work hasbeen done on treating these conditions andcompared theireffects. Curcumin on UV irradiation produces vanillinand ferulic acid, the stable photoproducts [6], and no dataare available regarding its hypolipidemic effects.


3/8

Pharmacological Research, Vol. 46, No. 3, 2002 259

Fig. 1. (Continued ).

MATERIALS AND METHODS

Female Albino rats, Wistar strain of body weight rang-ing 140150 g bred in the central Animal House, RajahMuthiah Medical College, Chidambaram, were fed on

pellet diet (Agro Corporation Private Limited, Banga-lore, India) and water, ad libitum . The standard pelletdiet comprised 21% protein, 5% lipids, 4% crude -bre, 8% ash, 1% calcium, 0.6% phosphorus, 3.4% glu-cose, 2% vitamins and 55% carbohydrates and providesmetabolisable energy of 3600 kcal kg 1. The animalswere housed in plastic cages under controlled condi-tions of 12-h light/12-h dark cycle, 50% humidity andat 30 3 C.

Materials used Sunower oil: Sunower oil marketed by Gold

Winner was purchased fromlocal market, Chidambaram,Tamil Nadu, India.

Curcumin (C): Curcumin was obtained fromcentral drug house privatelimited, Mumbai, India.

Heated PUFA: Sunower oil was subjected toheating at 180 C for 30min,twice.

Irradiatedcurcumin (IC):

Curcumin was subjected tophoto-oxidation by curcuminexposing curcumin to brightsunlight for 5 h continuously.

Allother chemicalsandsolventsused were of analyticalgrade.

Animals and treatment Group 1: Control rats (rats given standard pellet

diet and glucose solution isocaloric toethanol and a high fat diet, orally usingan intragastric tube).

Group 2: Rats given 20% ethanol [7] orallyusing an intragastric tube and a highfat diet (15%) (raw sunower oilmixed with the diet).

Group 3: Rats given 20% ethanol and a high fatdiet (15%) (thermally oxidisedsunower oil).

Group 4: Rats given curcumin (80 mg kg 1

body weight) [8], dissolved in 20%ethanol (7.99g kg 1 body weight)orally using an intragastric tube and ahigh fat diet (15%) (raw sunower oil).

Group 5: Rats given curcumin (80 mg kg 1

body weight) dissolved in 20% ethanol(7.9gkg 1 body weight) and a high fatdiet (15%) (heated sunower oil).

Group 6: Rats given photo-irradiated curcumin(80 mgkg 1 body weight) dissolved in20% ethanol (7.9g kg 1 body weight)and a high fat diet (15%) (heatedsunower oil) ( Table I ).

At the end of experimental period (45 days), the ratswere sacriced after an overnight fast by decapitation.

Tissues (liver, heart, kidney and intestine) were removed,cleared off blood and immediately transferred to ice-coldcontainers containing 0.9% NaCl for various estimations.


4/8


Table IExperimental design

Group No. of rats Treatment Dose administered

Normal (N) 8 Glucose 36.2 g kg 1 body weight (b.w.)Alcohol + raw PUFA (A + R) 8 20% alcohol + high fat diet 7.9 g kg 1 b.w. ethanol + 15% fat

(raw sunower oil)Alcohol + heated PUFA (A + H) 8 20% alcohol + high fat diet 7.9 g kg 1 b.w. ethanol + 15% fat(heated sunower oil)

Alcohol + raw PUFA + curcumin(A + R + C)

8 20% alcohol + high fat diet + curcumin 7.9g kg 1 b.w. ethanol + 15% fat(raw sunower oil) + 80mgkg 1

b.w. curcuminAlcohol + heated PUFA +

curcumin (A + H + C)8 20% alcohol + high fat diet + curcumin 7.9g kg 1 b.w. ethanol + 15% fat

(heated sunower oil)+ 80mgkg 1 b.w. curcumin

Alcohol + heated PUFA + photo-irradiated curcumin (A + H + IC)

8 20% alcohol + high fat diet+ irradiated curcumin

7.9gkg 1 b.w. ethanol + 15% fat(heated sunower oil) + 80mgkg 1

b.w. irradiated curcumin

Average food intake per day: 15 g/150g rat. Total calories per day: 508 kcal/150 g rat.

