Printer Friendly

Effect of Flax Seeds (Linum usitatissimum) on Uterine and Ovarian Protein Contents, Ovarian Cholesterol, Serum Estradiol and Onset of Puberty in Immature Female Mice.

Byline: SYED M. RAIHAN DILSHAD, NAJIB-UR-REHMAN, NAZIR AHMAD, ARSHAD IQBAL, MUHAMMAD AMJAD ALI AND ASIF AHMAD

ABSTRACT

The effects of aqueous methanolic extract of Flax seeds (FS) on body and organ weights, uterine and ovarian protein contents, ovarian cholesterol concentration, serum estradiol concentration, age at vaginal opening and estrus in immature female mice were investigated. One hundred and twenty immature female Balb/c mice (27 days old) were divided in to four equal groups A, B, C and D. Mice of group A served as untreated control, while mice of groups B, C and D were given orally the extract of FS at the dose rates of 100, 200 and 300 mg/kg body weight, respectively for 25 days. Onset of vaginal opening and estrus were monitored for mice of each group. Six mice of each group were euthanized at day 5, 10, 15, 20 and 25 of treatments. Overall mean values for body, ovarian and uterine weights were higher in mice given higher doses of extract (200 or 300 mg/kg) compared to control group or those given low dose (100 mg/kg; P less than 0.05).

The same was true for the ovarian and uterine protein contents, while an opposite trend was seen for ovarian cholesterol contents. In mice treated with FS, highest serum estradiol levels were reached 5-10 days earlier compared to mice of control group. Similarly, vaginal opening and estrus were recorded at an early age in mice given higher doses of extract. It was concluded that FS extract increased serum estradiol, uterine and ovarian protein contents, decreased ovarian cholesterol and enhanced onset of puberty in immature female mice. (c)2012 Friends Science Publishers

Key Words: Flax seeds; Mice; Organs weight; Ovarian and uterine proteins; Ovarian cholesterol; Serum estradiol; Puberty

INTRODUCTION

Livestock play an important role in agricultural economy of Pakistan. Late attainment of puberty is, however, one of the important factors adversely affecting the reproductive efficiency of livestock. Although modern facilities have replaced traditional practices in some parts of developed world, ethnoveterinary practices (EVP) still have significant contributions to animal health in areas where farmers either do not have access to the modern medicine or they have faith in the traditional medicine. In this regard, livestock farmers use various indigenous plants and other materials such as FS, dried dates, raw eggs etc. to hasten onset of puberty and improve fertility of dairy animals (Njidda and Isidahomen, 2011; Bilal et al., 2012).

Ethnopharmacological studies have shown that a number of medicinal plants produce secondary metabolites which may affect reproduction in man and animals (Sakamoto et al., 1988). Flax seeds/linseeds (Linum usitatissimum L.), locally called Alsi, are traditionally used as a protein supplement in livestock feed. Flax seeds exhibit antioxidant property (Rhee and Brunt, 2011), and are useful in preventing hyperlipidaemia and diabetic complications in rats (Makni et al., 2011). Flax seeds also suppress atherosclerosis (Prasad, 2009), and cardioprotective effects of lignan concentrate extracted from flax seeds have been seen in rats (Zanwar et al., 2011). Moreover, FS have been shown to reduce growth of human breast tumor cells (MCF-7) in anthymic mice (Chen et al., 2009).

Feeding Flax seeds (Linum usitatissimum) to dairy cows increases first service conception rate by 17% (Petit et al., 2001). A recent survey has shown that FS are used by the local farmers for the treatment of retention of foetal membranes, silent estrus and delayed puberty in cattle and buffaloes (Dilshad, 2009). The present study evaluates the effects of aqueous methanolic extract of FS on body weight, uterine and ovarian weights, uterine and ovarian protein contents, ovarian cholesterol concentration, serum estradiol concentration, and onset of puberty in immature female mice.

MATERIALS AND METHODS

Collection and extraction of flax seeds: Flax seeds were purchased from the local market and ground. The powder was used for the preparation of aqueous methanolic extract, using 70% aqueous methanol solution, as described elsewhere (Ahmad et al., 2012).

