EFFECT OF RIGLA(Portulaca oleracea L.)GRINDED SEEDSON OBESE FEMALE RATS

doi:10.21608/ejas.2020.102640

EFFECT OF RIGLA(Portulaca oleracea L.)GRINDED SEEDSON OBESE FEMALE RATS

Egyptian Journal of Applied Science

Article 7, Volume 35, Issue 3 - Serial Number 1, March and April 2020, Page 39-48

Document Type: Original Article

DOI: 10.21608/ejas.2020.102640

View on SCiNiTO

Abstract

ABSTRACT
Rigla(Portulaca oleracea L.) is a plant used widely for its medicinal properties, many studies confirmed its role as antioxidant and antibacterial, antitumor activities, rich source of vitamins, minerals and omega-3. The aim of the present study was to investigate the effect of Riglagrinded seeds (RGS) on body weight, serum lipid profile, leptin, liver and kidney functions of female obese rats. Thirty adult female albino rats (Sprague-Dawley strain), weighing about (200±10g) were divided into two main groups, the first main group (-ve control=6 rats) was fed on basal diet. The second main group (n=24 rats) fed on high fat diet for four weak, then divided into 4 subgroups from group 2 to group 5. Group 2 (+ve control) fed on high fat diet only. Groups from 3 to 5 fed on high fat diet supplemented with 5%, 10% and 15% grinded seeds of Rigla, respectively. At the end of the experiment period (4 weeks), rats were scarified and serum was collected for biochemical analyzes. Results indicated that supplementation with Rigla caused signification decreased (p

Highlights

تأثیر بذور الرجلة المطحونة على اناث الفئران المصابة بالسمنة

جمیلة حرب ، أحمد أمین ، هانی المصری

قسم التغذیة وعلوم الأطعمة ، کلیة الاقتصاد المنزلی

جامعة حلوان ، القاهرة ، مصر

الرجله نبات یستخدم على نطاق واسع لخصائصه الطبیة. تؤکد العدید من الدراسات دورها کمضاد للأکسدة ومضاد للبکتیریا ، وأنشطة مضادة للورم ، ومصدر غنی بالفیتامینات والمعادن وأومیجا 3. کان الهدف من الدراسة الحالیة هو بحث تأثیر بذور الرجله المطحونة على وزن الجسم ، ومستوی الدهون فی الدم ، هرمون اللبتین ، ووظائف الکبد والکلى لدى الفئران البدینة. تم تقسیم ثلاثین أنثى من الفئران البیضاء (Sprague-Dawley) ، والتی تزن حوالی (200 ± 10 جم) إلى مجموعتین رئیسیتین ، تم تغذیة المجموعة الرئیسیه الأولى (الضابطه السالبه 6 فئران) على النظام الغذائی الأساسی. المجموعة الثانیة (ن = 24 فأر) تتغذى على نظام غذائی عالی الدهون لأربعة أسابیع ، ثم تنقسم إلى 4 مجموعات فرعیة من المجموعة 2 إلى المجموعة 5. المجموعة 2 (المجموعه الضابطه الموجبه) تتغذى على نظام غذائی عالی الدهون فقط. مجموعة من 3 إلى 5 تم تغذیتها على نظام غذائی غنی بالدهون 5٪ و 10٪ و 15٪ من بذور الرجله المطحونة على التوالی. فی نهایة فترة التجربة (4 أسابیع) ، تم ذبح الفئران وجمع السیرم لتحالیل البیوکمیائیه. أوضحت النتائج أن إضافه الرجله للغذاء الأساسی کانت لها تاثیر نقص معنویة (p<0.05) على وزن الجسم للجرذان البدینة مقارنة بالمجموعة الإیجابیة الضابطة. علاوة على ذلک ، انخفض نسبه الجلوکوز ، اللبتین ، الکولیسترول ، الدهون الثلاثیة ، LDL-c ، VLDL-c ، الیوریا ، حمض الیوریک ، الکریاتینین ، انزیمات الکبدAST ، ALT ، ALP بشکل معنوی (p<0.05) فی جمیع المجموعات المعالجة مقارنه بالمجموعه الضابطه الموجبه ، بینما کانت قیم HDL-c فی السیرم مرتفعه معنویاً (p<0.05) مقارنة بالمجموعة الإیجابیة ولذا یمکن التوصیة بأن تستخدم الرجله فى علاج للسمنة.

