Effect of Emex spinosa Campd extract on male fertility

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and some biochemical parameters in rats

Abstract: The effects of aqueous ethanolic extract of Emex spinosa on male fertility and some biochemical parameters (hepatoprotective activity and blood sugar) in rats were investigated. The results revealed that ethanolic extract of E. spinosa showed significant effects on serum ALT, AST, T. protein, T. bilirubin and blood glucose. Also, it showed significant effects on weight of reproductive organs, sperm count and testosterone.

Key words: Emex spinosa, hepatoprotective activity, male fertility.

Introduction:

Emex spinosa (L.) Campd.; an annual stout herb with spreading stems and sweet roots. Arabic name: Hambaz, Hambaizan, Rashaa, common in winter used as purgative and diuretic, the boiled leaf is used by African tribes for the cure of dyspepsia and biliousness, and to stimulate appetite (Watt 1962 and Mossa el al.1987). Emex spinosa is regarded as a minor weed, although it is well suited to the climate of southern Australia, and south-western Australia in particular, it is found also from Arabia to India (Holm et al., 1979). Emex spinosa is edible by the local inhabitants who plucked it and eat its petiole and carrot-like tap root (Mandaville 1990). Its phytochemical investigation revealed the presence of alkaloides, anthraquinones, coumarins and flavonoides (Rizk 1986). Five anthraquinone pigments were detected in this plant: chrysophenol, physcion, emodin, stigmasterol and campesterol in additional to β- sitosterol and B-D- glucoside (Abdel-Fattah et al. 1990). It is one of the important medicinal plants used to relief dyspepsia; stimulate appetite, and a remedy for stomach disorders and to relief colic. It is believed to be purgative and diuretic (Watt & Breyer-Brandwjik 1962).

Chloroform extract of Emex spinosa. Campd. displayed an elevated potency against Alternaria alternata. (Fries) Keissler (27.9 mm) and Saccharomyces cerevisiae. (27.5 mm). Zone of inhibition against other fungi varied from 19.9 mm to 25.9 mm, indicating a novel class of metabolite. Mandeel and Taha 2005.

A New α-methylanthraquinone glucoside, laccaic acid D 8- o -β-D-glucoside, and six known anthraquinones have been isolated from the aqueous alcoholic extract of Emex spinosus (L.) Campd. (Polygonaceae), together with two known flavonol glycosides (Hawas et al., 2006).

Aqueous and methanolic extracts of 18 leafy vegetables from South Africa were investigated for their, free radical scavenging activity, total phenolic content and anti-inflammatory properties. Out of 18 plants studied for anti-inflammatory activity, methanolic extracts from eight plants showed significant inhibition of 5- lipoxygenase (5-Lox) activity. Among the plants studied Bidens pilosa (IC50 21.8 μg/ml) showed maximum anti-inflammatory activity while Emex australis (IC50 81.4 μg/ml) showed minimum inhibition of 5-Lox activity. Percent scavenging activity of methanolic extract of Emex australis was 78.6%, Total phenolics (mg/g) in methanolic extract (27.4±0.36) (Akula and Odhav 2008).

The aqueous ethanolic extracts (70%) of Laurus nobilis and Emex spinosus leaves exhibited free radical scavenging action against 1,1-diphenyl-2-picrylhydrazyl (DPPH). Their concentrations of 50% inhibition (IC50) were 25.3 and 20.73 μg/mL, respectively, and isolated luteolin and rutin from this plant, these two compounds had strong scavenging activity with an IC50 of 4 and 4.6 μg/mL, respectively (Emam et al., 2010).

Material and Methods:

Plant Material. Emex spinosa was collected in the ---------The collected plant, identified by Prof. Dr. ------------, the plant was dried under shade and then grinded to fine powder.

Extraction.

The dried powder (aerial parts and roots) 1k g was extracted with 70% aqueous ethanol by percolation in the solvent with occasional shaking for 72 hour using three liters. The combined ethanolic extracts were concentrated under reduced pressure at a temperature not exceeding 35 oC.

Toxicological Studies.

LD50 values of the tested extracts were determined by a known method (Finney 1964)19. Using groups of rats 6 in each. Animals were kept under observation for 24 hr during which symptoms of toxicity and rate of mortality in each group were recorded from which the LD50.