Experimental mice: One hundred and twenty (120) immature female Balb/c mice (20 days old) were obtained from the breeding colony being maintained at the National Institute of Health (NIH), Islamabad, Pakistan. At NIH, all pups were sorted by sex at birth (day 1) and redistributed so that all litters were standardized to 10 female pups per dam. Mice were weaned at day 19 of age. All animals were housed under controlled lighting (12 h light: 12 h dark) and temperature (22+-2degC) conditions. Mice were given fresh water and NIH-31 rodent diet (Table I) ad libitum. All animal procedures complied with NIH animal care guidelines.

After 7 days of acclimatization (27 days of age), mice were treated either with desired concentration of the extract (treatment groups) or distilled water (control group). The aqueous methanol extract was dissolved in distilled water to obtain desired concentration to be given to each group of experimental mice.

Experimental design: The experimental mice were divided into four groups A, B, C and D, with 30 mice in each group. Mice of group A served as control and were given distilled water. Mice of groups B, C and D were orally given the extract of FS at the dose rates of 100, 200 and 300 mg/kg body weight, respectively, for 25 days.

Post treatment monitoring: From the day of the start of treatment (day 27 of age), live weight for each mouse was recorded at 5 days intervals. Vaginal appearance of all mice was observed daily for coloration and degree of distention at the time of medication. After vaginal introitus, vaginal lavage was taken daily from each mouse to determine the age at first estrus (puberty) judged from the cornified smear (Cooper et al., 1993).

Blood collection: Six mice from each group were euthanized at day 5, 10, 15, 20 and 25 of treatments by cervical dislocation. For this purpose, mice were anesthetized in glass desiccators containing a swab of methoxyflurane (Metofane; Pitman-Moore, Inc.). Before killing, the blood was collected in a tube; the serum was separated and stored at 4degC for estradiol analysis.

Collection of uterus and ovaries: After incision of the skin and the abdominal muscles, uterus along with ovaries was removed. Both uteri and ovaries were blotted free of blood and weighed separately. Two ovaries from control and each treated group were randomly selected and fixed in Bouin solution for histological studies.

Estradiol quantification: Serum estradiol was quantified by using commercially available ELISA kit (Estradiol Enzyme Immuno Assay test kit, Catalog No: BC- 1111, BioCheck, Inc., USA), which had 100% cross reactivity for estradiol (E2). The BioCheck E2 EIA is based on the principle of competitive binding between E2 in the sample and E2-HRP conjugate for a constant amount of rabbit anti- Estradiol. After processing the samples and standards as per instructions of the manufacturer of kit, optical density of the samples and standards was determined at 450+-10 nm wavelength, using a Microsrtrip Reader (Stat-Fax-303, Awareness Technology, Inc.). The concentration of the hormone in the sample was determined from a standard curve generated between the concentrations and optical density of the standards.

Biochemical studies: The uterine and ovarian tissues were processed for the determination of proteins and cholesterol contents. For this purpose, ovaries and uterine samples were separately homogenized in 10 volumes of TS buffer (0.25 M sucrose, 1 mM EDTA and 10 mM Tris-HCl, pH 7.4). The homogenate was then centrifuged at 6000 x g at 4oC for 15 minutes and supernatant was used for the determination of proteins in both uteri and ovarian tissues and cholesterol contents in ovarian tissue (Telefo et al., 1998).

Protein contents in the ovarian and uterine tissues were estimated by using protein assay kit (Catalog No., 500- 0207; Quick Start Bovine Serum Albumin Standard set, 2x2 mL each, with concentrations of 2.0, 1.5, 1.00, 0.75, 0.5, 0.25 and 0.125 mg/mL). The microassay protocol for the protein determination was performed by using 300 uL microplate assays. Total protein level was expressed as mg/100 g fresh tissue weight.

Ovarian cholesterol concentrations were estimated by using Cholesterol assay kit (Breur and Breur Diagnostic Laboratories, USA) and expressed as mg/100 g fresh tissue weight. The principle of this assay kit is the determination of cholesterol after enzymatic hydrolysis and oxidation (Artiss and Zak, 1997) at 546 nm.

Histological studies: The preserved ovaries from mice of each group were dehydrated in graded alcohols, cleared in benzene, embedded in paraffin, serially sectioned at 5 um and then stained with hematoxylin and eosin. Ovarian sections were examined under light microscope for determining various developmental stages of follicles (Chen et al., 1981).

Statistical analysis: The data were analyzed by using General Linear Model (GLM) and means were compared through Least Significant Difference test. For this purpose, Statistical Package for Social Sciences (SPSS, Version 10, Chicago, IL, USA) was applied.