Keywords

Key Words: Rigla (portulacaoleracea L.); obesity; rats; Lipid profil; liver function; kidney function

Full Text

Egypt. J. of Appl. Sci., 35 (3) 2020 39-48

EFFECT OF RIGLA(Portulaca oleracea L.)GRINDEDSEEDSON OBESE FEMALE RATS

Gamela G. Harb ; Ahmed A. Ameen and Hany G. El-Masry

Nutrition and Food Science Department, Faculty of Home Economics, Helwan University, Cairo, Egypt

Key Words: Rigla (portulacaoleracea L.), obesity, rats,Lipid profil , liver function, kidney function

ABSTRACT

Rigla(Portulaca oleracea L.) is a plant used widely for its medicinal properties, many studies confirmed its role as antioxidant and antibacterial, antitumor activities, rich source of vitamins, minerals and omega-3. The aim of the present study was to investigate the effect of Riglagrinded seeds (RGS) on body weight, serum lipid profile, leptin, liver and kidney functions of female obese rats. Thirty adult female albino rats (Sprague-Dawley strain), weighing about (200±10g) were divided into two main groups, the first main group (-ve control=6 rats) was fed on basal diet. The second main group (n=24 rats) fed on high fat diet for four weak, then divided into 4 subgroups from group 2 to group 5. Group 2 (+ve control) fed on high fat diet only. Groups from 3 to 5 fed on high fat diet supplemented with 5%, 10% and 15% grinded seeds of Rigla, respectively. At the end of the experiment period (4 weeks), rats were scarified and serum was collected for biochemical analyzes. Results indicated that supplementation with Rigla caused signification decreased (p<0.05) in body weight of obese ratsin groups 3-5 compared to control positive group. Moreover, serum glucose, leptin, cholesterol, triglycerides, LDL-c, VLDL-c, urea, uric acid, creatinine, AST, ALT, ALP were significantly decreased (p<0.05) in all treated groups, while serum HDL-c was significantly increased (p<0.05) as compared to control positive group. It could be recommended that Rigla may be suitable for obesity.

INTRODUCTION

Obesity, one of the worldwide health problems, it is considered increase in fat mass accumulation and body weight gain.This illness results from the rise of high caloric food intake and reducing physical activity. The main plan for prevention and treatment of obesity is lifestyle change, containing dieting control and physical exercise (Chandrasekaran et al., 2012).

Obesity is the chief risk factor for metabolic syndrome, which is connected with increased risk of several types of chronic diseases, for example hypertension, hyperlipidemia, cardiovascular diseases , fatty liver and insulin resistance. As reported by world health organization (WHO) at least 2.8 million adults die yearly because of being overweight (Al-Faris, 2014).

40Egypt. J. of Appl. Sci., 35 (3) 2020

Herbal medicines can be certain as raw or extracted products distant from plants, and they are commonly used for prevention and treatment of many chronic diseases, such as obesity, because they have fewer side effects compared with pharmacological drugs (Park et al., 2011).

Rigla (Portulaca oleracea L.) is a warm-climate, herbaceous juicy yearly plant. Rigla is an excellent source of omega- 3 fatty acids. Omega-3 fatty acids play an important role in the development of immune function and prevention and treatment of hypertension, coronary artery disease, cancer, and additional inflammatory and autoimmune illnesses It has also sufficiently of antioxidant compounds for example alpha-tocopherol, ascorbic acid and glutathione. Rigla also a good source of coenzyme (Elkhayat et al., 2008 and Park, 2011). There are seven different flavonoids are present in Rigla, such as kaempferol, myricetin, luteolin, apigenin, quercetin, genistein, and genistingenistin (Zhu et al., 2010).

The aim of the present study conducted to evaluate the effect of Riglagrinded seeds on body weight, serum lipid profile, leptin, liver and kidney function of female obese rats.

MATERIALS AND METHODS

Materials:

Grinded seeds of Rigla was obtained from Agriculture Research center, Cairo, Egypt. Casein, cellulose, sucrose, choline chloride, D-L methionine, chemical kits, vitamins and minerals constituents were purchased from El-Gomhoriya Pharm., Cairo, Egypt. Starch, corn oil, beef tallow and sucrose were obtained from Egyptian local market. Thirty adult male albino rats (Sprague-Dawley strain), weighing about (200±10 g) were obtained from the Farm of Experimental Animals, Helwan, Egypt.

Methods:
1. Induction of obesity:

Rats were fed on high fat diet (HFD) containing (saturated fat 19%, soybean oil 1% to provide essential fatty acids, sucrose 10%, casein 20%, cellulose 5%, vitamins mixture 1%, salt mixture 3.5%, choline chloride 0.25% and the remainder is corn starch) for four weeks to induce obesity in rats (Min et al., 2004).

