Received on : 10-10-2017
Accpected on : 01-12-2017
Address for correspondence
Dr. Vishnu Kumar
Department of Biochemistry,
Era's Lucknow Medical College & Hospital, Lucknow-226003 Email:madhwapur1976@gmail.com Contact No.+91-8953589756

Original Article

ERA’S JOURNAL OF MEDICAL RESEARCH

VOL.4 NO.2

EFFECTS OF NATURAL PRODUCTS ON BODY WEIGHT AND BIOCHEMICAL

PARAMETERS IN HEALTHY RATS

Vishnu Kumar, Farzana Mahdi, Jitendra Kumar Saxena*, Raj Kumar Singh**,

Mrinal Ranjan Srivastava*** and Sharique Ahmad****

*Department of Biochemistry, Central Drug Research Institute, Lucknow, U.P., India-226003,

**Department of Biochemistry, TSM Medical College, Opposite Amausi Railway Station, Lucknow, U.P., India-226008,

***Department of Community Medicine, **** Department of Pathology, Department of Biochemistry

Era's Lucknow Medical College & Hospital, Sarfarazganj, Lucknow, U. P., India-226003

ABSTRACT

The present study was undertaken to evaluate effect of natural products i.e.

Anthocephalus indicus; KADAM, roots, Hibiscus rosa sinensis roots,

Tinospora cordifolia stem and Cassia tora seeds in normal healthy rats. In

this study ethanol extract of above mentioned medicinal plants had macerated with aqueous gum acacia (2%, w/v) suspension and fed orally (500 mg/kg bw p.o.) to male adult healthy normal rats of Charles Foster strain for 30 days. Results of this study showing that alcoholic extracts caused no any signicant reduction in blood glucose, total cholesterol, triglyceride, phospholipids, free fatty acid, lipid peroxide and no signicant increased in post heparin lipolytic activity, but on the other hand as per pre- existing data and my published studies in diabetic patients and diabetic experimental animals showed that extracts exerting all above effects signicantly. That's why it is very clear here if healthy person will take

natural products, it never cause hypoglycemia, hypolipidemia and under weight. Natural products also not cause any signicant change on hepato-specic parameters. Thus from this study we conclude that, natural products are safe, non toxic and free from side effects, in comparison to synthetic drugs.

KEYWORDS: Natural Products, Hepato-specic parameters, Post heparin lipolytic activity, plant antioxidant.

INTRODUCTION

During Vedic period Aryans compiled their work related to herbal remedy in holy Vedas when they came to north India. References about a number of herbal remedies have been mentioned in 'Rig-Veda' (about 200 B.C.). In 'Atharva-Veda' (about 200 B.C.) description of medicinal plants has been made under separate chapter 'Ayurveda'. It was Charak (about 600 B.C.) who made the scientic classication of herbal drugs on remedial properties in his renowned treatise 'Charak Samhita' (A compendium of general medicine). Anthocephalus indicus; KADAM, Hibiscus rosa

sinensis, Tinospora cordifolia and Cassia tora are some of them. (1-3)

Design of newer, safer and effective chemotherapeutic agents is one-way of addressing this situation. This in turn enforces the need to develop protocols to evaluate the efcacy of herbal drugs. (4)

But effects of above Anthocephalus indicus; KADAM,

Hibiscus rosa sinensis, Tinospora cordifolia and Cassia tora in healthy rats had not studied in detail. In view of the above considerations the present study was designed to investigate the adverse effects of

Anthocephalus indicus, Hibiscus rosa sinenss, C. tora

and T. cordifolia in normal healthy rats.