RESULTS AND DISCUSSION

Body weight: Oral administration of different doses of aqueous methanol extract of FS extract significantly affected the body weight (Table II). At the 5th day of treatment, mice of groups C and D showed higher body weight compared to those of groups A and B. This trend was maintained till the end of the study. On overall basis, the mice of group D were heaviest, followed by those of

Table I: Composition of mice feed (%)

Ingredients###Quantity

Corn starch###62.5

Corn oil###10.0

Glucose###5.0

Casein###12.5

Mineral +

vitamin mix###10.0

Total###100

Table II: Effects of different doses of aqueous methanolic extract of Flax seeds on body and organ weights of immature female mice (Mean +- SE)

Parameter###Treatment groups###Duration of treatment (Days)###Overall mean

###5###10###15###20###25###

Body weight (g)###A (Control)###18.69 +- 0.04###19.93 +- 0.02###22.44 +- 0.05###24.72 +- 0.02###24.89 +- 0.21###19.66 +- .08c

###B (100 mg/kg)###17.77 +- 0.43###19.76 +- 0.24###21.93 +- 0.15###24.42 +- 0.20###24.68 +- 0.50###19.36 +- 0.23d

###C (200 mg/kg)###20.28 +- 0.19###22.48 +- 0.15###25.85 +- 0.19###26.8 +- 0.21###27.50 +- 0.20###21.43 +- 0.16b

###D (300 mg/kg)###20.43 +- 0.19###23.79 +- 0.12###26.03 +- 0.11###26.82 +- 0.11###28.57 +- 0.12###21.81 +- 0.12a

###Mean###19.29 +- 0.21E###21.49 +- 0.13D###24.06 +- 0.13C 25.69 +- 0.18B###26.41 +- 0.26A###

Ovarian weight (mg)###A (Control)###7.43 +- 0.07###11.27 +- 0.09###12.70 +- 0.12###14.43 +- 0.12###17.30 +- 0.07###12.63 +- 0.09d

###B (100 mg/kg)###7.57 +- 0.09###11.64 +- 0.12###13.39 +- 0.11###17.75 +- 0.11###17.82 +- 0.11###13.63 +- 0.11c

###C (200 mg/kg)###12.42 +- 0.12###18.65 +- 0.12###18.87 +- 0.09###19.22 +- 0.13###18.88 +- 0.16###17.61 +- 0.12b

###D (300 mg/kg)###12.53 +- 0.10###19.29 +- 0.11###19.28 +- 0.04###19.44 +- 0.09###19.56 +- 0.11###18.02 +- 0.09a

###Mean###9.99 +- 0.095E###15.21 +- 0.11D###16.06 +- 0.09C 17.71 +- 0.11B###18.39 +- 0.11A###

Uterine weight (mg)###A (Control)###24.3 +- 0.09###77.55 +- 0.148###69.82 +- 0.14 64.94 +- 0.102###62.77 +- 0.199###59.88 +- 0.135d

###B (100 mg/kg)###25.73 +- 0.11###79.39 +- 0.145###72.57 +- 0.217###76.77 +- 0.14###71.85 +- 0.54###65.26 +- 0.23c

###C (200 mg/kg)###74.8 +- 0.12###77.41 +- 0.07###82.23 +- 0.103###84.22 +- 0.13###93.68 +- 0.126###82.47 +- 0.11b

###D (300 mg/kg)###80.49 +- 0.16###78.71 +- 0.181###92.97 +- 0.198###96.7 +- 0.179 106.07 +- 0.579###90.99 +- 0.26a

###Mean###51.33 +- 0.12E 78.27 +- 0.136D###79.4 +- 0.165C 80.66 +- 0.138B 83.59 +- 0.361A

Table III: Effects of different doses of aqueous methanolic extract of Flax seeds on tissue biochemical metabolites and serum estradiol in immature female mice (Mean +- SE)

Parameter###Treatment groups###Duration of treatment (Days)###Overall mean

###5###10###15###20###25###

Uterine###protein###A (Control)###11.02 +- 0.02###14.33 +- 0.02###18.20 +- 0.01###18.61 +- 0.03###18.37 +- 0.04###16.11 +- 0.03d

contents (mg/100g)###B (100 mg/kg)###11.55 +- 0.02###15.07 +- 0.03###18.96 +- 0.03###19.00 +- 0.037###19.12 +- 0.02###16.74 +- 0.03c