For histopathological study, two animals from each groupwere perfused with formalin (10%) and the tissues wereseparated and stored in 10% formalin. They were latersectioned using a microtome, dehydrated in graded alco-hol, embedded in parafn section andstained with haemo-toxylin and eosin (H&E).

Biochemical estimationsThe activities of alkaline phosphatase (ALP) (EC

3.1.3.1)were assayed byPNPP method [9],usingareagentkit, and -glutamyl transferase (GGT) (EC 2.3.2.2) byxed time method of Fiala et al. [10]. The tissue lipidswere extracted according to the method of Folch et al.[11] , cholesterol was estimated by Zlatkis et al. method[12], phospholipids (PLs) by the method of Zilversmitand Davis [13], free fatty acids (FFAs) by Falholt et al.method [14] and triglycerides (TGs) by Foster and Dunn[15] method.

Statistical analysisThedatagiven in thetables areaveragevalues standard

deviation (SD). Data were analysed statistically by anal-ysis of variance (ANOVA) and groups were compared byleast signicant difference (LSD).

RESULTS

Changes in the activities of plasma -glutamyltransferase and alkaline phosphatase

Activities of GGT and ALP are given in Table II . Theactivities of GGT and ALP were increased signicantlyin alcohol + raw and thermally oxidised PUFA groupscompared to normal. Curcumin treatment signicantlydecreased the activities of GGT and ALP compared toabove groups. Treatment of alcohol + thermally oxidised

PUFA with photo-irradiated curcumin showed a signi-cant decrease in the activities of GGT and ALP comparedto treatment with curcumin.

Table IILevels of marker enzymes in plasma (values are mean SD from

six rats in each group)

Groups GGT (IU l 1 ) ALP (IU l 1 )

1. N 0.597 0.06 85.880 7.002. A + R 1.658 0.16 a 174.020 15.85 a

3. A + H 2.508 0.14 a 239.560 22.14 a

4. A + R + C 0.783 0.13 b ,c 119.780 10.21 a ,c

5. A + H + C 1.433 0.12 a ,d 177.410 16.28 a ,d

6. A + H + IC 1.150 0.16 a ,d ,e 149.147 12.13 a ,d , f

F -ratio 160.764 77.259

ANOVA followed by LSD. Groups 26 are compared with group 1;group 4 is compared with group 2; groups 5 and 6 are compared withgroup 3; group 6 is compared with group 5. a P 0.001. b P 0.01.c P 0.001. d P 0.001. e P 0.001. f P 0.01.

Changes in lipidsThe levels of cholesterol ( Table III ), TG (Table IV ) and

FFA ( Table V ) were increased signicantly in alcohol +raw as well as heated PUFA groups compared to normal.But they were decreased signicantly in curcumin-treatedgroup. The decrease was more signicant in IC-treatedgroup compared to curcumin. The levels of PL ( Table VI )were increased in heart and intestine and decreased in

liver and kidney in alcohol + raw as well as heated PUFAgroups. Butcurcumin treatmentdecreasedthePL levels inheart and intestine and increased in liver and kidney. Sim-ilar changes occurred in IC group but more signicantlythan curcumin treatment.

Histopathological changes in liver Histopathological changes of liver are given in Table

VII. The liver samples of alcohol + raw PUFA animalsshowedsinusoidal dilatation, congestion andfattychangesof microvesicular type and occasional macrovesiculartype fatty changes. On treatment with curcumin, the liver

showed dilatation and microvesicular fatty changes. Thealcohol + heated PUFA animals liver showed featherydegeneration of hepatocyte and focal fatty inltrate. On


5/8


Table IIILevels of tissue cholesterol (values are mean SD from six rats in each group)

Groups Liver (mg/100g tissue) Hear t (mg/100g tis sue) Kidney (mg/100g tissue) Intestine (mg/100g tissue)

