Anti-diabetic and hypolipidaemic properties of garlic (Allium sativum) in streptozotocin-induced diabetic rats

Transcription

1 Int J Dietes & Metolism (27) 15: Anti-dietic nd hypolipidemic properties of grlic (Allium stivum) in streptozotocin-induced dietic rts Mrth Thomson, Zin M. Al-Amin, Khled K. Al-Qttn, Lemi H. Shn nd Muslim Ali Deprtment of Biologicl Sciences, Fculty of Science, Kuwit University,, Kuwit. Astrct In this study the hypoglycemic, hypocholesterolemic nd hypotriglyceridemic effects of grlic were studied in streptozotocin (STZ)-induced dietic rts. Compred to norml (non-dietic) rts, STZ-induced dietic rts hd pproximtely 2% higher serum glucose, 5% higher serum cholesterol nd 3% higher serum triglyceride levels s well s 86% higher urinry protein levels. Dily tretment of STZ-induced dietic rts with n extrct of rw grlic (5mg/kg intrperitonelly) for seven weeks significntly lowered serum glucose, cholesterol nd triglyceride levels. Compred to control dietic rts, grlic-treted rts hd 57% less serum glucose, 4% lower serum cholesterol levels nd 35% lower triglyceride. In ddition, urinry protein levels in grlic-treted dietic nimls were 5% lower compred to the dietic controls. In contrst, the incresed urine output nd wter intke of dietic rts were not ffected y grlic tretment. These results indicte tht rw grlic possesses eneficil potentil in reversing proteinuri in ddition to reducing lood sugr, cholesterol nd triglycerides in dietic rts. Therefore, grlic could e of gret vlue in mnging the effects nd complictions of dietes in ffected individuls. Key words: Dietes, Grlic, hypoglycemic ctivity, hypolipidemic ctivity, proteinuri Introduction Grlic (Allium stivum) is one of the most populr hers used worldwide to reduce vrious risk fctors ssocited with crdiovsculr diseses. Grlic, memer of the Lilicee fmily, is common food for flvour nd spice nd it is one of the hers most commonly used in modern folkloric medicine. Grlic ws n importnt medicine to the ncient Egyptins s listed in the medicl text Codex Eers (c. 155 BC) especilly for the working clss involved in hevy lour ecuse it ws n effective remedy for mny liments such s hert prolems, hedche, ites, worms nd tumours. Grlic is stted to possess mny therpeutic enefits. Grlic s strong odour is lrgely due to sulphur-contining compounds (e.g. S-llylcysteine sulphoxide), which re elieved to ccount for most of its medicinl properties. 1 Actully, grlic contins vriety of effective compounds tht exhiit nticogulnt (nti-thromotic), 2,3,4,5,6 ntioxidnt, 7,8 ntiiotic, 9,1,11 hypocholesterolemic, 12 hypoglycemic, 1 s well s hypotensive ctivities As mentioned ove, lthough lrge numer of sulphur- Received on: 16/3/27 Accepted on: 18/1/27 Correspondence to: Prof. Muslim Ali, Dept of Biologicl Sciences, Fculty of Science, Kuwit University, P.O. Box 5969, 136-Sft, Kuwit. Tel.: ; Fx: ; emil: limuslim@hotmil.com thiosulphintes re present in sufficient quntities t norml consumption levels (3-5 g per dy). Allicin hs een shown to e importnt in mny helth effects of grlic. 14 However, the nti-cncer effect of grlic might e shred etween llicin nd other unidentified compounds. 15 Grlic contins out 1% lliin, which is converted enzymticlly y llicinse to llicin, nd other sulphur-contining compounds. 16 Grlic hs een found to e effective in lowering serum glucose levels in STZ-induced s well s lloxn-induced dietic rts nd mice. Most of the studies showed tht grlic cn reduce lood glucose levels in dietic mice, rts nd rits. 14 Augusti nd Sheel consistently showed tht S-llyl cysteine sulphoxide, (llicin), sulphur-contining mino cid in grlic (2 mg/kg ody weight), hd potentil to reduce the dietic condition in rts lmost to the sme extent s did glienclmide nd insulin Aged grlic extrct ws lso effective in preventing drenl hypertrophy, hyperglycemi nd elevtion of corticosterone in mice mde hyperglycemic y immoiliztion stress. 19 In ddition, Liu nd co-workers reported tht oth grlic oil nd dillyl trisulphide improved glycemic control in STZ-induced dietic rts. 2 Ingestion of grlic juice resulted in etter utiliztion of glucose in glucose tolernce tests performed in rits, while llicin t dose of 25 mg/kg ws 6% s effective s tolutmide in lloxn-induced dietic rits. 21 In contrst, grlic powder intke (6.25% y weight in diet) for 12 dys reduced hyperphgi nd polydipsi, ut did not 18