###C (200 mg/kg)###14.43 +- 0.04###18.26 +- 0.02###19.18 +- 0.02###19.77 +- 0.02###20.22 +- 0.02###18.37 +- 0.02b

###D (300 mg/kg)###18.20 +- 0.03###18.33 +- 0.05###19.58 +- 0.04###19.86 +- 0.02###20.48 +- 0.02###19.29 +- 0.03a

###Mean###13.80 +- 0.03E###16.50 +- 0.03D###18.98 +- 0.03C###19.31 +- 0.03B###19.55 +- 0.02A###

Ovarian protein###A (Control)###8.32 +- 0.01###10.23 +- 0.01###11.92 +- 0.01###12.03 +- 0.05###12.08 +- 0.02###10.92 +- 0.02d

contents (mg/100g)###B (100 mg/kg)###8.48 +- 0.01###11.83 +- 0.02###11.94 +- 0.01###12.19 +- 0.03###12.74 +- 0.01###11.44 +- 0.02c

###C (200 mg/kg)###10.44 +- 0.02###12.00 +- 0.03###12.64 +- 0.01###12.98 +- 0.02###13.05 +- 0.02###12.22 +- 0.02b

###D (300 mg/kg)###12.02 +- 0.02###12.37 +- 0.02###13.01 +- 0.03###13.31 +- 0.01###13.46 +- 0.02###12.83 +- 0.02a

###Mean###9.82 +- 0.01E###11.61 +- 0.02D###12.38 +- 0.02C###12.63 +- 0.02B###12.83 +- 0.02A###

Ovarian cholesterol###A (Control)###3.54 +- 0.01###2.67 +- 0.01###2.13 +- 0.01###1.92 +- 0.01###1.79 +- 0.01###2.41 +- 0.01a

contents (mg/100g)###B (100 mg/kg)###3.38 +- 0.01###2.27 +- 0.01###2.03 +- 0.03###1.88 +- 0.01###1.74 +- 0.01###2.26 +- 0.01b

###C (200 mg/kg)###2.34 +- 0.01###1.88 +- 0.01###1.82 +- 0.01###1.77 +- 0.01###1.73 +- 0.01###1.91 +- 0.01c

###D (300 mg/kg)###1.91 +- 0.01###1.79 +- 0.01###1.74 +- 0.01###1.71 +- 0.01###1.73 +- 0.01###1.78 +- 0.01d

###Mean###2.79 +- 0.01A###2.15 +- 0.01B###1.93 +- 0.01C###1.82 +- 0.01D###1.75 +- 0.01E###

Serum estradiol###A (Control)###17.16 +- 0.08###13.33 +- 0.07###26.38 +- 0.07###35.60 +- 0.076###37.84 +- 0.09###26.06 +- 0.08b

contents (pg/ml)###B (100 mg/kg)###14.50 +- 0.08###20.70 +- 0.09###26.39 +- 0.09###37.62 +- 0.11###27.51 +- 0.07###25.34 +- 0.09c

###C (200 mg/kg)###12.35 +- 0.09###28.46 +- 0.09###38.45 +- 0.10###24.25 +- 0.02###26.23 +- 0.05###25.95 +- 0.07b

###D (300 mg/kg)###12.74 +- 0.14###26.87 +- 0.67###22.77 +- 1.33###37.18 +- 0.38###34.06 +- 0.36###26.73 +- 0.58a

###Mean###14.19 +- 0.10E###22.34 +- 0.23D###28.50 +- 0.40C###33.66 +- 0.15A###31.41 +- 0.14B

group C and A. However, the mice of B group attained lesser weight than the control and the mice of other two groups (P less than 0.05). This indicates that significant increase in the body weight was obtained with the administration of higher doses (200 and 300 mg/kg) of the FS extract. Similar observations have been recorded by Ahmad et al. (2012) in rats treated with aqueous methanolic extract of FS.

Ovarian weight: Maximum increase in ovarian weight was recorded between day 5 and 10 of treatment in mice of D and C groups, while maximum ovarian weight gain in group B was between day 15 and 20 of treatment. However, mice of group A attained maximum ovarian weight during the day 20 and 25 (Table II). Thus, higher doses of extract accelerated ovarian growth at an early age compared to low

As far as could be ascertained, this is the first report on the effect of FS on uterine or ovarian protein contents. Ahmad et al. (2012), however, observed a significant increase (P less than 0.05) in the serum total protein contents in immature rats given FS extract compared to controls. Whether this increase in the total protein contents in the blood is associated with increased total protein levels in the uterus or ovaries is not clear.