Diet composition and experimental animal design:

The basal diet was formulated according to AIN-93M diet (Reeves et al., 1993). Animals (30 rats) were housed in well conditions in Biological Studies Laboratory of Faculty of Home Economics. Rats were left for seven days as adaptation period and they were allowed to feed standard laboratory food and water. After the period of adaptation, animals were divided into two main groups, as follows: the first group (6 rats) was fed on basal diet and served as a negative control group (-Ve), the second group (24 rats) was fed on HFD (Min et al., 2004). After four weeks, this group was divided equally to four subgroups as follows:-

Egypt. J. of Appl. Sci., 35 (3) 2020 41

Subgroup (1):

Obese rats (positive control group), animals were fed on high fat diet only (HFD).

Subgroup (2):

Obese rats were fed on basal diet supplemented with the grinded seeds of Rigla 5 % from high fat diet (HFD).

Subgroup (3):

Obese rats were fed on basal diet supplemented with the grinded seeds of Rigla 10 % from high fat diet (HFD).

Subgroup (4):

Obese rats were fed on basal diet supplemented with the grinded seeds of Rigla 15 % from high fat diet (HFD).

At the end of the experimental period (4 weeks), rats were fasted overnight before scarifying and blood samples were collected from each rat and were centrifuged at 3000 rpm for 15 min to obtain the serum for biochemical analysis.

Biological evaluation:

Feed intake (FI), body weight gain percent (BWG%) and feed efficiency ratios (FER) were determined according to Chapman et al., (1959).

Biochemical analysis of serum:

Serum glucose and leptin were determined according to the methods described by Trinder (1969) and Zhang et al., (1995),respectively. Aspartate aminotransaminase and alanine aminotransaminase were determined according to Young, (2001), and alkaline phosphates was determined according to Roy, (1970). Serum urea and creatinine were assayed according to Young, (2001),uric acid was dertermined according to Milena, (2003). Serum total cholesterol, triglyceride, high density lipoprotein cholesterol was determined according to Richmond, (1973); Wahlefeld, (1974) and Albers et al., (1983),respectively. Regarding to serum low density lipoprotein cholesterol and very low density lipoprotein cholesterol were calculated according to Fridewald et al., (1972).

42Egypt. J. of Appl. Sci., 35 (3) 2020

Statistical analysis:

Results were expressed as the mean ± standard error (SE). Data were statistically analyzed for variance “ANOVA” test at P ≤ (0.05) using SPSS statistical software, version 20 was used for these calculations (Armitage and Berry, 1987).

RESULTS AND DISCUSSION

Result illustrated in Table (1) showed the effect of diet supplemented with Rigla seeds powder(RSB) on body weight gain (BWG), peritoneal fat(PF), feed intake (FI) and feed efficiency ratio (FER) of obese rats. Data revealed that BWG, peritoneal fat and FER significantly (P< 0.05) increased in the positive control group (+ve) compared to negative control group (-ve). Whereas, all treated group significantly (P<0.05) decreased compared to positive control group (+ve), while FI decreased in +ve control compared to -ve control but all treated group increased (P< 0.05) compared to positive control group. The best result for FER, weight loss and peritoneal fat pad was the group that treated with the level of 15 % Rigla powder.

Table (1): Effect of Rigla (Portulaca oleracea L.) GrindedSeedson Initial Body Weight (IBW), Final Body Weight(FBW), Body Weight Gain(BWG), Peritoneal Fat(PF), Feed Intake(FI), and Feed Efficiency Ratio(FER) of Obese Rats

Groups	IBW	FBW	BWG	PF	FI	FER
Groups	(g)	(g)	%	%	(g/day/rat)	FER
G1: Control (-ve)	235.4±1.91^a	252.4±1.69^bc	7.22±0.30^c	3.31±0.20^b	16	0.038±0.001^b
G2: Control (+ve)	235.6±1.39^a	266.2±1.20^a	12.99±0.60^a	7.08 ±0.13^a	15	0.073±0.003^a
G3:5% RGS	237.6±1.24^a	258.4±1.69^b	8.75±0.21^b	2.96±0.10^bc	16.6	0.044±0.001^b
G4:10% RGS	240.6±1.20^a	253.0±1.30^bc	5.15±0.21^d	2.66±0.07^c	17.3	0.025±0.001^c
G5:15% RGS	241.2±1.77^a	248.8±2.08^c	3.15±0.20^e	1.96±0.11^d	17.7	0.015±0.001^d

*Mean values are expressed as means ± SE.

