P–124 The beneficial effects of novel quercetin delivery system (loaded on bigel) on male fertility parameters of Non-alcoholic fatty liver model: an in-vivo (rat model) study
Authors: Hosseini E , Mousavi S N , Seye. Dorraj M S , Sheik. Mohammadi S , Pourmansoori Z , Rasulifard M H , Doosti M , Chiti H 
Published: August 2021

Abstract

Study question
Dose quercetin encapsulated in a bigel slow- release delivery system improve male fertility parameters in Non-alcoholic fatty liver (NAFLD) model

Summary answer
Quercetin in a bigel slow- release delivery system can boost semen parameters in NAFLD rat model

What is known already
Recent molecular and physiological studies have shown that adverse effects of NAFLD extend far beyond the liver. NAFLD can impair male reproductive function by increasing Reactive Oxidative Stress (ROS) levels, reducing the expression of antioxidant genes and inducing damage in testes immune privilege.
Antioxidant therapy and its effectiveness depend on whether the exogenous antioxidant will be readily absorbed to reach high enough that are required to decrease the pathological damages.
Quercetin, as an antioxidant, is able to ameliorate oxidative stress. The design of new drug delivery systems using encapsulating antioxidant can boost its durability and effectiveness.

Study design, size, duration
Bigels were prepared using cottonseed oil/cannabis oil/alginate/ferula gum. Sprague-Dawley rats are housed for 2 weeks, then NAFLD was induced by 58% of dietary calorie as lard and 42 g/L fructose for 16 weeks. The experimental protocol was approved by the ethical committee of Zanjan University of Medical Sciences, Zanjan, Iran.

Participants/materials, setting, methods
After confirming the NAFLD induction, animals divided into five groups: Control, control NAFLD, received 2 mg/kg Quer loaded on bigels, free bigels, free Quer for 45 days as daily gavage. Semen parameters (count, motility, and morphology), viability (Eosin-nigrosine staining) and serum testosterone levels were analyzed. In addition, histological sections of testicular tissues were investigated by Hematoxylin-Eosin (H&E) staining method. In situ detection of apoptosis was performed using terminal deoxynucleotidyl-transferase dUTP nick end labeling (TUNEL) assay.

Main results and the role of chance
The sperm count, sperm motility, normal morphology and testosterone level were significantly lower in the NAFLD group than those the controls. Moreover, higher head and tail abnormality percentages were seen in the sperm of these groups. Bigel-Quer significantly improved the serum testosterone level, sperm count, motility, and morphology compared with the NAFLD group. Spermatogenic cells in all stages of differentiation (spermatogonia, primary spermatocytes, early spermatids, late spermatids) are observed and preserved normally in the testicular tubules and lumen filled with mature sperms in the control group. Interestingly, atrophic changes in the testicular tubule architecture with swelling in spermatogonia cells, detachment from tubule membrane, reduced number of mature sperm, and reduced lumen thickness were seen in the NAFLD. In the Quer, bigel and bigel-Quer-treated groups, swelling and vacuolation rate of germ cells decreased. The testicular morphology, and tubule structure were significantly normalized, especially in the bigel-Quer-treated group. Serum testosterone levels significantly increased and reached the healthy control group in the bigel-Quer group. TUNEL-positive cells in testes increased significantly after NAFLD induction. Quantitative analysis showed a significant decrease in testicular TUNEL-positive cells following bigel-Quer treatment, but not in other groups.

Limitations, reasons for caution
Keeping and daily handling of animals for long-time in animal house for diet-induced NAFLD. NAFLD requires long periods of treatment to get the desired outcome especially in the case of sperm parameters investigation.
Wider implications of the findings: The bigel showed synergistic effects with Quer for treating infertility in male rats with NAFLD. Stability and bio-availability of Quer are important aspects that should be considered to justify its supplementation. Empowering antioxidant shield of NAFLD patients by Quer supplementation can improve various damage effects and clinical status of diseases.

Trial registration number
Not applicable