Ovarian cholesterol content: With increasing dose rate, cholesterol content in the mice ovaries decreased as the duration of treatment was increased. Overall minimum mean value of ovarian cholesterol content (mg/100 g ovary) was recorded in mice of group D (1.78 +- 0.01) and maximum was in control group A (2.41 +- 0.01), the difference being significant (P less than 0.05; Table III).

Thus, the ovarian cholesterol was low in the mice groups given FS extract. The possible reason may be that steroidogenesis is dependent on the availability of cholesterol substrate. Even if other factors such as gonadotropin stimulation and expression of steroid biosynthetic enzymes are in place, insufficient cholesterol substrate can limit steroidogenesis. Ovaries rely on a combination of substrate sources to support steroidogenesis, including cholesterol uptake from plasma lipoproteins, mobilization of intracellular cholesterol ester stores, and de novo cholesterol synthesis (Wade et al., 2002). The utilization of cholesterol in steroidogenesis decreased the amount of available cholesterol content in early pubertal mice. Chinoy and Patel (2001) also recorded that a block in the steroidogenic pathway in mice treated with fluoride or aluminium individually or in combination resulted in significant accumulation of cholesterol in the ovaries.

Previous studies (Guan et al., 2006; Abuelgassim, 2010) have also indicated that phytoestrogens of kudzu and soybean decreased total cholesterol and LDL-cholesterol, while HDL-cholesterol was not affected. Moreover, Flax seeds prevent hyperlipidaemia in diabetic rats (Makni et al.,2011).

Serum estradiol concentrations: Highest mean estradiol concentration (37.84+-0.09 pg/mL) in group A was observed at day 25 of treatment, while it occurred at day 20 in mice of group B (37.62 +- 0.11 pg/mL) and D (37.18 +- 0.37 pg/mL) and at day 15 in mice of group C (38.45 +- 0.10 pg/mL). It shows that in mice treated with FS, highest serum estradiol levels were reached 5-10 days earlier compared to mice of control group. On overall basis, serum estradiol was the highest in mice of group D and lowest in those of group B, differences among all groups were significant (Table III). The increased estradiol levels in mice treated with FS can be attributed to their phyto-estrogenic activity.

The current results support those of Tou et al. (1999) and Ahmad et al. (2012), who observed increased estradiol levels in rats treated with FS compared to controls. Similar activity of FS has also been reported by Richter et al. (2010) in stimulation of breast carcinoma cells (MCF7) with FS extract through estradiol production.

Vaginal opening and estrus: Effect of feeding 3 concentrations of FS extract to mice was variable for the onset of vaginal opening (VO) and estrus. Vaginal opening was seen at significantly lower age (P less than 0.05) in mice of groups D (30.5 days) and C (31.2 days) compared to mice of groups B (33.4 days) and A (34.9 days). The difference in the age of vaginal opening between groups C and D and A and B was non-significant. This indicates that vaginal opening occurred at lower age in mice given 200 or 300 mg/kg FS extract compared to mice of control group or those given low dose (100 mg/kg) of the extract. A similar trend was seen for the onset of first estrus (puberty), recorded at the lowest age of 34.1 days in mice of group D and the highest age of 48.8 days in the control group.

Onset of puberty in a female is the complex interaction between age, body weight, ovarian and uterine growth and secretion of gonadotropins including FSH and LH. During the pre-pubertal period, increased concentrations of FSH and LH stimulate growth and development of follicles on the ovaries. Previously, it has been reported that larger doses of estrogen suppress the secretion of pituitary gonadotropins and thus inhibit the follicular growth, whereas small doses of estrogens were found to enhance the follicular development (Smith and Bradbury, 1961). The increased ovarian weight and follicular development recorded in mice given FS extracts may be explained as the phytoestrogenic properties of the plant extracts which have been reported to possess weak estrogenic effects. Thus, mice treated with high doses of FS extract showed higher ovarian and uterine growth, showed higher serum estradiol levels earlier and attained puberty earlier compared to mice of control group.