1. N 335.667 26.36 182.167 17.28 340.000 59.38 174.000 22.592. A + R 464.00 32.42 a 400.000 26.89 a 624.000 39.30 a 426.667 30.22 a

3. A+

H 653.667

34.33a

496.000

26.86a

656.000

48.03a

496.000

44.13a

4. A + R + C 377.667 31.68 b ns 304.000 26.89 a ,b 469.339 39.81 a ,b 336.000 30.37 a ,c

5. A + H + C 517.500 28.28 a ,c 411.000 18.36 a ,c 528.667 26.18 a ,c 414.333 30.31 a ,c

6. A + H + IC 452.667 40.67 a ,c ,d 373.333 16.52 a ,c ,d 464.500 9.03 a ,c ,d 370.00 17.62 a ,c ,e

F -ratio 80.454 134.784 50.139 86.630

ANOVA followed by LSD. Groups 26 are compared with group 1; group 4 is compared with group 2; groups 5 and 6 are compared with group 3;group 6 is compared with group 5; ns: no signicance. a P 0.001. b P 0.001. c P 0.001. d P 0.01. e P 0.05.

Table IVLevels of tissue triglycerides (values are mean SD from six rats in each group)

Groups Liver (mg/100g tissue) Heart (mg/100g tissue) Kidney (mg/100g tissue) Intestine (mg/100g tissue)1. N 332.37 15.89 341.12 22.31 461.38 29.96 371.75 25.822. A + R 542.80 35.85 a 503.75 54.65 a 690.98 34.98 a 524.17 16.67 a

3. A + H 607.89 37.15 a 581.65 55.09 a 795.94 70.59 a 638.50 34.4 a

4. A + R + C 424.73 25.34 a ,b 424.21 33.55 a ,c 546.92 25.25 b ,d 443.89 37.27 a ,b

5. A + H + C 575.01 29.55 a ,e 502.93 25.19 a , f 669.12 42.97 a ,e 542.29 39.29 a ,e

6. A + H + IC 467.13 45.33 a ,e ,g 450.64 26.12 a ,e ,g 591.42 36.39 a ,e ,h 493.39 23.36 a ,e ,h

F -ratio 49.348 27.096 45.862 52.87

ANOVA followed by LSD. Groups 26 are compared with group 1; group 4 is compared with group 2; groups 5 and 6 are compared with group 3;group 6 is compared with group 5. a P 0.001. b P 0.001. c P 0.01. d P 0.01. e P 0.001. f P 0.01. g P 0.05. h P 0.01.

Table VLevels of tissue free fatty acids (values are mean SD from six rats in each group)

Groups Liver (mg/100g tissue) Hear t (mg/100g tis sue) Kidney (mg/100g tis sue) Intestine (mg/100g tissue)

1. N 645.20 63.38 483.55 37.45 386.84 37.45 253.86 39.732. A + R 980.43 71.95 a 770.68 63.10 a 746.42 62.97 a 387.00 36.32 a

3. A + H 1160.53 98.57 a 834.14 85.50 a 894.57 85.60 a 519.82 54.36 a

4. A + R + C 761.80 67.90 b ,c 573.00 11.78 c ,d 483.55 45.31 c ,d 310.18 34.08 b ,e

5. A + H + C 942.49 87.28 a , f 647.26 10.45 a , f 628.03 31.61 a , f 443.62 45.08 a ,g

6. A + H + IC 760.34 74.29 b , f ,h 580.00 14.14 d , f , i 562.09 41.54 a , f , i 360.73 32.65 a , f , j

F -ratio 34.56 47.65 69.10 31.68

ANOVA followed by LSD. Groups 26 are compared with group 1; group 4 is compared with group 2; groups 5 and 6 are compared with group 3;group 6 is compared with group 5. a P 0.001. b P 0.05. c P 0.001. d P 0.01. e P 0.01. f P 0.001. g P 0.01. h P 0.001.i P 0.05. j P 0.01.