2 Thomson et l lter either hyperglycemi or hypoinsulinemi in STZinduced dietic mice. 22 Similrly, Bluchnejdmojrd nd Rohgni found no hypoglycemic effect of n queous extrct of grlic in rts with STZ-induced dietes lthough they did oserve significnt effect of grlic on vsculr rectivity Liu nd co-workers hve speculted tht these inconsistent results re t lest prtly due to the use of different preprtions or derivtives of grlic in the different studies. 2 St nd coworkers hve estlished tht the chemicls present in grlic product re lrgely dependent on the processing conditions, such s temperture, durtion of preprtion, nd extrction solvents used. 25 In humns, the hypoglycemic effect of grlic is not well documented. Most reports hve shown significnt effect of grlic on lood glucose of norml helthy individuls ut not in dietic ptients. Thus the role of grlic in dietes tretment/prevention in humns is yet to e confirmed. 13 The im of the present study ws to investigte the efficcy of n queous extrct of rw grlic in controlling serum glucose, cholesterol, triglyceride nd urine protein levels in STZ-induced dietic rts treted dily intrperitonelly (IP) for period of 7 weeks. Since there hve een vrile reports out the use of different preprtions of grlic, s discussed ove, n queous extrct of rw grlic ws used in the present study. Use of this preprtion is lso 4-5, 26, 27,28 consistent with our previous work with grlic. Mterils nd Methods Extrct preprtion Aqueous grlic extrct ws prepred from loclly ville grlic uls. The grlic uls were peeled on crushed ice. Then 5 g of the peeled grlic ws cut into smll pieces nd homogenized in 7 ml of cold, sterile.9% NCl in the presence of some crushed ice. The homogeniztion ws crried out in lender t high speed using 3 second ursts for totl of 1 minutes. The homogenized mixture ws filtered 3 times through cheesecloth, the filtrte ws centrifuged t 2 RCF for 1 minutes nd the cler superntnt ws diluted to 1 ml with norml sline. The concentrtion of this grlic preprtion ws considered to e 5 mg/ml on the sis of the weight of the strting mteril (5 g/1 ml). The queous extrct of grlic ws stored in smll liquots t -2 C until use. The stility of the preprtion during storge hs een previously estlished in pltelet ggregtion studies (unpulished oservtions). Tretment of Dietic Rts Mle Sprgue-Dwley rts weighing g (prents purchsed from Lortory Animls Inc., Englnd) nd mintined on norml diet nd filtered tp wter d liitum were used in the experiment. For seline dt, lood ws drwn from ll nimls y crdic puncture under ether nesthesi nd llowed to clot. Immeditely, the clotted lood ws centrifuged t 35 RPM for 3 minutes. The serum ws seprted nd stored t 8 o C for lter nlysis. The nimls were rndomly divided into helthy group (8 rts) nd streptozotocin (STZ)-treted group (initilly 2 rts). The STZ-treted rts were injected IP with 6 mg streptozotocin/kg ody weight in volume of.5 ml sline following n overnight fst. 29 After period of three dys, lood ws drwn from the fsting STZ-treted nimls nd serum ws prepred nd stored for lter nlysis s descried ove. Serum glucose levels were determined immeditely nd the STZ-treted rts determined to e dietic due to high serum glucose level (>35 mg/dl) were rndomly divided into two groups contining 8 nimls ech: Group 1, the control dietic group, ws injected IP dily with sline for the tretment period, nd Group 2, the grlic-treted group, ws injected IP dily with 5 mg/kg of the grlic extrct. After periods of two, five nd seven weeks, lood ws drwn from the fsting rts y crdic puncture, nd serum ws prepred nd stored for lter nlysis s descried ove. Rts were weighed efore the strt of the experiment nd then weekly during the experimentl period. Animls were monitored for generl helth during the tretment period. Twenty-four hour wter intke ws mesured dily. Twenty-four hour urine output ws mesured in ech tretment group efore STZ dministrtion nd t two, five nd seven weeks y housing the nimls in metolic cges nd collecting the urine in clirted 25 ml ttched continer. The collected urine ws centrifuged t 3 RPM for 1 min to remove ny prticulte mteril. Urine smples were then stored t -8 C for protein determintion. At the end of the experiment, the rts were scrificed under sodium pentoritone nesthesi ccording to the guidelines for euthnsi in the Guide for the Cre nd Use of Lortory Animls. 