Abstract

Cannabis sativa is a cheap hallucinating agent used in different parts of the world from time unknown as a part of various religious as well as social practices. Cannabis which is a special type of Marijuana can provide temporary relief from analgesia, body pain, and in some other clinical conditions. But impacts of Cannabis on reproductive health of males and females are multi-faceted and differentially fatal. In males, Cannabis can cause changes in testicular morphology, sperm parameters (in terms of semen quality, sperm morphology, sperm mortality, and sperm motility), male reproductive hormones and finally causing reduced libido. In females, Cannabis can reduce female fertility by disrupting hypothalamic release of gonadotropin releasing hormone (GnRH), leading to reduced estrogen and progesterone production and anovulatory menstrual cycles. Current research suggest that Cannabis may negatively impact on male and female fertility conditions. However, male sterility considering the Cannabis impact is totally lacking in human as well as in sub-human primates. However, very limited studies are available on Cannabis effect on primate female reproduction considering Rhesus monkeys. Hence, further studies are needed to validate that robust findings in animal models will carry over into human experience.

Highlights

  • Cannabis which is a type of marijuana has been used by the people of Indian subcontinent from time unknown [1]

  • All the above mentioned reported phenomenon are occurring in the central nervous system and no definitive proof has been reported till date how the enocannabinoids are affecting peripheral reproductive performances in females

  • Group 1 was treated with distilled water; group 2 was gavaged with 6 mg/100 g body weight/day aqueous Cannabis preparation; group 3 was gavaged with 12 mg/100 g bodyweight/ day aqueous Cannabis preparation

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Summary

Introduction

Cannabis which is a type of marijuana has been used by the people of Indian subcontinent from time unknown [1] They use this herb as a part of holy practice and use it for recreational purposes [2]. The main causative agent of marijuana/cannabinoids is the endocannabinoid This is a neutral lipid and highly conserved molecule throughout evolutionary history [11]. There are literatures suggesting the role of this ∆9-tetrahydrocannabinol (THC) in regulation of functions of central nervous system and regulating the reproductive functions by affecting/ modulating hypothalamo-pituitary-gonadal axis (HPG-axis) [16] via its receptor CB1 and CB2 [17]. All the above mentioned reported phenomenon are occurring in the central nervous system and no definitive proof has been reported till date how the enocannabinoids are affecting peripheral reproductive performances in females (in terms of gonadal activity, steroidogenesis, receptor expressions, free radical generations). Aim of the present study was to note the cannabinoid ( endocannabinoid) induced oxidative stress and reproductive impairments in female mice taking peripheral reproductive organs (ovary) in consideration

Animals and maintenance
Purity assessment of Cannabis preparations
Experimental design
Collection of desired tissues
Antibodies and reagents
Histological preparations
Immunohistochemistry of CB1 receptor
Estimation of total serum cholesterol
Evaluation of SOD activity in ovary
Estimation of catalase activity in ovary
Glutathione peroxidase (GPx) estimation in ovary
Caspase 3 activity assay
3.11 Western blot analysis of Cannabinoid receptor 1 (CB1) analysis
3.12 Statistical analyses
Histomorphology of ovary
Immunohistochemistry of CB1 receptor in ovary
Ovarian weight
Total serum cholesterol
SOD activity in ovary
4.11 Malondialdehyde level in ovary
4.13 Caspase 3 activities in ovarian thecal cells
4.14 Serum level of estradiol
4.15 Western Blot analysis of CB1 receptor in ovaries of mice
Discussions
Conclusion

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