Table VILevels of tissue phospholipids (values are mean SD from six rats in each group)

Groups Liver (mg/100g tissue) Hear t (mg/100g tis sue) Kidney (mg/100g tissue) Intestine (mg/100g tissue)

1. N 1708.00 76.52 1051.66 44.00 1500.00 53.67 680.00 48.942. A + R 1040.00 61.97 a 1400.00 61.97 a 920.00 61.97 a 1140.00 65.73 a

3. A + H 883.33 46.33 a 1620.00 65.73 a 800.00 61.97 a 1260.00 29.44 a

4. A + R + C 1390.00 79.75 a ,b 1173.33 41.31 a ,b 1326.66 68.90 a ,c 883.33 29.44 a ,b

5. A + H + C 1240.00 61.97 a ,c 1346.66 78.66 a ,c 1206.67 77.63 a ,c 1060.00 48.99 a ,c

6. A + H + IC 1356.66 48.03 a ,c ,d 1210.83 17.44 a ,c ,e 1298.33 24.88 a ,c , f 965.00 32.09 a ,c ,d

F -ratio 123.325 78.809 114.968 97.655

ANOVA followed by LSD. Groups 26 are compared with group 1; group 4 is compared with group 2; groups 5 and 6 are compared with group 3;group 6 is compared with group 5. a P 0.001. b P 0.001. c P 0.001. d P 0.01. e P 0.05. f P 0.001.


6/8


Table VIIHistopathological changes in liver

Microscopic observations N A+ R A + R + C A + H A + H + C A + H + IC

Nuclear disintegration A ++ + +++ ++ +Sinusoidal congestion A ++ + +++ ++ +Portal inammation A ++ + +++ ++ +Steatosis in zones 2 and 3 A ++ + +++ ++ +Cytoplasmic vacuolation A ++ + +++ ++ +Focal degeneration A ++ + +++ ++ +Fatty changes A ++ + +++ ++ +Micronecrosis A ++ + +++ ++ +Feathery degeneration A ++ + +++ ++ +

A + R PUFAalcohol + raw PUFA; A + R PUFA + Calcohol + raw PUFA + curcumin; A + H PUFAalcohol + heated PUFA; A + HPUFA + Calcohol + heated PUFA + curcumin; A + H PUFA + ICalcohol + heated PUFA + irradiated curcumin. A: absent; + : 05 LPF; ++ :510 LPF; +++ : >10 LPF.

Table VIIIHistopathological changes in kidney

Microscopic observations N A+ R A + R + C A + H A + H + C A + H + IC

Fatty changes A ++ + +++ ++ +Inammation of parenchyma A ++ + +++ ++ +Vessel congestion A ++ + +++ ++ +Haemorrhage A ++ + +++ ++ +

A + R PUFAalcohol + raw PUFA; A + R PUFA + Calcohol + raw PUFA + curcumin; A + H PUFAalcohol + heated PUFA; A + HPUFA + Calcohol + heated PUFA + curcumin; A + H PUFA + ICalcohol + heated PUFA + irradiated curcumin. A: absent; + : 05 LPF; ++ :510 LPF; +++ : >10 LPF.

treatment with curcumin, congestion, dilatation and mi-crovesicular fatty changes were seen. Photo-irradiatedcurcumin-treated animals showed only macrovesiculartype fatty changes ( Fig. 1AF ).

Histopathological changes of kidneyTable VIII gives the histopathological changes of kid-

ney. The kidney samples of alcohol + raw PUFA-treatedanimals showed fatty inltration, cloudy swelling andcongested blood vessels. On treatment with curcumin,the samples showed areas of focal haemorrhage. Hyalinecast, cloudy swelling, fatty inltrate and parenchymalinammation were seen in alcohol + heated PUFA groupanimals. Treatment with curcumin showed multiple ar-eas of haemorrhage, cloudy swelling and fatty inltrate.Treatment with photo-irradiated curcumin showed onlymultiple areas of haemorrhage ( Fig. 1GL ).

DISCUSSION

Marked alterations in lipid metabolism have been re-ported in chronic ethanol feeding [16]. The main pathwayof alcohol degradation, alcohol dehydrogenase pathwayleads to increased NADH synthesis. This striking redoxchange inhibits TCA cycle, fatty acid oxidation, lipopro-tein export and increases fatty acid uptake [17] and thuspredisposing fatty liver.