ANTHOCEPHALUS INDICUS

Anthocephalus cadamba Miq., Syn. A. indicus, (Family-Rubiaceae) commonly called kadamba enjoys a hallowed position in Ayurveda - an Indian indigenous system of medicine. It is also named as Kadam. (5-10)

 

 

 

 

 

 

Tree of A. indicus

 

Flowers of A. indicus

 

 

 

 

 

Fruits of A. indicus

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EFFECTS OF NATURAL PRODUCTS ON BODY WEIGHT AND BIOCHEMICAL PARAMETERS IN HEALTHY RATS

HIBISCUSROSASINENSIS:

TheherbHibiscusrosa-sinensisLinn[Malvaceae

]has antidiabetic and so many benecial effects. (11-15)

Flower of Hibiscus

Roots of Hibiscus

TINOSPORACORDIFOLIA

India is bestowed with enormous biodiversity of medicinal plants. Among them Tinospora cordifolia has a wide array of bioactive principles as well as it has been proven medicinally important plant, have not received considerable scientic attention. (16-18)

Different parts of Tinospora cordifolia

CASSIATORA

In ancient system of medicine, C. tora was used to treat a variety of medical complications like bronchitis, constipation, conjunctivitis, ulcer, hypertension, hypercholestrolemic, liver damage. (19-21)

Material and Methods

ANIMALTRIAL

Collection of plant material

Anthocephalus indicus, Hibiscus rosa sinensis,

Tinospora cordifolia and Cassia tora were collected from the local area of Lucknow and identied taxonomically by the Department of Pharmacology, Era's Lucknow Medical College & Hospital, Lucknow and a voucher specimen were also submitted. (21)

Cassia tora

 

Plate 3.

 

 

 

V.S. of ower

3cm

ower

plant

4mm

fruit

seed

Preparation of extracts

Anthocephalus indicus (root & fruits), Hibiscus rosa

sinensis (root), Tinospora cordifolia (stem) and Cassia tora (seeds) were dried under shade and made into ne powder using laboratory mill. Powder (1000g) was extracted thrice with 3x2000 ml portions of 95% ethyl alcohol in a laboratory percolator at room temperature. Time allowed for each extraction was 8 hr. The extract obtained after third extraction was colorless. All the extracts were mixed; alcohol was distilled out at reduced temperature (20 0C) and reduced pressure (100 psi) in a rotor evaporator. This yielded 15g, 20g, 18g, and 20g (2% w/w) of crude extract, which was used for in vivo study. (21)

Preparation of doses

A quantity of 50 mg each (Anthocephalus indicus

(root), Hibiscus rosa sinensis (root), Tinospora cordifolia (stem) and Cassia tora (seeds)) extract were suspended /ml tripled distilled water (TDW) containing 2 % (w/v) gum acacia. The suspension were given in a volume of 1ml/100g animal bw (500 mg drug /kg bw) by oral intubation. (21)

Chemicals

Blood glucose (BLG), Total cholesterol (TC), triglycerides (TG), Phospholipid (PL) were analyzed using standard kits from Erba Diagnostic, (Mannheim GmbH, Germany) by an auto analyzer (Erba Mannheim, EM 360, Germany). Intralipid from victrum AB, In the Kabivitrum Group, Stockholm, Sweden. (21)

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VOL.4 NO.2

Experimental animals

Healthy male adult rats of Charles Foster strain (200- 250g) bred in the animal house of the Central Drug Research Institute, Lucknow were used. The animals were kept in controlled conditions; temperature 25 - 26˚C, relative humidity 60-70% and 12/12 hrs light / dark cycle (light from 08:00 AM to 08:00PM), provide with standard pellet diet (Lipton India Ltd.), and water adlibitum. (21)

Experimental design

The rats were divided in ve groups having six animals in each as follows: Group 1: normal control rats (on normal saline); Group 2, Anthocephalus indicus treated normal rats rats; Group 3, Hibiscus rosa sinensis treated normal rats; Group 4, Cassia tora treated normal rats; Group 5 Tinospora cordifolia treated normal rats. (21)

Assessment of biochemical parameters

The blood was centrifuged and plasma was separated. The fasting blood sugar (FBS) was analyzed in plasma, Super oxide dismutase (SOD), Catalase (CAT), Hepatic triglyceride lipase (TGL) and lipoprotein lipase (LPL) were estimated in liver homogenate. Serum total cholesterol (TC), triglyceride (TG), high density lipoprotein total cholesterol (HDL-TC), serum bilirubin, SGPT, SGOT and alkaline phosphatase were assayed by standard spectrophotometric methods. Low

density lipoprotein total cholesterol (LDL-TC) andvery low density lipoprotein total cholesterol (VLDL-TC) were calculated by Friedewald's equation. Serum was also used for the assay of lecithin cholesterol acyl transferase activity (LCAT), lipid peroxide (LPO), and reduced glutathione (GSH). A portion of serum was fractionated into very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) by polyanionic precipitation methods. Lipoproteins were measured for their total cholesterol (TC), phospholipids (PL), triglyceride (TG) and apoprotein by standard spectrophotometric methods. were analysed in liver homogenate. (22-38)