3 Assys Serum glucose, cholesterol nd triglyceride were determined spectrophotometriclly using kits supplied y CARO Co., Germny. Urinry protein ws determined y the Coomssie Blue dye inding method of Brdford. 31 Sttisticl Anlysis The dt re expressed s men ± SEM. Redings within group were compred using the one-wy ANOVA nlysis nd redings etween groups were compred using the independent smple test. Sttisticl nlysis ws performed using SPSS (Version 14). A level of p <.5 ws considered to e significnt. Results Figure 1 shows chnges in the serum glucose levels in STZinduced dietic rts in response to 5 mg/kg grlic extrct dministrtion. It is cler from the dt tht the serum glucose levels of the control dietic nimls continued to increse during the 7 weeks of the experiment compred to the post-stz injection level. In contrst, the grlic-treted dietic rts showed significntly reduced serum glucose levels during the tretment period when compred to the control dietic rts. At weeks 2, 5 nd 7 of grlic extrct tretment, the serum glucose levels of the grlic-treted dietic rts were reduced y 29%, 68% nd 57%, respectively in comprison to control dietic rts. 19

3 Anti-dietic effects of grlic 8 7 control dietic grlic treted dietic norml lood glucose (mg/dl) c c c 1 post-stz 2 wks 5 wks 7 wks Figure 1: Serum glucose levels in STZ-induced dietic rts treted with queous extrct of grlic. Glucose levels were mesured in serum of norml rts, STZ-induced dietic rts (control dietic) nd grlic-treted STZ-induced dietic rts (grlic-treted dietic). Norml glucose levels were verged over the experimentl period nd re depicted s roken line. Anlysis ws done fter STZ injection (post-stz), nd fter 2, 5 nd 7 weeks of tretment. : Significntly incresed compred to norml (p<.5), : Significntly decresed compred to control dietic (p<.5). c: Significntly decresed compred to post-stz (p<.5) weight % with weeks during the experiment norml control dietic grlic treted dietic pre-stz post-stz Figure 2: Weights of norml nd STZ-induced dietic rts over the grlic tretment period. Weights were mesured in norml rts, STZ-induced dietic rts (control dietic) nd grlic-treted STZ-induced dietic rts (grlic treted dietic). The nimls were weighed efore STZ injection (pre-stz), one week fter STZ-injection (post-stz), nd then weekly during the tretment period of 7 weeks. Weights re plotted s percentiles with the strting weights ll stndrdized to 1%. : Significntly decresed compred to norml t week 7 (p<.5), Significnt difference etween control dietic nd grlic treted dietic rts (p<.5). However, grlic tretment did not reduce serum glucose of dietic nimls to norml levels. Norml rts gined weight significntly throughout the experimentl period, while oth the control dietic nd grlic-treted dietic nimls hd significntly lower ody weights when compred to norml nimls (Figure 2). However, grlic-treted dietic rts mintined their initil weights during the 7-week tretment period lthough t the end of the experiment their ody weights were significntly less thn those of norml rts. In contrst, the control dietic rts showed significnt weight loss when compred to oth the norml rts nd the grlic-treted dietic rts t the end of the 7-week experiment. 11