Ethanol treatments to rats are known to cause centrilob-

ular necrosis in the liver leading to theaccumulationof fat.The fats from peripheral adipose tissues are translocatedto liver, brain and kidney for accumulation. Antonenkov

et al. [18] have reported that chronic ethanol ingestion re-sults in moderate hypercholesterolemia and hypertriglyc-eridemia and increased concentration of lipids in liver.

The increased cholesterol may be due to increased

-hydroxy-methyl-glutaryl CoA (HMG CoA) reductaseactivity by ethanol, which is the rate-limiting step incholesterol biosynthesis [19]. Reports show that the dietrich in PUFA stimulates the production of chylomicronsby the intestine [20], this may be the reason for the in-creased levels of cholesterol in alcohol + raw PUFAgroup. Moreover, reports show that higher plasma choles-terol levels are observed in heated oil fed group [21] andso in alcohol + heated PUFA groups cholesterol levelsare higher.

Fielding et al. [22] have found the increased plasma TGconcentrations after acute ethanol ingestion. In vitro alco-hol has been shown to exert a direct effect on myocardiallipid metabolism, causing an increase in fatty acid esteri-cationanda decrease in oxidation, thus, accumulatingTG.The increased TG levels after PUFA ingestion may be dueto the increased availability of substrate, FFA for esteri-cation. Since the availability of FFAs is more in heatedPUFA group, the TG levels are also comparatively higher.

Phospholipids are the vital components of biomem-brane. They are the primary targets of peroxidation andcanbe altered byethanol [23]. The decrease in the levelsof phospholipids in liver and kidney may be due to increasedactivity of phospholipases in these tissues. It hasalso beensuggested that the decreased levels of phospholipids are

due to increased degradation of muscle phospholipids.Earlier studies have demonstrated that chronic exposureto ethanol may lead to progressive increase in membrane


7/8


phospholipase A 2 activity. Jaya et al. [24] have reporteda decrease in the phospholipid content in liver and kidneyof alcohol fed rats. The increased levels in other tissuesmay be due to the increased availability of FFA. SincePUFA is a component of PL, the increased PUFA intake

may increase the levels of PL in other tissues.The free fatty acid levels are enhanced in alcohol fedgroup, which may be attributed to increased formationof acetate, which in turn forms FFA. The increasedNADH/NAD + ratio also favours fatty acid synthesis.Thus, the levels of FFA are increased in liver, heart,kidney and intestine. Increased FFA in alcohol + PUFA(raw + heated) may be due to the increased lipid break-down and increased dietary PUFA, may also eventuallyincrease the FFA levels.

Treatment of curcumin reduces the levels of lipids.Hypocholesterolemic effect of curcumin is due to theincreased HDL formation, which transports the excesscholesterol from extrahepatic tissues to liver where it iscatabolised [25]. Curcumin also decreases the absorp-tion of cholesterol [26]. Reports suggest that curcuminincreases 7 -hydroxylase activities; the main enzymeinvolved in the conversion of cholesterol to bile acid andthus facilitates biliary cholesterol excretion [27].

The exact mechanism by which curcumin lowers otherlipid levels are not known, however, studies have shownthat some of thespices play a vital role in lipidmetabolism,dueto their active principle. Thespices areknown to affectbile acid excretion and thereby inuence lipid levels. Thedecreasedlevels of phospholipids andTG mayalso be due

to the decreased FFA synthesis by curcumin, which maysuppress the enzymes involved in FFA synthesis.The levels of lipids were signicantly reduced in

photo-irradiated curcumin treatment groups comparedto curcumin. Vanillin and ferulic acid, the componentsof IC, due to their effective antioxidant property, protectmembrane and prevent FFA release. Being a substratefor other lipids, FFA decrease may reect on the levelsof lipids. Recent reports show that -oryzanol, a mixtureof ferulic acid can lower cholesterol level in blood andlower the incidence of coronary heart disease. In Japan, ithas been used as a natural antioxidant in foods, beveragesand cosmetics [28]. Thus, the vanillin and ferulic acideffectively reduce lipids by an unknown mechanism.