Histological examination of the ovaries also revealed the presence of corpora lutea and growing follicles (Fig. 1) in the ovarian sections of mice of group D as early as day 5 and day 10 after treatment (32-37 days of age). In mice of group C, mature Graafian follicles (Fig. 2) were seen at Day 15 of treatment (42 days of age). Only developing follicles were seen in mice of groups B and A at day 42 of age (Fig. 3), while mature follicles in mice of these two groups were seen at day 20 of treatment (47 days of age). Murthy et al. (1997) found that immature mice showed early vaginal opening, premature cornification of vagina and increased uterine weight when treated with benzene extract of Hibiscus rosa sinensis flowers @ 125 and 250 mg/kg body weight intraperitoneally.

In conclusion, FS extract increases serum estradiol, uterine and ovarian protein contents, decreased ovarian cholesterol and enhanced onset of puberty in immature female mice.

REFERENCES

Abuelgassim, A.O., 2010. Effect of Flax seeds and date palm leaves extracts on serum concentrations of glucose and lipids in alloxan diabetic rats. Pakistan J. Biol. Sci., 13: 1141-1145

Ahmad, N., Z.U. Rahman, N. Akhtar and S. Ali, 2012. Effects of aqueous methanolic extract of Flax seeds (Linum usitatissimum) on serum estradiol, progesterone, kidney and liver functions and some serum biochemical metabolites in immature female rats. Pakistan Vet. J., 32: 211-215

Artiss, J.D. and B. Zak, 1997. Measurement of cholesterol concentration. In: Rifai, N., G.R. Warnik and M.H. Dominiczak (eds.), "Handbook of Lipoprotein Testing," pp: 99-114. AACC Press, Washington, USA

Bilal, T., F.E. Gursel, A. Ates and O. Keser, 2012. Effects of dietary b- glucan on serum lipids and performance indices in rats fed a diet enriched with cholesterol. Pakistan Vet. J., 32: 97-100

Chen, Y.T., D.R. Mattison, L. Feigenbaum, H. Fukui and J.D. Schulman, 1981. Reduction in oocyte number following prenatal exposure to a diet high in galactose. Science, 214: 1145-1147

Chen, J., J.K. Saggar, P. Corey and L.U. Thompson, 2009. Flax seed and pure seciosolariciresinol diglucoside, but not Flax seed hull, reduce human breast tumor growth (MCF-7) in anthymic mice. J. Nutr.,139: 2061-2066

Chinoy, N.J. and T.N. Patel, 2001. Effects of sodium fluoride and aluminium chloride on ovary and uterus of mice and their reversal by some antidotes. Fluoride, 34: 9-20

Circosta, C., R. Sanogo and F.M. Occhiuto, 2001. Effects of Calotropis procera on estrus cycle and oestrogenic functionality in rats. Pharmacology, 56: 373-378

Dilshad, S.M.R., 2009. Documentation of ethno-veterinary practices in Sargodha district (Pakistan) and investigations on the effects of some traditionally used plants on the puberty onset in mice. Ph.D. Thesis, University of Agriculture, Faisalabad, Pakistan

Cooper, R.L., J.M. Goldman and J.G. Vandenbergh, 1993. Monitoring of the estrous cycle in the laboratory rodent by vaginal lavage. In:

Heindel, J.J. and R.E. Chapin (eds.), Methods in Toxicology: Female Reproduction Toxicology, Vol. 3B, pp: 45-56. Academic Press, San Diego, USA

Guan, L.S., Y. Yeung, Y. Huang and Z.Y. Chen, 2006. Both soybean and kudzu phytoestrogens modify favourably the blood lipoprotein profile in ovariectomized and castrated hamsters. J. Agric. Food Chem., 54: 4907-4912

Hafez, E.S.E. and B. Hafez, 2006. Reproduction in Farm Animals, 7th edition. Lea and Febiger, Philadelphia, USA

Njidda, A.A. and C.E. Isidahomen, 2011. hematological parameters and carcass characteristics of weanling rabbits fed sesame seed meal (Sesamum indicum) in a semi-arid region. Pakistan Vet. J., 31: 35-39

Katzenellenbogen, B.S., H.S. Bhakoo, E.R. Ferguson, H.C. Lan, T. Tatee, T.L.S. Tsia and J.A. Katzenellenbogen, 1979. Estrogen and anti estrogen action in reproductive tissues and tumors. Recent Prog. Hormone Res., 35: 259-292

Makni, M., M. Sefi, El. M. Garoui, H. Fetoui, T. Boudawara and N. Zeghal, 2011. Dietary polyunsaturated fatty acid prevents hyperlipidemia and hepatic oxidant status in pregnant diabetic rats and their macrosomic offspring. J. Diabetes its Complications, 25: 267-274