Statistical analysis: One-way-analysis of variance (ANOVA- Newman's Student t-test) was performed by comparison of values for normal treated group with normal healthy control. All hypothesis testing were two-tailed. P <0.05 was considered statistically signicant and the results were expressed as mean ± SD. The Graph pad INSTAT 3.0 software was used to carried out the statistical analysis. (39)

RESULTS

Effect of drug treatment on body weight of drug treated normal rats after 1 month with respect to Normal Control without drug treated

Weight of all experimental rats (Normal Control

Group

Body Weight (gm)

 

Percent change with

'p' value

 

 

 

respect to normal control

 

 

 

 

 

 

Normal

Study Day 0

242.50±33.90

-

-

Control+ 2

(Beginning)

 

 

 

% Aqueous

Study Day 30

360±23.56

-

-

gum acasia

(End of study)

 

 

 

 

 

 

 

 

Normal

Study Day 0

251.16±18.32

+3.71

NS

Control + A.

Study Day 30

345±20.70

-4.16

NS

indicus treated

 

 

 

 

 

 

 

 

 

Normal

Study Day 0

246.50±28.44

+1.65

NS

Control +

Study Day 30

320±27.66

-11.00

NS

Hibiscus treated

 

 

 

 

Normal

Study Day 0

247.16±27.31

+2.00

NS

Control +

Study Day 30

351±126.44

-2.50

NS

Cassia tora treated

 

 

 

 

Normal

Study Day 0

240.00±21.72

-.82

NS

Control + T.

Study Day 30

348±23.65

-3.00

NS

cordifolia treated

 

 

 

 

Table 1: Effect of drug treatment on body weight of normal rats after 1 month

 

Values expressed as gram (gm) are mean ± SD of six rats, drug treated groups were compared with Normal Control.

Percent change is with respect to Normal Control .NS = Not signicant.

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EFFECTS OF NATURAL PRODUCTS ON BODY WEIGHT AND BIOCHEMICAL PARAMETERS IN HEALTHY RATS

without drug treated and with drug treated) was record in beginning of study and after completion of the study. Percent change in weight was calculated with respect to healthy control in end of this study. No any signicant change in weight of drug treated normal rats was found with respect to normal control without drug treated, after one month, in end of the study (Table 1).

EFFECT OF A. INDICUS, H. ROSA SINENSIS, C. TORA AND T. CORDIFOLIA ON BLOOD GLUCOSE, SERUM LIPID AND LIPOPROTEIN PROFILE IN HEALTHY CONTROL RATS (NORMALRATS).

In normal rats, administration of A. indicus, H. rosa sinensis, C. tora and T. cordifolia at the dose of 500 mg/kg b.w. orally once daily for 30 days lowered the levels of blood glucose (0.5 – 4.0%), TC (2-4%), PL (2- 4%), TG (0.3-2.0%) and increase in total serum protein (1-3%) Table-2. Furthermore, treatment with test natural products the lipid and protein components of

serum lipoproteins remains almost the same to that of Control (Table-2).

It was seen that A. indicus. H. rosa sinensis, C. tora and T. cordifolia caused decrease in the level of - Lipoprotein TC (2-8%), PL (2-8%), TG (0.82-6.0%) and apolipoproteins (0.3-1.3%) respectively. However, herbal preparations increase lipid and protein contents of -Lipoprotein HDL by (0.25-6%) (Table-3).

EFFECTS OF A. INDICUS, H. ROSA SINENSIS, C. TORA AND T. CORDIFOLIA ON SERUM FREE FATTY ACIDS, HEPATIC TRIGLYCERIDE LIPASE AND TOTAL HEPATIC LIPOPROTEIN LIPASEACTIVITY STATUS IN NORMALRATS.