Fatty changes are the commonest single feature of al-coholic liver disease. Moreover, the ingestion of alcoholalong with raw PUFA induces cytochrome P450 andcauses severe damage to liver. Thus, fatty changes of micro and macrovesicular type, sinusoidal dilatation andcongestion were seen in alcohol + raw PUFA group. Theadministrationof powerful antioxidant curcumin, reducedthe extent of liver damage and so only dilatation andmicrovesicular fatty changes were seen in alcohol + rawPUFA + curcumin group.

During alcohol ingestion, fatty vacuolation will become

severe such that every hepatocyte contains a globule of fat. The source of the hepatic fat depends on the amountof fat in the diet. Thus, heated PUFA further potentiates

the effect. Histological features of alcoholic hepatitis in-clude degenerative changes in the hepatocytes, inltrationof polymorphonuclear leukocytes and deposition of alco-holic hyaline. The alcohol + heated PUFA group, thus,showed focal fatty inltrate and feathery degeneration.

Treatment with curcumin reduced these adverse effects,showed congestion, dilatation and fatty inltration. Thetreatment with photo-irradiated curcumin showed onlymacrovesicular fattychanges, thus,proving itself more ef-fective than curcumin.

Alcohol fedrats normally show fatty inltration,inam-mation of parenchyma and vessel congestion in kidney.Studies have shown that chronic ethanol consumption re-sults in increased ethanol oxidation by kidney, leading tothe formation of reactive oxygen species. Alcohol + rawPUFA groupshowedfatty inltration, cloudy swellingandcongested blood vessels, which may be due to the alco-holic effect further induced by PUFA. Only areas of focalhaemorrhage wereseenin alcohol + rawPUFA + curcumingroup due to the protective role of curcumin.

Alcohol + heated PUFA group showed hyaline castwithin dilated tubule, cloudy swelling, fatty inltrationand parenchymal inammation, which may be attributedto the severe toxicity of alcohol + heated PUFA. Thehyaline may act as an antigen and produce inamma-tory response. Curcumin being an effective antioxidant,decreases the severity. Focal fatty inltration, cloudyswelling and multiple areas of haemorrhage were seen incurcumin treated group and only multiple areas of haem-orrhage were seen in photo-irradiated curcumin-treated

group, thus, proving the efcacy of the later.Thus, our results indicate that IC is more effective thancurcumin in controlling the lipid levels and maintainingthe histology.

REFERENCES

1. Lieber CS, Davidson CS. Some metabolic effects by ethyl alcohol. Am J Med 1962; 33: 31927.

2. Nordey A, Goodnight SH. Dietary lipids and thrombosis, relation-shipsto atherosclerosis: a review. Arteriosclerosis 1990; 10: 14963.

3. Sircar S, Kansra V. Choice of cooking oilsmyths and realities. J Ind Med Assoc 1998; 96: 3047.

4. Alexander JC. Chemical and biological properties related to toxicityof heated fats. J Toxicol Environ Health 1981; 7: 12538.

5. Twex H, Ismail HM, Sumars S. The effect of intermittent heatingon some chemical parameter of rened oils used in Egypt: a publichealth nutrition concern. Int J Food Sci Nutr 1998; 49: 33942.

6. Dahl TA, Bilski P, Reszka K, Chignell CF. Photocytotoxicity of curcumin. Photochem Photobiol 1994; 59: 2904.

7. Rajakrishnan V, Vishwanathan P, Menon VP. Hepatotoxic effect of alcohol on female rats and siblings: effects of n-acetyl cysteine. Hepatol Res 1997; 9: 3750.

8. Rajakrishnan V, Jayadeep A, Arun OS, Sudhakaran PR, Menon VP.Changes in the prostaglandin level in alcohol toxicity: effect of cur-cumin and n-acetyl cysteine. J Nutr Biochem 2000; 11: 50914.

9. King EJ, Armstrong AR. Calcium, phosphorus and phosphatase. In:Practical clinical biochemistry . New Delhi: CBS Publishers, 1988:458.