4 Thomson et l control dietic grlic treted dietic norml Blood cholesterol (mg/dl) post-stz 2 wks 5 wks 7 wks Figure 3: Serum cholesterol levels in STZ-induced dietic rts treted with queous extrct of grlic. Cholesterol levels were mesured in serum of norml rts, STZ-induced dietic rts (control dietic) nd grlic-treted STZ-induced dietic rts (grlic treted dietic). Norml cholesterol levels were verged over the experimentl period nd re depicted s roken line. Anlysis ws done fter STZ injection (post STZ), nd fter 2, 5 nd 7 weeks of tretment. Significntly incresed compred to norml (p <.5), Significntly decresed compred to dietic control (p <.5). 1 9 control dietic grlic treted dietic norml lood triglycerides (mg/dl) post-stz 2 wks 5 wks 7 wks Figure 4: Serum triglyceride levels in STZ-induced dietic rts treted with queous extrct of grlic. Triglyceride levels were mesured in serum of norml rts, STZ-induced dietic rts (control dietic) nd grlic-treted STZ-induced dietic rts (grlic treted dietic). Norml triglyceride levels were verged over the experimentl period nd re depicted s roken line. Anlysis ws done fter STZ injection (post STZ), nd fter 2, 5 nd 7 weeks of tretment. : Significntly incresed compred to norml (p<.5), : Significntly decresed compred to dietic control (p<.5). In Figure 3, one week fter STZ-injection (post-stz), the serum cholesterol levels of the dietic rts were significntly higher thn the norml levels. After tretment, the serum cholesterol levels of grlic-treted dietic rts were significntly lower in comprison with the control dietic rts. The serum cholesterol reduction elicited y grlic ws sustined throughout the course of tretment with serum cholesterol levels elow the norml level fter 5 nd 7 weeks of grlic tretment. Figure 4 shows reduction in serum triglyceride levels to norml levels in grlic-treted dietic rts through the 7 weeks of tretment. In contrst, oth cholesterol nd triglyceride levels remined elevted in the control dietic nimls throughout the experimentl period. 111

5 Anti-dietic effects of grlic urine protein (mg/24 hrs) control dietic grlic treted dietic norml post-stz 2 wks 5 wks 7 wks Figure 5: Urinry protein levels in STZ-induced dietic rts treted with queous extrct of grlic. Protein levels were mesured in urine of norml rts, STZ-induced dietic rts (control dietic) nd grlic-treted STZ-induced dietic rts (grlic treted dietic). Norml urinry protein levels were verged over the experimentl period nd re depicted s roken line. Anlysis ws done fter STZ injection (post STZ), nd fter 2, 5 nd 7 weeks of tretment. Significntly incresed compred to norml (p<.5). Significntly decresed compred to control dietic (p<.5). urine output (ml/dy) control dietic grlic treted dietic norml control dietic post- 2 wks 5 wks 7 wks Figure 6: Urine output in STZ-induced dietic rts treted with queous extrct of grlic. Urine output ws mesured in rts fter STZ-injection (post STZ) nd fter 2, 5 nd 7 weeks of grlic tretment of STZ-induced dietic rts. Norml urine output levels were verged over the experimentl period nd re depicted s roken line. Urine output levels in control dietic rts were verged over the experimentl period nd re presented s n open r. Significntly incresed compred to norml (p<.5). The urinry protein levels of grlic-treted dietic rts were significntly lower thn urinry protein levels in the control dietic rts (Figure 5). One week fter STZinjection (post-stz), the urinry protein level of the control dietic rts ws doule tht of the norml rts nd remined elevted throughout the experimentl period. In contrst, t weeks 2, 5 nd 7 of grlic tretment, urinry protein levels in grlic-treted dietic rts were lowered y 28%, 53% nd 49%, respectively, with the urinry protein levels t 5 nd 7 weeks of grlic tretment reching the norml level. In the nlysis of the dt collected on urine output, it ws found tht the effect of grlic tretment ws insignificnt (Figure 6). In fct, the urine output of the grlic-treted dietic rts remined elevted during the 7 weeks of tretment. In contrst, the grlic-treted dietic rts hd decresed wter intke fter 2 weeks of tretment, fter which 112