In normal rats administration of A. indicus, H. sinensis, C. tora and T. cordifolia for 30 days lowered the levels of free fatty acid (0.7-15%) hepatic triglyceride lipase (0.47-4%) and total hepatic lipoprotein lipase activity

(0.25-4%) respectively (Table 4).

Experimental

Blood

TC

PL

TG

Protein

schedule

glucose

(mg/dl)

(mg/dl)

(mg/dl)

(g/dl)

 

(mg/dl)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Normal Control

89.79±10.30

87.50±11.16

75.96±7.36

82.21±5.80

7.00±0.55

 

 

 

 

 

 

A. indicustreated

88.20±1060

85.65±11.69

74.62±7.53

81.61±5.76

7.09±0.56

 

(-2%)

 

(-2%)

(-2%)

(-1%)

(+1%)

H. rosa sinensis

86.64

±

10.09

85.88

±

11.55

74.95

±

81.35

±

7.11

±

0.58

treated

 

 

8.09

5.86

 

(-4%)

 

(-2%)

(-1%)

(-1%)

(+2%)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

C. tora treated

89.27

±

10.8

84.45

±

11.46

74.77

±

80.29

±

7.10

±

0.58

 

 

 

7.34

5.49

 

 

(-0.57%)

(-4%)

(-2%)

(-2%)

(-1%)

 

 

 

 

 

 

 

 

 

 

 

 

T. cordifolia

88.76 NS

84.67±11.60

74.89±7.46

81.89±6.04

7.18±0.53

treated

±10.15 (-1%)

(-3%)

(-2%)

(-0.38%)

(+3%)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Table 2: ffects of A. indicus, H. rosa sinensis, C. tora and T. cordifolia on blood glucose and serum lipids in normal rats

Values expressed are mean ± SD of six rats. Values with parenthesis are percentage change. Drug treated groups are compared with control (change are non signicant)

Experime

Very low density lipoprotein

Low density lipoprotein (LDL)

High density lipoprotein (HDL)

ntal

 

(VLDL)

 

 

 

 

 

 

 

 

 

schedule

TC

PL

TG

Apo-

TC

PL

TG

Apo-

TC

PL

TG

Apo-

 

 

 

 

protein

 

 

 

protein

 

 

 

protein

 

 

 

 

 

 

 

 

 

 

 

 

 

Normal

8.11

16.80

40.10

7.02

17.87

11.97

20.60

16.87

51.64

41.18

14.90

173.82

Control

±0.66 ±2.06 ±3.29

±0.99

±1.73 ±1.43 ±1.76

±2.37

±5.59 ±3.36 ±1.51

±11.63

 

 

 

 

 

 

 

 

 

 

 

 

 

A. indicus

7.71

16.06

39.77

7.01

17.40

11.07

19.90

16.92

54.78

41.95

15.07

174.08

treated

±0.54

±2.12

±3.43

±0.73

±1.66

±1.25

±1.42

±2.42

±8.27

±3.32

±1.49

±11.59

(-5%) (-4%) (0.82%) (-0.14%) (-3%) (-8%) (-3%) (+0.6%) (+6%) (+2%) (+1%) (+0.14%)

Table 3:Effect of A. indicus, H. rosa sinensis, C. tora and T. cordifolia on lipoprotein prole in normal rats

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July - Dec 2017ERA’S JOURNAL OF MEDICAL RESEARCHVOL.4 NO.2

H. rosa

7.72

15.93

39.72

6.94

17.51

11.55

20.02

16.82

51.91

41.72

15.08

174.21

sinensis

±

0.50

±

2.18

±

±

1.05

±

1.79

±

1.39

±

1.67

±

2.43

±

5.67

±

3.30

±

1.47

±

11.85

treated

 

 

3.39

 

 

 

 

 

 

 

 

 

(-5%)

(-5%)

(0.82%)

(-1.13%)

(-2%)

(-4%)

(-3%)

(-0.3%)

(+0.25%)

(+1.0%)

(+1%)

(+0.22%)

 

 

 

 

 

 

 

 

 

 

 

 

 

C. tora

7.58

16.46

39.37

7.02

17.19

11.46

19.30

17.08

53.50

42.25

14.89

172.17

treated

±0.53

±2.09

±3.42

±0.87

±1.71

±1.57

±27.03

±2.25

±8.78

±3.29

±1.10

±14.65

 

(-7%)

(-2%)

(1.82%)

(-0%)

(-4%)

(-4.25%)

(-6%)

(-1.2%)

(+4.0%)

(+3%)

(+0%)

(-0.94%)

 

 

 

 

 

 

No

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

change

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

T.