10. Fiala S, Fiala AE, Dixon B. Gamma glutamyl transpeptidasein chemically induced rat hepatomas and spontaneous mousehepatomas. J Natl Cancer Inst 1972; 48: 1393409.


8/8


11. Folch J, Lees M, Solane SGH. A simple method for isolation andpurication of total lipids from animal tissues. J Biol Chem 1957;26: 497509.

12. Zlatkis A,Zak B,BoylAJ. A new methodfor thedirectdeterminationof serum cholesterol. J Lab Clin Med 1953; 45: 486.

13. Zilversmit DB, Davis AK. Microdetermination of plasma phospho-lipids by trichloroacetic acid precipitation. J Lab Clin Med 1950;35: 155.

14. Falholt K, Falholt W, Lund B. An easy colorimetric method forroutine determination of free fatty acids in plasma. Clin Chim Acta1973; 46: 105.

15. Foster CS, Dunn O. Stable reagents for determination of serumtriglycerides by a colorimetric Hantzsch condensation method. ClinChem Acta 1973; 19: 33840.

16. Day CP, James OF, Brown AS, Bennet MK, Flemming IN, Yera-man SJ. The activity of the metabolic form of hepatic phosphatidatephosphohydrolase correlateswith the severity of alcoholic fatty liverin human beings. Hepatology 1993; 18: 8328.

17. GalliA, Price D, Crabb D. Highlevel expressionof ratclassI alcoholdehydrogenase is sufcient for ethanol induced fat accumulation intransduced HeLa cells. Hepatology 1999; 29: 116470.

18. Antonenkov VD, Popova SV, Panchenkov LF. Inuence of ethanoland debrate on the activity of lipid catabolism, enzymic systems inthe rat liver. Farmalcol Toksikol 1983; 46: 8690.

19. Ashakumari L, Vijayammal PL. Additive effect of alcoholand nicotine on lipid metabolism in rats. Ind J Exp Biol 1993; 31:2704.

20. HocquetteJF, Bauchart D. Intestinal absorption, blood transport andhepatic and muscle metabolism of fatty acids in pre ruminant andruminant animals. Reprod Nut Dev 1999; 39: 2748.

21. Narasimhamurthy K, Raina PL. Long term feeding effects of heatedand fried oils on lipids and lipoproteins in rats. Mol Cell Biochem1999; 195: 14353.

22. Fielding BA, Reids G, Grandy M, Humphreys SM, Evans K, FraynKN. Ethanol with a mixed meal increases postprandial triglyceridesbut decreases postprandial non esteried fatty acid concentrations. Br J Nutr 2000; 83: 597604.

23. Yamada S, Mark KM, Lieber CS. Chronic ethanol consumption al-ters ratliver plasmamembrane andpotentiatesthe release of alkalinephosphatase. Gastroenterology 1985; 88: 1799806.

24. Jaya DS,AugustineJ, Menon VP. Protectiverole of n-acetyl cysteineagainst alcohol andparacetamolinduced toxicity. Ind J Clin Biochem1994; 9: 6471.

25. Soudamini KK, Unnikrishnan MC, Soni KB, KuttanR. Inhibition of lipid peroxidation and cholesterol levels in mice by curcumin. Ind J Physiol Pharmacol 1992; 36: 23943.

26. Rao DS, Chandrasekhara N, Sathyanarayana MN, Srinivasan M.Effect of curcumin on serum and liver cholesterol levels in rat. J Nutr 1970; 100: 130715.

27. BabuPS, SrinivasanK. Hypolipidemic action of curcumin, theactiveprinciple of turmeric ( Curcuma longa ) in streptozotocin induceddiabetic rats. Mol Cell Biochem 1997; 166: 16975.

28. Scavariello EM, Ardlano DB. -Oryzanol: an important componentin rice bran oil. Arch Latinoam Nutr 1998; 48: 712.

Documents

Comparative effects of curcumin and photo-irradiated curcumin on alcohol-and polyunsaturated fatty acid-induced hyperlipidemia