*Mean values at the same column with the same superscript letters are not statistically significant at P<0.05.

The results agreed with Hussein, (2010) who found the obese rats fed on high fat diet with additional Rigla (150 and 300 mg/kg body weight) for 8 weeks, significantly inhibited body weight gain, blood glucose, triglyceride, total cholesterol, LDL-c, HDL-c and free fatty acids. Mohamed, (2011) reported thatrats feed on 5 g of Rigla seeds twice daily significantly reduced in body weight.

Egypt. J. of Appl. Sci., 35 (3) 2020 43

The concentration of serum leptin and glucose were recorded in Table (2), results indicated that leptin and glucose level in the positive control group significantly increased (P<0.05) compared to negative control group. While all treated groups with Rigla decreased (P<0.05) compared to the positive control group. It was observed the highest improvement for leptin and glucose level was seen at the group fed on 15% of Rigla.

Table (2): Effect of RiglaGrindedSeedson Serum Leptin and Glucose of Obese Rats

Groups	Leptin	Glucose
Groups	µg/L	mg/dL
G1: Control (-ve)	1.79±0.09^d	70.4±0.75^d
G2: Control (+ve)	5.14±0.22^a	114.4±1.69^a
G3:5% RGS	4.06±0.14^b	95.6±1.16^b
G4:10% RGS	3.45±0.19^bc	84.4±1.63^c
G5:15% RGS	2.83±0.10^c	81.0±1.04^c

*Mean values are expressed as means ± SE.

*Mean values at the same column with the same superscript letters are not statistically significant at P<0.05.

The obtained result was in agreement with Bahceciet al., (1999) reportedthat high fat diet results in an increasing body fat proportion with hypertriglyceridemia, hypercholsetrolemia and hyperleptinemia. Stachon et al., (2006) expressed positive relation between plasma leptin levels and epididymal fat mass, liver and heart weights.Also, the result agreed with Bendong, (2012) found that Rigla reduced the leptin hormone in obese rats. Morever Rigla seedsshowed a significant reduction in serum levels of blood glucose and insulin (Mohamed, 2011).

Data in Table (3) showed the effect of Rigla at different levels on lipid profile in rats fed on high fat diet. Results revealed that serum TG, TC, LDL-c and VLDL-c were significant (P<0.05) increased in positive control group compared to negative control group, all treated groups with Rigla significant (P<0.05) decrease in TG, TC, LDL-c and VLDL-c compared to positive control group. Regarding HDL significantly decreased in positive control group compared to negative control group, while all treated groups with Rigla significantly increased in HDL compared to positive control group. The highest improvement for lipid profile was recorded at the group fed on 15% of Rigla.

Ali et al., (2014) found that TC, LDL-c, and TG exposed statistically significant changes over time (one month) in the Rigla group (P<0.05) exposed that the changes in the stated parameters were statistically just for LDL-c and TG, while others did not differ significantly. P. oleracea L. may have positive effects on serum lipids profile which may be attributed to its polyphenolic and antioxidant compounds. This herbal drug seems to be well-tolerated in adolescent population as well. Further studies are recommended.

Also, Niharika andSukumar, (2016) proved that extract of Rigla was able to reduce TGs, TC, LDL-c, VLDL-c and increase in HDL-c.

44Egypt. J. of Appl. Sci., 35 (3) 2020

Table (3): Effect of RiglaGrindedSeeds on lipid profile of Obese Rats

Groups	TG	TC	HDL-c	LDL-c	VHDL-c
Groups	mg/dl
G1:Control (-ve)	77.34±0.84^e	89.06±0.51^d	57.31±0.56^a	16.29±0.93^d	15.46±0.16^e
G2:Control (+ve)	133.29±0.84^a	141.60±0.96^a	40.30±0.56^d	74.65±1.19^a	26.65±0.17^a
G3:5% RGS	101.55±1.27^b	109.10±1.27^b	44.96±0.57^c	43.83±0.87^b	20.31±0.24^b
G4:10% RGS	96.15±0.83^c	94.58±0.39^c	48.45±0.46^b	26.90±0.56^c	19.23±0.16^c
G5:15% RGS	83.40±0.85^d	91.76±0.93^cd	50.62±0.57^b	24.46±1.53^c	16.68±0.17^d

*Mean values are expressed as means ± SE.

*Mean values at the same column with the same superscript letters are not statistically significant at P<0.05.