7.44

16.43

39.74

7.00

17.57

10.98

19.94

16.95

53.94

42.21

15.06

172.30

cordifolia

±0.52

±1.89

±3.35

±0.89

±2.17

±1.21

±1.69

±2.35

±4.36

±3.24

±0.94

±14.71

treated

(-8%)

(-2%)

(-0.82%)

(+0.85%)

(-2%)

(-8%)

(-3%)

(-0.5%)

(+4%)

(+2%)

(+1%)

(0.9%)

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Cont.....Table 3: Effect of A. indicus, H. rosa sinensis, C. tora and T. cordifolia on lipoprotein prole in normal rats

Values expressed are mean SD of six rats. Values with parenthesis are percentage change.Drug treated groups are

compared with control (changes were no signicant)

Experimental Schedule

Serum Free Fatty Acida

Hepatic Liprotein

Total Hepatic Lipoprotein

 

 

LipaseActivityb

Lipase Activityb

Normal Control

1.67±0.18

71.78±7.58

85.01±0.06

A. indicus treated

1.65±0.12(-1.0%)

74.65±7.48 (+4%)

88.53±4.58 (+4.0%)

 

 

 

 

H. rosa sinensis treated

1.61±0.18(4.0%)

74.20±7.83 (+3%)

85.30±8.98(+0.33%)

 

 

 

 

C. tora treated

1.62±0.19(-3.0%)

72.12±7.55 (+0.47%)

85.23±9.08(+0.25%)

 

 

 

 

T. cordifolia treated

1.60±0.19(-4.0%)

74.87±7.96(+4%)

88.70±5.12(+4%)

 

 

 

 

Table 4: Effect of A. indicus, H. rosa sinensis, C. tora and T. cordifolia on serum free fatty acid total hepatic

Lipoprotein lipase and hepatic Trigleceride lipase activity in normal rats.

Values are expressed as mean SD of six rats Drug treated groups are compared with control. (The changes are non

signicant); a=molFFA/L, b=molFFA released hr/mg protein

Experimental schedule

Serum Lipid peroxidea

Hepatic SODb

Hepatic CATb

Normal Control

2.91±0.42

2.78±0.19

3853±251.36

A. indicus treated

2.67±0.39

2.88±0.18

3910±267.08

 

(8.0%)

(+4%)

(+1.47%)

H. rosa sinensis treated

2.48±0.42

2.91±0.23

3916±302.23

 

(-15%)

(+5%)

(+1.5%)

C. tora treated

2.89±0.43

2.82±0.16

3929±331.42

 

(-0.7%)

(+1.36%)

(+2%)

T. cordifolia treated

2.46±0.39

2.89±0.17

3944±146.40

 

(-15%)

(+4%)

(+2.36%)

 

 

 

 

Table 5: Effect of A. indicus, H. rosa sinensis, C. tora and T. cordifolia on serum MDA and hepatic SOD, hepatic

Catalase activity in normal rats

Values are expressed as mean SD of six rats. Drug treated groups are compared with control. (The changes are non

signicant); a=molMDA/ml, b=units/min /mg protein

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EFFECTS OF NATURAL PRODUCTS ON BODY WEIGHT AND BIOCHEMICAL PARAMETERS IN HEALTHY RATS

 

Serum Bilirubina

SGPTb

SGOTb

ALPc

Normal control

0.62±0.06

22.29±2.74

57.22±7.35

17.63±0.9

A. indicus treated

0.50±0.10

20.73±2.91

56.65±6.90

17.34±1.09

 

(-4.83)

(-6.99)

(-0.99)

(-1.64)

Hibiscus treated

0.59±0.09

21.28±2.89

56.71±6.17

17.43±1.14

 

(-4.83)

(-4.53)

(-0.89)

(-1.13)

C. tora treated

0.570.08

20.71±2.91

56.30±6.92

17.26±1.07

 

(-8.06)

(-7.08)

(-1.60)

(-2.09)

T. cordifolia treated

0.58±0.13

21.87±2.80

56.03±6.86

17.37±1.08

 

(-6.45)

(-1.88)

(-2.07)

(-1.47)

Table 6: Effect of natural products on the hepato specic markers in the serum of normal rats.

