Evaluation of Achyranthes aspera root extract in treatment of infertility in male rats

Evaluation of Achyranthes aspera root extract in treatment of infertility in male rats

Abstract and Introduction:

To evaluate infertility in albino rats, researchers typically assess several key parameters including: sperm analysis (count, motility, morphology), hormonal levels (testosterone, FSH, LH), testicular weight and histology, accessory sex organ weights, and mating trials with female rats to determine pregnancy rates; all of these measurements are taken to identify potential disruptions in spermatogenesis, hormone balance, and overall reproductive function within the male albino rat model. 

Ayurveda is gaining popularity because it effectively treats many chronic illnesses. Ayurvedic remedies are often used in conjunction with and/or after traditional medical techniques as the majority of patients start taking conventional pharmaceuticals as soon as their diagnoses are made. Understanding food, spices, and medicinal plants' actions in-depth is necessary to fully appreciate their potential effect. The use of highly concentrated products derived from single plants, sometimes in the form of teas or pills, is of more concern, even though society uses ayurvedic herbs and Indian spices frequently and without incident. The methods by which polyherbal

medications and their extracts work differ from those of single ingredients or synthetic pharmaceuticals in several

important ways. Even though ayurvedic medications are made from natural herbal ingredients, their effectiveness

depends on how they are administered, taking into account the requirements of the patient and the nature of the ailment being treated. A member of the Amaranthaceae family, Achyranthes aspera Linn. Also known as Apamarga in Ayurveda is a crucial medicinal plant. The followinga rticle provides information about Apamarga, including its history, Ayurvedic properties, activities, and medical usefulness as stated in various sources.

KEYWORDS: Ayurveda, ayurvedic herbs, Apamarga, Ayurvedic properties, activities, infertility, testosterone

S.E.T'S COLLEGE OF PHARMACY, RAKESH MAHATMA SINGH, RGUHS, BENGALURU, KARNATAKA, INDIA

MATERIALS AND METHODS

Preparation of Achyranthes aspera extract

Fresh stems and roots of A. aspera were collected and dried under shade and then in an oven at a temperature <50°C. The stems and roots were ground into a fine powder. Then, 100 g of powdered stems and roots were boiled in 2000 ml of distilled water separately in two flasks for 2 h. One thousand and three hundred milliliters of filtrate was obtained from the stems and roots, which was then filtered using a filter paper. It was reduced to obtain a solid residue of 6 g by heating it at 60°. The solid residue obtained was then stored at a low temperature. Dimethyl sulphoxide was used to dissolve the solid residue made from the stems and roots to make different concentrations. One gram of solid residue was mixed in 10 ml of dimethyl sulphoxide to make 10% concentration and it was kept as a stock solution. From this stock solution, further different concentrations were made.

Animals

Adult male albino rats weighing 160 ± 10 g were used in the present study. The animals were kept in wire bottomed cages in a room under standard conditions of illumination with a 12-hour light–dark cycle at 23 ± 1°C. They were provided with tap water and a balanced diet ad libitum.

Analysis of sperm parameters :

Epididymal sperm were obtained by chopping cauda epididymis in 5.0 ml of Ham’s F12 medium. The sperm were counted using a Neubauer Chamber as describe by Belsey et al. Progressive sperm motility was evaluated by a previously described method within five minutes following their isolation from cauda epididymis at 37°C and the data were expressed as percent motility.

The morphological abnormalities in sperm were enumerated by the methodology reported by Hemavathi and

Rahiman  (using light microscopy)

Genetic analysis

DNA extraction

Isolation of genomic DNA from caudal epididymal spermatozoa was carried out using the protocol of Gebert et al.Briefly, sperm cells were lysed in 500 μl of a buffer consisting of 50 mM Tris–HCl at pH 8.0, 100 mM NaCl2,100 mM EDTA, 1% SDS and treated with 2.5 μl of Triton X-100 (Merck, Darmstadt, Germany), 21 μl of dithiothreitol (1 M) (Sigma-Aldrich Chemie) and 40 μl of proteinase K (10 mg/ml). DNA precipitation was performed in a saturated sodium chloride solution with subsequent addition of 100% ethanol (Roth, Hamburg, Germany).

The concentration of DNA and its relative purity were determined using a spectrophotometer based on absorbance at 260 and 280 nm, respectively. The integrity of extracted genomic DNA was verified by electrophoresis in 0.8% agarose gel using a DNA molecular weight marker (Eurblio, Paris, France).

Reference:

Belsey MA, Moghissi KS, Eliasson R, Paulsen CA, Callegos AJ: Laboratory Manual for the Examination of Human Semen and Semen-Cervical MucusInteraction, Singapore: Press concern; 1980

Gebert C, Wrenzycki C, Herrmann D, Gröger D, Reinhardt R: The bovine IGF2 gene is differentially methylated in oocyte and sperm DNA. Genomics 2006, 88:222–229