The concentration of urea, uric acid and creatinine recorded in Table (4), in the positive control group showed significantly increase compared to negative control group, while all treated groups with Rigla decreased compared to the positive control group. Whereas, the highest improvement for urea, uric acid and creatinine at the group fed on 15% of Rigla.

Table (4): Effect of RiglaGrindedSeedson Urea, Uric Acid and Creatinine of Obese Rats

Groups	Urea	Uric Acid	Creatinine
Groups	mg/dl
G1: Control (-ve)	25.2±0.37^d	3.38±0.20^c	0.60±0.01^c
G2:Control (+ve)	63.2±1.95^a	6.37±0.18^a	2.23±0.08^a
G3:5% RGS	57.0±0.70^ab	5.26±0.10^b	0.77±0.01^b
G4:10% RGS	47.4±0.50^bc	4.81±0.10^b	0.71±0.01^bc
G5:15% RGS	42.4±0.81^c	3.96±0.13^c	0.65±0.01^bc

*Mean values are expressed as means ± SE. *Mean values at the same column with the same superscript letters are not statistically significant at P<0.05.

Karimi et al., (2010) indicated that treatment with extracts of Rigla in the highest dose (0.8 and 2 g/kg), reduced blood urea nitrogen and serum creatinine. Tubular necrotic damage was not observed and Rigla extract may protect renal toxicity and might to limit renal injury.

Result in Table (5) showed the serum concentration of AST, ALT and ALP were significantly increased (P<0.05) in the positive control group compared to the negative control group, while significantly (P<0.05) decreased in all groups treated with RSB was observed.

Theresults in agreement with Ausha, (2011) who testedthe effect ofRigla on rats fed on high fat diet. Rigla reduced levels of liver enzymes hypoglycemic, hypolipidemic, effect of reducing liver enzymes AST and ALT and improve liver function. Also, Mohamed, (2011)found that the powder of Rigla seed decreased the serum levels of liver ALT and AST.

Egypt. J. of Appl. Sci., 35 (3) 2020 45

Based on the results of the present study, it could be concluded that Rigla supplementation at the tested groups may provide beneficial effects in body weight reduction as well as improving lipid profil and liver and kidney functions in obese rats.

Table (5): Effect of RiglaGrindedSeedson liver function of Obese Rats

Groups	AST	ALT	ALP
Groups	(µ/L)
G1: Control (-ve)	57.0±0.70^e	23.2±0.58^d	51.51±1.99^c
G2: Control (+ve)	92.2±1.28^a	38.0±0.70^a	81.16±2.14^a
G3:5% RGS	84.8±1.20^b	31.6±0.50^b	71.19±2.95^b
G4:10% RGS	78.6±0.50^c	26.2±0.86^c	66.63±2.11^b
G5:15% RGS	69.2±0.86^d	24.8±0.58^cd	60.65±2.15^b

*Mean values are expressed as means ± SE.

*Mean values at the same column with the same superscript letters are not statistically significant at P<0.05.

REFERENCES

Albers, N. ;V. Benderson and G. Warnick (1983): Enzaymatic determination of high density lipoprotein cholesterol, Selected Methods. Clin. Chem., 10:91-99.

Al-Faris, N. A. (2014): Biological evaluation of the weight reduction capacity of selected herbal products available in the Kingdom of Saudi Arabia.Nutrition and Food Sciences J,3(1): 7-13.

Ali, M. ; K. Roya and J. Hadi (2014): The effect of portulaca oleracea on obesity.med Arch Jun,68:198-199.

Armitage, G.Y. and W.G. Berry (1987): Statistical methods 7^th Ed. Ames., Iowa State University. Press. 39-63.

Ausha, M. ; M. Venkateswariu andV. Prabhakaran (2011):Hepatoprotective activity of aqueous extract of Portulaca oleracea in combination with lycopene in rats.Indian J Pharmacol. , 43: 563–567.

Bahceci, M. ; A. Tuzcu ; M. Akkus ;M. Yaldiz and A. Ozbay (1999):The effect of high fat diet on the development of obesity and serum leptin level in rats.Eat weight Disorder,4(3):128-132.

Bendong, C. (2012):Effects of aqueous extract of Portulaca oleracea L. on oxidative stress and liver, spleen leptin, PARα and FAS mRNA expression in high-fat diet induced mice.Molecular Biology Reports, 39:7981–7988.