Murthy, D.R., C.M. Reddy and S.B. Patil, 1997. Effect of benzene extract of Hibiscus rosa sinensis on the estrous cycle and ovarian activity in albino mice. Biol. Pharma. Bull., 20: 756-758

Ostrovsky, D., 1962. Estrogen-like substances in legumes and grasses. The influence of fractionation and route of administration on the estrogenic activity of plant materials. Canadian J. Biochem. Phys.,40: 159-164

Ostrovsky, D. and W.D. Kitts, 1963. The effect of estrogenic plant extracts on the uterus of the laboratory rat. Canadian J. Anim. Sci., 43: 106-112

Petit, H.V., R.J. Dewhurst, J.G. Proulx, M. Khalid, W. Haresign and H.Twagiramunugu, 2001. Milk production, milk composition and reproductive function of dairy cows fed different fats. Canadian J.Anim. Sci., 81: 263-271

Rhee, Y. and A. Brunt, 2011. FS supplementation improved insulin resistance in obese glucose intolerant people: a randomized crossover design. Nutr. J., 10: 44

Richter, D.U., S. Abarzua, M. Chrobka, C. Scholz, C. Kuhn, S. Schulze, M.S. Kupka, K. Friese, V. Briese, B. Piechulla, and U. Jeschke,2010. Effects of phytoestrogen extracts isolated from Flax on estradiol production and ER/PR expression in MCF7 breast cancer cells. Anticancer Res., 30: 1695-1699

Prasad, K., 2009. Flaxseed and cardiovascular health. J. Cardiovasc.Pharmacol., 54: 369-377

Sakamoto, S., H. Kudo, T. Kawasaki, R. Kuwa, N. Kasaharo, S. Sassa and R. Okamoto, 1988. Effects of Chinese herbal medicine, keshi- bukuryo-gan on the gonadal system of rats. J. Ethnopharmacol., 23:151-158

Smith, B.D. and J.T. Bradbury, 1961. Ovarian weight response to varying doses of estrogens in intact and hypophysectomized rats. Proc. Soc. Exp. Biol. Med., 107: 946-953

Telefo, P.B., P.F. Moundipa, A.N. Tchana, C.T. Dzickotze and F.T. Mbiapo, 1998. Effects of an aqueous extract of Aloe buettneri, Justicia insularis, Habiscus macranthus and Dicliptera verticilata on some physiological parameters of reproduction in immature female rats. J. Ethnopharmacol., 63: 193-200

Thompson, L.U., J. Chen, E. Hui, J. Mann and T. Ip, 2004. Interactive effects of Flax seeds and tamoxifen on human breast cancer. Proc. 60th Congress of Flax Institute, pp: 86-90. Fargo, ND, USA

Tou, J.C., J. Chen and L.U. Thompson, 1999. Dose, timing and duration of Flax seed exposure affect reproductive indices and sex hormone levels in rats. J. Toxicol. Environ. Heath A, 56: 555-570

Wade, R.L., R.A. Van Andel, S.G. Rice, C.L. Banka and C.A. Dyer, 2002.Hepatic lipase deficiency attenuates mouse ovarian progesterone production leading to decreased ovulation and reduced litter size. Biol. Reprod., 66: 1076-1082

Zanwar, A.A., M.V. Hedge and S.L. Bodhankar, 2011. Cardioprotective activity of flax lignan concentrate extracted from seeds of Linum usitatissmum in isoprenalin induced myocardial necrosis in rats. Interdiscip. Toxicol., 4: 90-97
COPYRIGHT 2012 Asianet-Pakistan
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2012 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:Dilshad, Syed M. Raihan; Najib-Ur-Rehman; Ahmad, Nazir; Iqbal, Arshad; Ali, Muhammad Amjad; Ahmad, A
Publication:International Journal of Agriculture and Biology
Article Type:Report
Geographic Code:9PAKI
Date:Oct 31, 2012
Words:4674
Previous Article:Germination and Growth Response of Rice and Weeds to Herbicides under Aerobic Conditions.
Next Article:Zinc-enriched Farm Yard Manure Improves Grain Yield and Grain Zinc Concentration in Rice Grown on a Saline-sodic Soil.
Topics:

Terms of use | Privacy policy | Copyright © 2022 Farlex, Inc. | Feedback | For webmasters |