Page 60 - GTM-3-1

Global Translational Medicine Reproductive hormones and muscle in obesity

by decreased levels of testosterone, luteinizing hormone and No:2 on February 2, 2022) and was conducted in
(LH), and sex hormone-binding globulin. In contrast, accordance with the guidelines set forth by the European
2
obesity affects the health and metabolism of females and Convention for the Protection of Vertebrate Animals
3
their offspring. Along with reproductive hormones, (ETS No. 123).
corticosterone plays an important role in the pathogenesis The rats were euthanized through decapitation with prior
of obesity and MS. Notably, obesity also affects the anesthesia (sodium thiopental). Female rats were euthanized
4
condition of muscle tissues and can lead to progressive in the diestrus phase of the estrous cycle, determined by the
skeletal muscle atrophy and sarcopenia, as well as type of cells present in the rat vaginal swab. 10
decreased muscle mass and strength. MS may be caused
5,6
by decreased levels of anabolic hormones and increased The body weight of the rats was measured on a weighing
oxidative stress, correlating to MS and mitochondrial scale (Saturn ST-KS7230, China). After euthanasia, the
dysfunction in obesity. Mitochondrial dysfunction blood and tissues (i.e., visceral fat and musculus triceps
5
by oxidative stress results in excessive reactive oxygen surae) of the rats were collected and weighed on a
species (ROS) generation, and the overproduction of ROS laboratory weighing scale (Scout Pro, China). For the male
(exceeding the cellular antioxidant defense) damages the rats, the visceral fat mass included the paranephral and
cellular macromolecules and affects cellular functions and epididymal fat deposits. For the female rats, the visceral
viability. Given the multifaceted effects of obesity, it is of fat mass included the paranephral and periovarian fat
7
particular interest to further investigate these effects in deposits. Mass coefficients (MCs) of the organs and tissues
relation to gender, reproductive hormone levels, muscle were calculated using the following formula:
tissue condition, and their combined association. 8 MC = (Organ mass/body weight) ×100% (I)

Therefore, this study aimed to investigate the effects 2.2. Biochemical and hormonal parameters
of HCD on the development and progression of obesity,
reproductive hormone levels, and morphofunctional Biochemical parameters were measured from the serum
characteristics in the muscle tissues of both male and and muscle tissue homogenates of rats (1/10 dilution in
female Wistar rats. 0.1 M sodium phosphate buffer, pH 7.4) on a biochemical
automatic analyzer BS-200 (SHENZHEN MINDRAY
2. Methods Bio-Medical Electronics Co., LTD., China) and with
commercial kits, including DiaSens (Production unitary
2.1. Animals and diets enterprise “Diasens,” Republic of Belarus) and Lactat-
The study was conducted on 2-month-old sexually mature Vital (Vital Development Corporation, Russia). Quality
male and female Wistar rats. These rats were bred in-house control was maintained using commercial control sera
in a certified vivarium at the Institute of Physiology of the (Randox Laboratories Ltd., UK). Testosterone, estradiol,
National Academy of Sciences of Belarus. The rats were corticosterone, and LH levels were determined from the
kept under a 12/12 h light/dark cycle at a temperature of serum using Hema commercial kits (Xema Co., Russia).
22 ± 2℃ and humidity of 60–65%. The male (n = 27) and Testosterone, estradiol, and malondialdehyde (MDA) levels
female (n = 27) Wistar rats were randomly divided into two were also determined from muscle tissue homogenates
experimental groups: control and HCD. The control group, (1/10 dilution in 0.1 M sodium phosphate buffer, pH 7.4).
consisting of 13 male and 14 female Wistar rats, received MDA and superoxide dismutase (SOD) activities were
the standard diet (StD). The HCD group, comprising determined spectrophotometrically through reaction
14 male and 13 female Wistar rats, was given an HCD for with thiobarbituric acid and the inhibition of adrenaline
11
12
16 weeks. autooxidation, respectively.
The HCD consisted of StD supplemented with animal 2.3. Morphological analysis
fats (lard) (45% daily caloric content) and 10% fructose For morphological analysis, fragments of the musculus
solution (instead of water) ad libitum. The caloric content triceps surae were subjected to rapid freezing in a cryostat.
9
of StD for each rat was 150 kcal/day (i.e., the normal diet at Several 7 μm thick slices were made using the HM525
the vivarium of the Institute of Physiology of the National Cryostat (MICROM International GmbH, Germany)
Academy of Sciences of Belarus). In contrast, the caloric and stained with hematoxylin and eosin. The slides were
content of HCD for each rat was 228 kcal/day.
examined using the LUM-1 light microscope equipped
This study was approved by the Bioethics Committee with a digital camera (Altami LLC, Russia). The ImageJ
of the Institute of Physiology of the National Academy of software was used for morphometric analysis. On the
Sciences of Belarus (protocols No.:1 on January 22, 2021, digitized images, eight to ten transversely cut fibers were

Volume 3 Issue 1 (2024) 2 https://doi.org/10.36922/gtm.2321