Determination of Prolactin:

Prolactin was determined according to the method described by Tietz (1995).

Determination of Testosterone:

Testosterone was determined according to the method described by Chen et al., (1991)

Determination of (FSH).

Follicle-stimulation Hormone was determined according to the method described by Uotila et al., (1981)

Determination of Luteinizing Hormone (LH).

Luteinizing Hormone was determined according to the method described by Uotila et al., (1981).

Results and discussion:

Toxicological Studies

• Acute toxicity and median lethal dose (LD50) test:

Emex spinosa in oral doses up to 5000 mg/kg b.wt. did not produce any demonstrable acute toxic effects or deaths in all groups of mice.

Effect of prolonged administration:

Table 1: Effect of prolonged oral administration of ethanolic extract of Emex spinosa for 6 successive weeks on the serum activity of ALT and AST and serum levels of total bilirubin, total protein, albumin and glucose in rats, (n=6).

Groups

ALT

(UL-1)

AST

(UL-1)

T. bilirubin (mg dL-1)

T. protein (g dL-1)

Albumin (g dL-1)

Glucose

(mg dL-1)

Control

62.57±2.74

139.28±4.57

1.63±0.10

8.3±0.25

3.5±0.16

115.35±4.96

Emex spinosa

58.87±2.86

130.16±4.22

1.70±0.11

7.8±0.21

3.3±0.15

86.05±3.27***

Groups

Urea (mg dL-1)

Creatinine(mg dL-1)

Control

39.20±1.56

0.38±0.01

Emex spinosa

42.46±1.19

0.41±0.02

Table 2: Effect of prolonged oral administration of ethanolic extract of Emex spinosa for 6 successive weeks on the serum levels of urea and creatinine in rats, (n=6).

Hepatoprotective activity:

Table 3: Effect of the ethanolic extract of Emex spinosa on the serum activity of ALT and AST and serum levels of total bilirubin, total protein, albumin and glucose in rats with CCl4 induced - hepatotoxicity.

Groups

ALT

(UL-1)

AST

(UL-1)

T. bilirubin (mg dL-1)

T. protein (g dL-1)

Albumin

(g dL-1)

Glucose

(mg dL-1)

Control

372.5±8.34

447.6±11.64

3.6±0.18

5.1±0.21

2.5±0.18

221.4±7.12

Emex spinosa

234.5±7.15***

333.8±8.40***

2.9±0.16*

6.7±0.24***

3.2±0.22*

184.5±6.58**

Effect on male fertility:

Table 4: Effect of oral administration of ethanolic extract of Emex spinosa for 6 successive weeks on weights of sexual organs of male rats, (n=6).

Groups

Initial b.wt (g)

Final b.wt (g)

Weight of reproductive organs (g/100 g b.wt)

Testes (Pair)

Seminal vesicles

Ventral prostate

Control

230.2±7.42

246.8±7.42

1.67±0.06

0.62±0.02

0.47±0.02

Emex spinosa

226.7±7.50

235.4±7.59

1.94±0.08*

0.71±0.03*

0.54±0.02*

Table 5: Effect of oral administration of ethanolic extract of Emex spinosa for 6 successive weeks on semen characteristics of male rats, (n=6).

Groups

Sperm count (X 106/mL)

Sperm motility (%)

Unstained (live) sperms (%)

Total sperm abnormality (%)

Control

64.15±2.63

92.3±3.11

92.0±3.58

3.37±0.21

Emex spinosa

73.56±2.84*

93.5±3.27

93.3±3.25

2.75±0.18*

Table 6: Effect of oral administration of ethanolic extract of Emex spinosa for 6 successive weeks on plasma levels of reproductive hormones of male rats, (n=6).

Groups

Testosterone (ng mL-1)

Prolactin (ng mL-1)

FSH

(mIU mL-1)

LH

(mIU mL-1)

Control

4.11±0.38

0.65±0.02

7.47±0.22

0.63±0.02

Emex spinos

6.53±0.45**

0.63±0.03

7.17±0.28

0.60±0.04

Table 7: Effect of oral administration of ethanolic extract of Emex spinos on the percentages of pregnancies obtained with normally cycling rats, (n=10).

Groups

Pregnancies

n

%

Control

8

80

Emex spinosa

9

90

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