46Egypt. J. of Appl. Sci., 35 (3) 2020

Chandrasekaran, C. ;M. Vijayalakshmi ; K. Prakash ; V.Bansal ; J. Meenakshi and A. Amit (2012): Herbal approach for obesity management. Am J Plant Sci, 3: 1003-1014.

Chapman, D.G. ;R. Gastilla and J. A. Campbell (1959): Evaluation of protein in foods: 1- A Method for the determination of protein efficiency ratio. Can. J. Biochem. Phys., 37:679- 86.

Elkhayat, E.S. S.R. Ibrahim and A.M. Aziz (2008): Portulen a new diterpene from PortulacaoleraceaL.”Journal of Asian Natural ProductsResearch, 10:1039–1043.

Fridewald, W. ;R. Leve and D. Fredrickson (1972): Estimation of the concentration of low densitylipoprotein separated by three different methods. Clin. Chem., 18:499-502.

Hussein, M. (2010): Purslane extract effects on obesity-induced diabetic rats fed a high-fat diet. Malaysian Journal of Nutrition, 16:419-429.

Karimi, G. ; A. Khoei ; A. Omidi ; M. Kalantari ;J. Babaei and B. Taghiabadi (2010): Protective effect of aqueous and ethanolic extracts of Portulaca oleracea against cisplatin induced nephrotoxicity. Iranian Journal of Basic Medical Sciences, 13:31-35.

Milena, J. (2003): Predrag Djurdjevic and DejanStankov .J.SCS., 68(8-9): 691-698.

Min, L. ;S. Ling ;L. Yin ;C.W. Stephen ;J. Randy ;D. David and T. Patrick (2004): Obesity induced by a high-fat diet downregulates apolipoprotein A-IV gene expression in rat hypothalamus. Am. J. Physiol. Endocrinol Metab., 287: 366-370.

Mohamed, K. (2011): Effect of portulaca oleracea seed in treatment of type-2diabetes mellitus pation as adjunactive and altractive theraps.journal of Ethanopharmacology, 137:643-651.

Niharika, S. and D. Sukmar (2016): Hypolipidemic effect of portulaca oleracea in rats fet on high cholesterol diet. J nutrition and food science, 30:120-125.

Park, K. (2011): Textbook of preventive and social medicine. 21thed. Jabalapur: Bhanot Press: 234-239.

Park,J. ;J. Kim ;M. Park and J. Yun (2011): Potential agents for cancer and obesity treatment with herbal medicines from the green garden. J Plant Sci, 16: 1065-1073.

Egypt. J. of Appl. Sci., 35 (3) 2020 47

Reeves, P. ;F. Nielsen and G. Fahmy (1993): AIN-93.Purified diets for laboratory rodents: Final reports of the American Institute of Nutrition adhoe wriling committee of reformulation of the AIN-76 A Rodent Diet. J. Nutr., 123:1939-51.

Richmond, N. (1973): Colorimetric determination of total cholesterol and high density lipoprotein cholesterol (HDL-c). Clin. Chem., 19: 1350- 1356.

Roy, S. (1970): colorimetric method of serum alkaline phosphatase. Journal of Clinical Chemistry, 16:431-432.

Stachon, M. ;E. Furstenberg andE. Gromadzka, (2006): Effects of high fat diet on body composition ,hypothalamus NPYand plasma leptin corticosterone levels in rats. J Endocer, 30:69-74.

Trinder, P. (1969): Determination of blood glucose using 4-aminophenazone. J. Clin. Path., 22: 246.

Wahlefeld, A. (1974): Methods of Enzymatic Analysis. Academic Press, Chapter, 5: 1831-1835.

Young, D. (2001): Effect of disease on clinical lab Tests, 4^th ed. AACC press.

Zhang, Y.; R. Proenca ; M. Maffei ; M. Barone ; L. Leopold and J.M. Friedman, (1995): positional cloning of the mouse obese gene and its human homologue, [erratum 374:479]. Nature., 72:425-432.

Zhu,H. ;Y. Wang ; Y.Liu ;I. Xiaand T.Tang (2010):Analysis of flavonoids in Portulacaoleracea L. by UV-vis Spectrophotometry with comparative study on different extraction technologies,” Food Analytical Methods, 3:90–97.

تأثیر بذور الرجلة المطحونة على اناث الفئران المصابة بالسمنة

جمیلة حرب ، أحمد أمین ، هانی المصری

قسم التغذیة وعلوم الأطعمة ، کلیة الاقتصاد المنزلی

جامعة حلوان ، القاهرة ، مصر

48Egypt. J. of Appl. Sci., 35 (3) 2020

References