6 Thomson et l 3 control dietic grlic treted dietic norml 25 wter intke (ml/dy) control dietic post-stz 2 wks 5 wks 7 wks Figure 7: Wter intke in STZ-induced dietic rts treted with queous extrct of grlic. Wter intke ws mesured in rts fter STZ-injection (post STZ) nd fter 2, 5 nd 7 weeks of grlic tretment of STZ-induced dietic rts. Norml wter intke levels were verged over the experimentl period nd re depicted s roken line. Wter intke levels in control dietic rts were verged over the experimentl period nd re presented s n open r. Significntly incresed compred to norml (p<.5). Significntly decresed compred to control dietic (p<.5). there ws no significnt reduction in wter intke in the grlic-treted dietic nimls compred to the control dietic rts (Figure 7). Discussion Dietes mellitus is the most common endocrine disorder tht ffects more thn 194 million people worldwide. If nothing is done to control this disese, the numer will exceed 333 million y 225 (6.3% of popultion). In 23, Kuwit ws mong the five countries of the world with the highest dietes prevlence in the dult popultion (12.8%). 32 In ddition to the primry effects of dietes, dietes is ccompnied y incresed risk fctors such s hyperglycemi, dyslipidemi, hypertension, decresed firinolytic ctivity, incresed pltelet ggregtion, nd severe therosclerosis Mny synthetic drugs hve een developed for the tretment of dietes. However, these drugs hve limits in terms of efficcy nd side effects. 35 Therefore, there is much interest in discovering nturl tretments without negtive side effects tht cn reduce these risk fctors in dietic ptients. Grlic hs een reported to possess vriety of medicinl properties including hypoglycemic, hypocholesterolemic nd hypolipidemic ctivities. 36 However, previous studies on the hypoglycemic ctivity of grlic preprtions hve produced vrile results. 7,17-24 Since we re interested in the eneficil effects of consumption of whole grlic, we chose to study the complete queous extrct. In ddition, in our previous studies, we hve otined consistent results using n queous extrct of grlic in oth rts nd 4,5,6,7,8,9,1,11,12, 26, 27,28 rits. In these experiments, the queous extrct of grlic ws given IP since, in previous experiments, oth orl nd IP dministrtions were found to e eneficil nd IP dministrtion required less hndling of the nimls. 6 The dosge ws chosen to e 5 mg/kg since it is sfe mount 14, 36 of grlic to e given dily nd does not cuse toxicity. In ddition, we hve previously dministered this dose (5 mg/kg) vi the IP route with no detrimentl effects in terms 26, 28 of toxicity nd mode of dministrtion. The effects of the extrcts of rw grlic were oserved over period of seven weeks. STZ-induced dietic rts showed significnt elevtion of serum glucose, cholesterol nd triglyceride levels. Our results confirmed tht rw grlic hs significnt hypoglycemic, hypocholesterolemic nd hypolipidemic effects. Therefore, the present study reinforces the findings of previous ppers tht grlic hd 17,18,19,2,21, 37, 38 significnt effect in reducing lood glucose, cholesterol 12 nd triglyceride levels 8 in dietic niml models. We lso oserved tht the weight loss tht occurred in STZinduced dietic rts ws ttenuted y grlic tretment. In ddition, t 7 weeks, the grlic-treted dietic group hd lower mortlity (one out of eight) compred to the control dietic group (three out of eight). These chnges my e reflection of the improved helth of the grlic-treted dietic nimls. Proteinuri is mjor predictor of glomerulr injury nd elevted rtes of protein excretion re selective mrkers of progressive nephropthy. 39 STZ-induced dietic rts re chrcterized y the development of proteinuri. 4 Furthermore, it hs een shown tht STZ hs no long-term 113