Values are expressed as mean+SD of six rats, values in the parenthesis are percent change (changes are non signicant). Units a=mg/dl, b=units/l, c=units/dl.

EFFECT OF A. INDICUS, H. ROSA SINENSIS, C.

TORA AND T. CORDIFOLIA ON SERUM LIPID

PEROXIDE, HEPATIC SOD AND HEPATIC

CATALASE ACTIVITY IN NORMAL RATS

In normal rats administration of A. indicus, Hibiscus rosa sinensis C. tora and T. cordifolia for 30 days lowered the level of serum lipid peroxide (0.70-15.0%), and increase in the levels of hepatic SOD (1-5%), catalase (1-2.3%) respectively (Table 5).

EFFECT OF A. INDICUS, H. ROSA SINENSIS, C. TORA AND T. CORDIFOLIA ON HEPATO- SPECIFIC MARKER IN NORMAL CONTROL RATS

In normal rats administration of above mentioned herbal preparations for 30 days lowered the levels of Bilirubin, SGPT, SGOT, ALP in serum by (4-8%), (1- 6%), (0.6-2.07%) and (1-2%) respectively (Table 6).

Discussion:

In normal rats treatment with natural products (A. indicus, H. Rosa sinensis, C. tora, T. cordifolia) at the doses of 500 mg/kg body weight orally once in a day for 30 days did not alter signicantly, their body weight (Table 1) or blood biochemical parameters namely the levels of glucose, serum protein lipid prole (Table 2), lipoprotein prole (Table 3), free fatty acid with lipolytic enzyme activities (Table 4), lipid peroxide and antioxidant enzymes (Table 5). Also there was no signicant change in the hepatospecic parameters like Serum Bilirubin (S. Bil), Serum Glutamate Pyruvate Transminase (SGPT), Serum Glutamate Oxaloacetate Transminase (SGOT) and Alkaline Phosphatase (ALP) (Table 6). This indicated that the herbal preparations as such did not exert any adverse and toxic side effect during treatment in rats. The results of the present study demonstrated that natural products did not cause signicant decrease in biochemical parameters in

healthy normal rats. That's why it very clear here if healthy person will take natural products it never cause hypoglycemia, hypolipidemia and under weight. Natural products also not cause any signicant change on hepato-specic parameters. Thus from this study we conclude that, natural products are safe, non toxic and free from side effects, in comparison to synthetic drugs. (40-42)

Conclusion

It should be pointed out here that plant derived natural compounds have established a proven platform for developing new drug synthesis with fewer side effects or free from side effects.

Conict of interest statement

The authors declare that they have no conict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors. The study was approved by the Institutional Animal Ethics Committee of Central Drug Research Institute and was carried out in accordance with the current guidelines set by Organization for Economic Co-operation and Development (OECD), received from Committee for

the Purpose of Control and Supervision of

Experiments on Animals (CPCSEA), Ministry of Social Justice and Empowerment, Government of India for the care of laboratory animals

Acknowledgement: One of us (Vishnu Kumar) is grateful to the Director, Central Drug Research Institute (CDRI), Lucknow for experimental support, Director Academics, Era's Lucknow Medical College and Hospital, Lucknow for nancial support and Late Dr Ramesh Chander retired Scientist, Biochemistry, Division, CDRI, Lucknow for his expert guidance for the period of this research.

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How to cite this article : Kumar V., Mehdi F., Saxena J. K ., Singh R. K., Srivastava M.N., Ahmad S., Effects Of Natural Products On Body

Weight And Biochemical Parameters In Healthy Rats. EJMR2017;4(2):30-37.

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