7 Anti-dietic effects of grlic direct effects on the kidney, ut STZ hs secondry effects on the kidney s result of the development of dietes mellitus. 41 Our results showed tht rw grlic my llevite renl dmge cused y STZ-induced dietes, which ws mnifested y the significnt lowering of urinry protein levels in the rw grlic-treted rts. Similr results were oserved in lloxn-induced dietic rts y El-Demerdsh et l. who emphsized the lleviting effect of grlic on renl dmge. 42 However, grlic tretment did not llevite the chrcteristic polydypsi or polyuri oserved in the STZ-induced dietic rts. This is in greement with the results of Liu et l. who lso oserved no chnge in wter intke or urine output in STZ-induced dietic rts treted with grlic oil or dillyl trisulphide. 2 It is not cler how grlic ctully works in lleviting hyperglycemi. The hypoglycemic ction of grlic could possily e due to n increse in pncretic secretion of insulin from β-cells, relese of ound insulin or enhncement of insulin sensitivity. It hs een previously suggested tht grlic (llicin) cn enhnce serum insulin y effectively comining with compounds like cysteine, which would spre insulin from SH group rections which re common cuse of insulin inctivtion. 21 Another mechnism proposed y Augusti nd Sheel sttes tht the ntioxidnt effect of S-llyl cysteine sulfoxide, n isolted product from grlic, my contriute to its eneficil effect in dietes. 7 Jin nd Vys postulted tht grlic my ct s n ntidietic gent y incresing either the pncretic secretion of insulin from the β-cells or relese of ound insulin. 43 This explntion is supported y the results of Liu nd coworkers who reported tht tretment of STZ-induced dietic rts with grlic oil or dillyl sulphide resulted in incresed serum insulin levels. 2 The results of this study strongly suggest tht grlic my e very useful in the llevition of dietic complictions s well s in the prevention of the development of therosclerosis nd nephropthy generlly oserved in dietic ptients. In the future, further work is needed to investigte the ctive ingredients in grlic. In ddition, more prmeters should e studied such s insulin enhncement, HA 1c level, free rdicl production, etc., to elucidte the mechnism of ction of the ctive constituents of grlic. Acknowledgments This work ws supported y Kuwit University grnt numer SB1/99 nd the College of Grdute Studies, Kuwit University for which the uthors re grteful. References 1. Augusti KT. Therpeutic vlues of onion (Allium cep L.) nd grlic (Allium stivum L.). Indin J Exp Biol 1996; 34: Kiesewetter H, Jung F, Pindur G, et l. Effect of grlic on thromocyte ggregtion, microcircultion, nd other risk fctors. Int Clin Phrmcol Ther Toxicol 1991; 29: Ali M, Thomson M. Consumption of grlic clove dy could e eneficil in preventing thromosis. Prostglndins Leukot Essent Ftty Acids 1995; 53: Bordi T, Mohmmed N, Thomson M, Ali M. An evlution of grlic nd onion s ntithromotic gents. Prostglndins Leukot Essent Ftty Acids 1996; 54: Ali M, Thomson M, Alnqee MA, et l. Antithromotic ctivity of grlic: its inhiition of the synthesis of thromoxne-txb 2 during infusion of rchidonic cid nd collgen in rits. Prostglndins Leukot Essent Ftty Acids 199; 41: Thomson M, Mustf T, Ali M. Thromoxne-B 2 levels in serum of rits receiving single intrvenous dose of queous extrct of grlic nd onion. Prostglndins Leukot Essent Ftty Acids 2; 63: Augusti KT, Sheel CG. Antiperoxide effect of S-llyl cysteine sulfoxide, insulin secretgogue, in dietic rts. Experienti 1996; 52: Anwr MM, Meki AR. Oxidtive stress in streptozotocin-induced dietic rts: effects of grlic oil nd meltonin. Comp Biochem Physiol A Mol Integr Physiol 23; 135: Bkri IM, Dougls CW. Inhiitory effect of grlic extrct on orl cteri. Arch Orl Biol 25; 5: Rees LP, Minney SF, Plummer NT, Slter JH, Skyrme DA. A quntittive ssessment of the ntimicroil ctivity of grlic (Allium stivum.) World J Microiol Biotechnol 1993; 9: Yoshid H, Iwt N, Krsuzki H, et l. Antimicroil ctivity of compound isolted from n oil-mcerted grlic extrct. Biosci Biotechnol Biochem 1998; 62: Ali M, Al-Qttn KK, Al-Enezi F, Khnfer RM, Mustf T. Effect of llicin from grlic powder on serum lipids nd lood pressure in rts fed with high cholesterol diet. Prostglndins Leukot Essent Ftty Acids 2; 62: Bnerjee SK, Mulik SK. Effect of grlic on crdiovsculr disorders: review. Nutr J 22; 1: Jmison JR. Grlic (Allium stivum). In: Clinicl Guide to Nutrition nd Dietry Supplements in Disese Mngement. London: Churchill Livingstone, 23: Hssn HT: Ajoene (nturl grlic compound): new nti-leukemi gent for AML therpy. Leuk Res 24; 28: Block E, Ahmd S, Ctlfmo JL, Jin MK, Apitz- Cstro R. Antithromotic orgnosulfur compounds from grlic, structurl, mechnistic nd synthetic studies. J Am Chem Soc 1986; 18: Sheel CG, Augusti KT. Antidietic effects of S-llyl cysteine sulphoxide isolted from grlic Allium stivum Linn. Indin J Exp Biol 1992; 3: Sheel CG, Kumud K, Augusti KT. Anti-dietic effects of onion nd grlic sulfoxide mino cids in rts. Plnt Med 1995; 61: Ksug S, Ushijim M, Morihr N, et l. Effect of ged grlic extrct (AGE) on hyperglycemi induced y immoiliztion stress in mice. Nippon Ykurigku Zsshi 1999; 1999:

8 Thomson et l 2. Liu C-T, Hse H, Lii C-K, Chen P-S, Sheen L-Y. Effects of grlic oil nd dillyl trisulfide on glycemic control in dietic rts. Eur J Phrmcol 25; 516: Mthew PT, Augusti KT. Studies on the effect of llicin (dillyl disulphide-oxide) on lloxn dietes I. Hypoglycemic ction nd enhncement of serum insulin effect nd glycogen synthesis. Indin J Biochem Biophys 1973; 1: Swnston-Fltt SK, Dy C, Biley CJ, Fltt PR. Trditionl plnt tretments for dietes. Studies in norml nd streptozotocin dietic mice. Dietologi 199; 33: Bluchnejdmojrd T, Rohgni M. Endotheliumdependent nd -independent effect of queous extrct of grlic on vsculr rectivity on dietic rts. Filoterpi 23; 74: Bluchnejdmojrd T, Rohgni M. Grlic extrct ttenutes time-dependent chnges in the rectivity of isolted ort in streptozotocin-dietic rts. Life Sci 23; 73: St EJ, Lsh L, St JE. A commentry on the effects of grlic extrction nd formultion on product composition. J Nutr 21; 131: 1118S-1119S. 26. Alnqee M, Thomson M, Bordi T, Ali M. Histopthologicl effects of grlic on liver nd lung of rts. Toxicol Lett 1996; 85: Al-Qttn KK, Alnqee MA, Ali M. The ntihypertensive effect of grlic (Allium stivum) in the rt two kidney-one clip Goldltt model. J Ethnophrmcol 1999; 66: Al-Qttn KK, Khn I, Alnqee MA, Ali M. Mechnism of grlic (Allium stivum) induced reduction of hypertension in 2K-1C rts. A possile medition of N/H exchnger isoform-1. Prostglndins Leukot Essent Ftty Acids 23; 69: Giorgino F, Chen JH, Smith RJ. Chnges in tyrosine phosphoryltion of insulin receptors nd 17, moleculr weight non-receptor protein in vivo in skeletl muscle in streptozotocin-induced dietic rts: effects of insulin nd glucose. Endocrinology 1992; 13: Veterinry Medicl Cre (Chpter 3). In: Guide for the Cre nd Use of Lortory Animls. Wshington DC: Institute for Lortory Animl Reserch, Americn Acdemy of Sciences, Brdford MM. A rpid nd sensitive method for the quntifiction of microgrm quntities of protein utilizing the principle of protein-dye inding. Anl Biochem 1976; 72: Dietes Atls, Second Edition. Belgium: Interntionl Dietes Federtion (IDF), Reusch JE. Dietes, microvsculr complictions, nd crdiovsculr complictions: wht is it out glucose? J Clin Invest 23; 112: Willims G, Pickup JC. Hndook of Dietes, Third Edition. Mlden MA: Blckwell Pulishing Co., Rng HP, Dle MM, Ritter JM, Moore PK. Phrmcology, 5th Edition. Edinurgh: Churchill Livingstone, Elsevier Science Ltd., 23: McKenn DJ, Jones K, Hughes K, Humphrey S: Botnicl Medicines, the desk reference for mjor herl supplements, Second Edition. New York: The Hworth Herl Press, 22: Eisenrth GS, Kotzin BL. Enumerting utorective T cells in peripherl lood: ig step in dietes prediction. J Clin Invest 23; 111: Jelodr GA, Mleki M, Motdyen MH, Sirus S. Effect of fenugreek, onion nd grlic on lood glucose nd histopthology of pncres of lloxn-induced dietic rts. Indin J Med Sci 25; 59: Kene WF, Eknoyn G. Proteinuri, luminuri, risk, ssessment, detection, elimintion (PARADE): A position pper of the ntionl kidney foundtion. Am J Kidney Dis 1999; 33: Greive KA, Osick TM, Russo LM, Comper WD. Nephrotic-like proteinuri in experimentl dietes. Am J Nephrol 23; 23: Rsch R. Prevention of dietic glomerulopthy in streptozotocin dietic rts y insulin tretment. Kidney size nd glomerulr volume. Dietologi 1979; 16: El-Demerdsh FM, Yousef MI, El-Ng NI. Biochemicl study on the hypoglycemic effects of onion nd grlic in lloxn-induced dietic rts. Food Chem Toxicol 25; 43: Jin RC, Vys CR. Hypoglycemic ction of onion nd grlic. Am J Clin Nutr 1975; 28: