Open Access, Peer-reviewed
pISSN 2288-6478
eISSN 2288-6761
    J Menopausal Med. 2023 Apr;29(1):29-39. English.
    Published online Apr 28, 2023.
    https://doi.org/10.6118/jmm.22037
    Copyright © by The Korean Society of Menopause
    Original Article

    Metabotropic Effect of Probiotic Supplementation and High-Intensity Interval Training in Menopause-Induced Metabolic Syndrome in Rats

    Zeinab Bayat,1,2 Arsalan Damirchi,1 Meysam Hasannejad-Bibalan,3 and Parvin Babaei2,4,5
      • 1Department of Exercise Physiology, Faculty of Physical Education and Sport Sciences, University of Guilan, Rasht, Iran.
      • 2Cellular and Molecular Research Center, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
      • 3Department of Medical Microbiology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
      • 4Neuroscience Research Center, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
      • 5Department of Physiology, School of Medicine, Guilan University of Medical Sciences, Rasht, Iran.
    • Address for Correspondence: Parvin Babaei, Cellular and Molecular Research Center, School of Medicine, Guilan University of Medical Sciences, 6th Km of Rasht-Tehran road, Guilan University Campus, Rasht 3363, Iran. Tel: 98-1333690884, Email: p_babaei@gums.ac.ir
    Received November 09, 2022; Revised March 14, 2023; Accepted March 20, 2023.

    This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/).

    Abstract

    Objectives

    This study aimed to investigate the interactive effect of supplementation with a native potential probiotic lactobacillus and 8-week high-intensity interval training (HIIT) on insulin resistance and dyslipidemia in a menopause-induced metabolic syndrome.

    Methods

    A total of 40 ovariectomized (OVX) Wistar rats were divided into five groups: control (OVX + Vehicle), exercise (EXE) (OVX + Exe), probiotic (Prob) (OVX + Prob), exercise and probiotic (OVX + Exe + Prob), and sham surgery. After the end of the treatment interventions, body weight, body mass index (BMI), waist circumference (WC), visceral fat, and serum concentrations of glucose, insulin, lipid profile, and adiponectin were measured using colorimetric analysis and enzyme-linked immunosorbent assay, respectively.

    Results

    Data revealed a significant decrease in weight, waist circumference , visceral fat, BMI, and levels of glucose, insulin, homeostasis model assessment of insulin resistance, triacylglyceride, total cholesterol, and low-density lipoprotein (LDL), but an increase in high-density lipoprotein and adiponectin levels (P = 0.001), in OVX + Exe + Prob compared with the OVX + Vehicle group.

    Conclusions

    The present study indicates that native probiotic lactobacillus combined with HIIT effectively reduces body weight, visceral fat, and levels of LDL, glucose, and insulin and increases adiponectin level, although exercise contributes more to fat reduction and probiotics to insulin resistance.

    Keywords
    Adiponectin; Insulin resistance; Lipids; Metabolic syndrome; Probiotics

    INTRODUCTION

    Obesity is a major risk factor for various metabolic and cardiovascular diseases (CVD) [1], and includes a complex metabolic cluster such as metabolic syndrome (MetS), insulin resistance (IR), and hypertension [2]. MetS is defined by at least three of five indices according to the world Health Organization: central obesity, hypertension, insulin resistance, and atherogenic dyslipidemia [3]. The worldwide prevalence of MetS is one in four adults [4], which raise up to one in three among post-menopausal women [2]. MetS increases the risk of developing CVD and type 2 diabetes [5, 6]. It is believed that, estrogen deficiency in menopausal women leads in weight gain, redistribution of fat, hyperlipidemia and high blood sugar [2, 7, 8], thus predisposing them to MetS, even independently of aging, as it is evident in premature menopause [9].

    Considering the improvement in hygiene, women spend at least one-third of their life in a menopausal period, therefore, finding safe treatments to prevent the progressions of MetS has great importance [10]. One of the safe and available strategies to combat with MetS is physical activity [11], particularly high intensity interval training (HIIT), which implies much more health benefits compared with low and mild intensities [12, 13]. Compared with moderate-intensity continuous training, HIIT has been reported to more effectively increases aerobic capacity (maximal oxygen consumption [VO2max]) and reduces risk factors associated with MetS, including blood pressure, and dyslipidemia [14]. However, despite all beneficial effects of exercise training, it has been questionable why some individuals do not significantly response to exercise protocols [15, 16]. We here assumed that individual’s microbiome might be one possible reason.

    Recently, modulation of gut microbiome has got attention in MetS [17], because intestinal microbiota has been known as a key player in the development of a chronic low-grade inflammatory state associated with MetS [17], and probiotics administration has come to the forefront to modulate intestinal microbiota [18]. Probiotic are live microorganisms, mostly the strains of lactic acid bacteria (Lactobacillus spp) and Bifidobacterium spp which are administered in adequate amounts to confer a health benefit in the host [19, 20]. These microorganisms contribute in various physiological functions including immune system improvement, vitamins and short-chain fatty acids synthesis, homeostasis of digestive tract [21] and alleviation of IR [17].

    The core component for MetS is adipose tissue, which releases specific peptides named adipokines. Adipokines consist of pro and anti-inflammatory cytokines and contribute to various physiological functions [22]. Among various adipokines, adiponectin is an anti-inflammatory adipocytokine that plays a crucial role in glucose and lipid metabolism [23], insulin resistance, and atherosclerosis [22]. While low level of adiponectin is associated with IR [22, 24], the increased adiponectin induces insulin sensitivity in skeletal muscles by binding with adiponectin receptor type 1 and 2, and leading to inhibition of various inflammatory cytokines [22]. Currently, adiponectin stands as anti-inflammatory and anti-obesity adipokine [2].

    Therefore, in the present study, we hypothesized that consumption of native potential probiotic bacteria together with HIIT might efficiently alleviate MetS indices. To approach this, we induced menopausal standard model of MetS with ovariectomy surgery, then the animals underwent the related treatments for eight weeks and the related morphometric and biochemical factors were evaluated.

    MATERIALS AND METHODS

    Animals

    Forty adult female Wistar rats (3 months age and 180–200 g weight) were housed four per each cage in a room with a temperature of 22℃ ± 2℃, and 12/12-hour light/dark cycle (light on 07:00) with free access to food and water [25]. Animals fed normal diet which described in Table 1.

    Table 1
    Ingredients of rodents standard diet per 100 g

    Metabolic syndrome was induced by ovariectomy surgery, which is an appropriate model to increase fat mass, waist circumference and IR [26, 27]. Experimental methods were conducted according to the EU animal experiments ethical guidelines (2010/63/EU) and got an approval code from the Sport Sciences Research Institute of Iran (code IR.SSRI.REC.1400.1087).

    Ovariectomy surgery

    Rats were ovariectomized under general anesthesia with an intraperitoneal (i.p) injection of ketamine (50 mg/kg) and xylazine (5 mg/kg) in a ratio of 4:1 [27], and then ovaries were removed through single midline incision according to our previous study [28]. The sham surgery (sham) group had the surgical procedure without removal of ovaries.

    Groups and intervention protocol

    One month after ovariectomy, animals were randomly divided into the following groups (n = 8): exercise (ovariectomized [OVX] + exercise [Exe]), probiotic (OVX + probiotic [Prob]), and combination of exercise with probiotic (OVX + Exe + Prob), vehicle group (OVX + Vehicle). Exercised groups ran on a five lines rodent treadmill (DSI-580; Danesh Salar Iranian) with intensity of 90% to 95% VO2max and zero-degree gradient for three sessions/week for eight weeks. Probiotic groups received oral consumption (by gavage syringe) of 1 mL cocktail of Lactobacillus species consisting of 109cells, for three times per week. The control group (OVX + Vehicle), received the same volume of drink vehicle by gavage syringe. Design of the study was illustrated in Figure 1.

    Fig. 1
    Design of study. Animal adaptation to the new environment for 1 week; HIIT + Probiotic supplementation: high-intensity interval training (HIIT with intensity of 90% to 95% VO2max and zero-degree gradient for three sessions/week for 8 weeks), consumption probiotic (received oral consumption (by gavage syringe) of 1 mL cocktail of Lactobacillus species consisting of 109 cells, for three times per week for 8 weeks). Blood sampling was carried out 48 hours after the last exercise session. HIIT: high-intensity interval training, MetS: metabolic syndrome.

    Preparation of Lactobacillus strains

    According to the previous studies the Lactobacilli cocktail is more effective than a single strain, therefore, we used the Lactobacilli cocktail in the present study [29, 30]. Probiotic source was fecal samples provided from participatnts in our previous cross-sectional study conducted in 2019 (Ethics approval code: IR.GUMS. REC.1398.016) [31]. Eighty three volunteer individuals from the city of Rudbar which is located in the north of Iran participated in that study. Rudbar is a nonindustrial city with a humid and rainy climate, and vegetables are a major part of the diet of people in this area. Samples were taken from volunteer’s who were referred to rural health centers by convenience sampling method, and received written informed consent from participants for further research on isolated lactobacillus. Inclusion criteria were lack of gastrointestinal diseases and antibiotics consumption over six months’ period before sample collection. The probiotic properties of the employed Lactobacillus strains used in this work included bacteriocins production and bactericidal effects, tolerance to acid and bile [31]. In order to perform the test, An De Man, Rogosa, and Sharpe (MRS) agar plate was streaked for isolation from the glycerol bacterial stocks and incubated the plates at 37℃ for 48 hours to ensure purity. A single colony was cultured in MRS broth and after 48 hours, the final density of strains at 109 CFU per 1 mL was prepared. finally. each rat received 1 mL of 109 colony forming units/mL (three times per week through the gavage syringe of Lactobacilli cocktail) [32].

    HIIT program

    Animals were placed on a treadmil and ran at a speed of 10 m per minute for 10 minutes with a zero-degree gradient for one week in order to acclimate to apparatus and stress. Then two groups of HIIT ran on treadmill three sessions per week for 8 weeks with the intensity of 90% to 95% VO2max and 9 times of one-minute intervals consisting of running with the intensity of 50% VO2max (Fig. 2). For warming up and cooling down animals ran 4 minutes with an intensity of 55% VO2max as well (Fig. 2). To measure VO2max, rats ran on the treadmill at a speed of 6 m/min with a zero-degree gradient for 5 minutes, and then every 3 minutes the speed of the treadmill, rises to 3 m/min until the animals reach the level of exhaustion [33].

    Fig. 2
    HIIT protocol includes warming up and cooling down, 4 minutes with intensity of 55% VO2max. HIIT: 9 one-minute intervals with intensity of 90% to 95% VO2max and one-minute with the intensity of 50% VO2max between intervals with zero-degree gradient for three sessions per week for 8 weeks. VO2max: maximal oxygen consumption, HIIT: high-intensity interval training.

    Measurement of visceral fat, biochemical and morphometric parameters

    Morphometric parameters measurement

    All rats were weighed weekly, between 09:00 a.m. and 11:30 a.m. using a precise digital scale (Jadever Scale Co) with an accuracy of 0.1 g. After one night fasting, rats were deeply anesthetized and sacrificed for measuring weight, waist circumference (WC) and height [34]. WC was measured on the largest region of the rat's abdomen [28]. Then blood samples were drawn from the inferior vena cava, and serums were separated by rapid centrifuge (3,000 rpm for 15 minutes) and stored at –80℃ for later biochemical measurements. After collecting the blood samples, all intra-abdominal fat depots including mesenteric, urogenital and retroperitoneal were dissected out. Mesenteric fat pad consisted of adipose tissue surrounding the gastrointestinal tract from the gastroesophageal sphincter to the end of the rectum. Urogenital fat pad included adipose tissue surrounding the kidneys, ureters, and bladder as well as ovaries, oviducts, and uterus. The retroperitoneal fat pad was taken distinctly behind each kidney along the lumbar muscles and weighed immediately after dissection using lab weighing scale (SKY-3000; Jadevar, with an accuracy of 1 mg) to avoid evaporative weight loss [34].

    Biochemical measurements

    Serum adiponectin and insulin were measured by enzyme-linked immunosorbent assay (ELISA) using Mouse Adiponectin/Acrp30 DuoSet ELISA (Bio-Techne) Rat / Mouse Insulin ELISA Kit (Creative Diagnostic®). Serum glucose, total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) concentrations were determined by enzymatic colorimetric method (GOD-PAP; Pars Azmoun). Triglyceride (TG) was assessed by Triglyceride Quantification Kit, Catalog Number MAK266 (Pars Azmoun). The procedure in Babaei et al. [35] was used to estimate low-density lipoprotein cholesterol (LDL-C).

    Finally, IR index (IRI) was assessed by homeostasis model assessment estimate of IR (HOMA-IR) according to the following formula:

    Fasting insulin (µU/mL) × fasting glucose (mmol/L) / 22.5

    MetS severity Z-score was calculated by the following formula according to the previous studies [28]:

    [(HDL – 37) / 8.3] + [(LDL – 22)/3.9] + [(TG – 42) / 7.1] + [(fasting glucose – 130) / 15.5] + [(insulin – 7) / 2.6] + [(WC – 17) / 1.14]

    Statistical analysis

    Normality of data was estimated by Shapiro-Wilk test using SPSS software version 22.0 (IBM SPSS Co.). One-way analysis of variance (ANOVA) and post hoc Tukey tests were used for group comparisons. Results are expressed as mean ± SE of the mean and P < 0.05 was considered statistically significant.

    RESULTS

    Cotreatment of probiotic and HIIT decreased weight, BMI, waist circumference and visceral fat in menopausal model of rats with MetS

    At the end of the interventions, data analyzed by one-way ANOVA test showed significant differences between groups in body weight (F(4,35) = 34.100, P = 0.001), WC (F(4,35) = 58.778, P = 0.001), visceral fat (F(4,35) = 54.849, P = 0.001) and BMI (F(4,35) = 11.131, P = 0.001) (Fig. 3).

    Fig. 3
    The effect of exercise, probiotic, and combination of both on (A) body weight, (B) WC, (C) visceral fat, and (D) BMI. OVX + Vehicle: control group received of drink; OVX + Exe: HIIT group; OVX + Prob: consumption probiotic group; OVX + Exe + Prob: consumption probiotic with HIIT group. One-way analysis of variance test followed by Tukey’s post hoc test. Values are reported as the mean ± SE of the mean, n = 8 rats per group. OVX: ovariectomized, Exe: exercise, Prob: probiotic, WC: waist circumference, HIIT: high-intensity interval training, BMI: body mass index, Sham: sham surgery. **P ≤ 0.01, ***P ≤ 0.001 versus OVX + Vehicle group. #P ≤ 0.05, ###P ≤ 0.001 versus sham group. $P ≤ 0.05, $$P ≤ 0.01 versus OVX + Exe + Prob group.

    Tukey test pairwise comparisons showed 30% (P = 0.001) increase in weight in the group of OVX + Vehicle compared with Sham, but 12%, 9% (P = 0.001) and 7% (P = 0.004) reduction in OVX + Exe + Prob, OVX + Exe, and OVX + Prob respectively compared with OVX + Vehicle (Fig. 3A). There was no significant difference between monotherapies and cotreatment groups.

    Moreover, Tukey test showed a significant increase in WC of OVX + Vehicle (23%, P = 0.001) compared with Sham, which followed by 14%, 12% and 10% reduction in OVX + Exe + Prob, OVX + Exe, OVX + Prob (P = 0.001) compared with OVX + Vehicle. Also OVX + Exe + Prob group showed significant reduction compared with OVX + Prob (4%, P = 0.02), but not in comparison with the OVX + Exe group (Fig. 3B).

    Visceral fat was increased by 69% (P = 0.001) in OVX + Vehicle compared with Sham, and then was reduced by 31% in OVX + Exe + Prob, 25% OVX + Exe, and 22% in OVX + Prob in comparison with the OVX + Vehicle (P = 0.001). Also, OVX + Exe + Prob group showed significant reduction compared with OVX + Prob (12%, P = 0.01), but not with the OVX + Exe group (Fig. 3C).

    Finally, significant elevation in BMI was found in OVX + Vehicle (19%, P = 0.001) compared wtih Sham, while it was reduced by 13% in OVX + Exe + Prob (P = 0.001), and 10% in OVX + Exe and OVX + Prob (P = 0.002) compared with the OVX + Vehicle. There was no significant difference between treated groups (Fig. 3D).

    Cotreatment of probiotic and HIIT decreased serum glucose and insulin in menopausal model of rats with MetS

    A significant difference was found in glucose (F(4,35) = 11.771, P = 0.001), insulin (F(4,35) = 7.750, P = 0.001), and HOMA-IR (F(4,35) = 13.365) (Fig. 4). Glucose was increased in OVX + Vehicle by 27% (P = 0.001) compared with Sham, and decreased by 23% in OVX + Exe + Prob (P = 0.001), 15% and 18% in OVX + Exe (P = 0.002) and OVX + Prob (P = 0.001) respectively compared with the OVX + Vehicle group. There was no significant difference between these three groups (Fig. 4A).

    Fig. 4
    The effect of exercise, probiotic, and combination of both on (A) glucose, (B) insulin, (C) HOMA-IR. OVX + Vehicle: control group received of drink: OVX + Exe, HIIT group; OVX + Prob: consumption probiotic group; OVX + Exe + Prob: consumption probiotic with HIIT group. One-way analysis of variance test followed by Tukey’s post hoc test. Values are reported as the mean ± SE of the mean, n = 8 rats per group. HOMA-IR: homeostasis model assessment of insulin resistance, OVX: ovariectomized, Exe: exercise, Prob: probiotic, HIIT: high-intensity interval training, Sham: sham surgery. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 versus OVX + Vehicle group. ###P ≤ 0.001 versus sham group. $P ≤ 0.05 versus OVX + Exe + Prob group.

    Also insulin significantly was increased in OVX + Vehicle (48%, P = 0.001) compared with Sham, and decreased by 49%, in OVX + Exe + Prob (P = 0.001), 20% in OVX + Exe (P = 0.02), and 29% in OVX + Prob (P = 0.003) compared with the OVX + Vehicle group. OVX + Exe + Prob group showed 35% reduction in insulin compared with OVX + Exe (P = 0.03), but no significant difference with OVX + Prob group (Fig. 4B).

    Finally 89% elevation in HOMA-IR was found in OVX + Vehicle (P = 0.001) compared wtih Sham, while it was reduced by 60% in OVX + Exe + Prob (P = 0.001), 32% in OVX + Exe (P = 0.007), OVX + Prob (41%, P = 0.001) compared with the OVX + Vehicle group. Also OVX + Exe + Prob group showed 40% reduction compared with OVX + Exe (P = 0.02), but not significant difference with OVX + Prob group (Fig. 4C).

    Cotreatment of HIIT and probiotic modulated lipid profile in menopausal model of rats with MetS

    One-way ANOVA test showed significant differences between groups in TG (F(4,35) = 18.286, P = 0.001), TC (F(4,35) = 8.455, P = 0.001), HDL (F(4,35) = 32.129, P = 0.001), LDL (F(4,35) = 10.721, P = 0.001), and very low density lipoprotein (VLDL) (F(4,35) = 6.110, P = 0.001) (Fig. 5).

    Fig. 5
    The effect of exercise, probiotic, and combination of both on (A) TG, (B) TC, (C) HDL-C, (D) LDL-C, (E) VLDL. OVX + Vehicle: control group received of drink; OVX + Exe: HIIT group; OVX + Prob: consumption probiotic group; OVX + Exe + Prob: consumption probiotic with HIIT group. One-way analysis of variance test followed by Tukey’s post hoc test. Values are reported as the mean ± SE of the mean, n = 8 rats per group. TG: triacylglyceride, TC: total cholesterol, HDL-C: high-density lipoprotein cholesterol, LDL-C: low-density lipoprotein cholesterol, HIIT: high-intensity interval training, VLDL: very-low-density lipoprotein. *P ≤ 0.05, ***P ≤ 0.001 versus OVX + Vehicle group. #P ≤ 0.05, ###P ≤ 0.001 versus sham group.

    TG significantly was increased in OVX + Vehicle (43%, P = 0.001) compared with Sham group, but it was decreased in OVX + Exe + Prob by 27%, OVX + Exe (25%), OVX + Prob (17%) (P = 0.001) compared with OVX + Vehicle. There was no significant difference between these three groups (Fig. 5A).

    TC was raised in OVX + Vehicle by 18% (P = 0.001) compared with Sham group, and reduced by 16% in OVX + Exe + Prob, 13% in OVX + Exe (P = 0.001), OVX + Prob (11%, P = 0.01) compared with OVX + Vehicle group, but There was no significant difference between these three groups (Fig. 5B).

    HDL was significantly decreased in OVX + Vehicle (24%, P = 0.001) compared with Sham group, but elevated by 42% in OVX + Exe + Prob, 32% OVX + Exe, and 34% in OVX + Prob (P = 0.001). There was no significant difference between these groups (Fig. 5C).

    Also 45% increase in LDL belonged to the group of OVX + Vehicle (P = 0.001) compared with sham, whereas it was decreased by 33%–34% (P = 0.001) either by cotreatment of probiotic and HIIT or monotherapy of them, with no significant difference between them (Fig. 5D).

    Significant elevation in VLDL, also was found in OVX + Vehicle (20%, P = 0.001) compared with sham group. Then after treatments it was reduced by 18% in OVX + Exe + Prob and OVX + Exe (P = 0.001), and 10% in OVX + Prob (P = 0.02) compared with the OVX + Vehicle. No significant difference was found between these three groups (Fig. 5E).

    Cotreatment of HIIT and probiotic modulated serum adiponectin level in menopausal model of rats with MetS

    One-way ANOVA test showed significant differences between groups in serum adiponectin level (F(4,35) = 26.061, P = 0.001) (Fig. 6), and further Tukey test pairwise comparisons showed 18% decrease in OVX + Vehicle (P = 0.006) compared with sham group. Interestingly remarkable elevation in OVX + Exe + Prob (60%, P = 0.001), OVX + Exe (30%, P = 0.001), OVX + Prob (22%, P = 0.006) was found in comparison with the OVX + Vehicle group (Fig. 6).

    Fig. 6
    The effect of exercise, probiotic, and combination of both on Adiponectin. OVX + Vehicle: Control group received of drink; OVX + Exe: high-intensity interval training group; OVX + Prob: consumption probiotic group; OVX + Exe + Prob: consumption probiotic with high-intensity interval training group. One-way analysis of variance test followed by Tukey’s post hoc test. Values are reported as the mean ± SE of the mean, n = 8 rats per group. OVX: ovariectomized, Exe: exercise, Prob: probiotic, Sham: sham surgery. **P ≤ 0.01, ***P ≤ 0.001 versus OVX + Vehicle group. ##P ≤ 0.01 ###P ≤ 0.001 versus sham group. $$$P ≤ 0.001 versus OVX + Exe + Prob group.

    Finally, one-way ANOVA test showed a significant difference in MetS Z-score between groups (F(4,35) = 31.677, P = 0.001). Tukey test pairwise comparisons showed a significant increase in MetS Z-score of OVX + Vehicle (P = 0.001) compared with Sham and remarkable decrease in OVX + Exe + Prob, OVX + Exe, OVX + Prob groups compared with the OVX + Vehicle group (P = 0.001). No significant difference was found in MetS Z-score between the treated groups (Fig. 7).

    Fig. 7
    The effect of exercise, probiotic, and combination of both on MetS Z-score. OVX + Vehicle: Control group received of drink; OVX + Exe: HIIT group; OVX + Prob: consumption probiotic group; OVX + Exe + Prob; consumption probiotic with HIIT group. One-way analysis of variance test followed by Tukey’s post hoc test. Values are reported as the mean ± SE of the mean, n = 8 rats per group. MetS: metabolic syndrome, OVX: ovariectomized, Exe: exercise, Prob: probiotic, HIIT: high-intensity interval training, Sham: sham surgery. ***P ≤ 0.001 versus OVX + Vehicle group. ###P ≤ 0.001 versus sham group.

    DISCUSSION

    Our findings in line with the previous studies showed dyslipidemia [34, 36, 37], IR [28, 38], and increase in body weight, visceral fat, BMI and waist circumference [28, 34] in ovariectomized group compared with sham confirming the establishment of MetS model. Therefore, ovariectomy is an accepted model of menopause-induced metabolic syndrome [26].

    It has been previously reported that three weeks after ovariectomy, serum estrogen concentration is significantly decreased [27, 35], and this leads in glucose elevation and IR due to malfunction of translocation of GLUT4 [27]. Meanwhile, weight gain and visceral fat accumulation in our subjects, might be explained by adipocyte hypertrophy and reduction in lipase activity after ovariectomy [27, 34, 36]. Dyslipidemia observed here is justifiable, because estrogen has been known as an important hormone for activation of lipoprotein lipase, hormone-sensitive lipase, and also apolipoproteins for HDL [25]. In addition, change in microbiome after menopause may be also responsible for development of obesity via activating lipogenesis [39].

    In next part of the present study, both HIIT and probiotic supplementation per se, resulted in a significant decrease in visceral fat, BMI, WC, glucose, insulin, HOMA-IR, TG, TC, LDL, and increase in adiponectin and HDL compared with OVX group.

    Although combined of two treatments alleviated all IR indices, in parallel with 60% increase in adiponectin level, our present study showed no significant difference between monotherapy and cotreatment of HIIT and probiotic in MetS Z-score. These findings indicates that adding probiotic supplementation to HIIT significantly potentiates secretion of adiponectin, and adiponectin stimulated the shifting glucose into the target cells probably via 5’ AMP-activated protein kinase/phosphoinositide 3-kinases signaling pathway [27] or reduce blood lipids by peroxisome proliferator-activated receptor alpha [17, 23].

    Although previous studies showed that HIIT per se is capable to activates glucose transporters [37, 40], and also increase lipolysis [41], our data showed that exercise contributes more to waist circumference and fat mass reduction, whereas probiotic contributes more to insulin resistance. Probiotics have been known to exert have anti-inflammatory actions [39], and also capable to decrease body weight and fat mass through both peripheral mechanisms of modulating gut microbiota, and central mechanisms targeting satiety [42]. Because no significant difference was found between groups in terms of feeding, therefore peripheral mechanisms are more favorable. One probable mechanism for insulin sensitizing effects of probiotics in our study might be their potency to catabolize complex polysaccharides to short-chain fatty acids (SCFAs), to improve non-fat weight gain and glucose homeostasis [17]. Some of the SCFAs such as acetate suppresses insulin signaling in adipocytes, inhibits fat accumulation in adipose tissue, and decreases inflammatory state [17]. Meanwhile, improvement in gut barrier function by probiotics, reduces metabolic endotoxemia, and systemic inflammation, deconjugation of bile acids, which interfere with lipid absorption [43], and cholesterol assimilation, and removal [44].

    Taken all together, our findings suggest that combination of HIIT and local lactobacillus successfully regulates energy homeostasis, IR, and dyslipidemia.

    The strength of the present study is evaluating the effect of coadministration of native potential probiotic bacteria with a chronic protocol of HIIT on MetS indices and serum level of adiponectin. However, lack of probiotics dose response data and evidence for oxidative stress pathway alteration remain as limitations.

    In clinical importance of view, manipulation of the gut microbiota with lactobacillus supplementation together with a protocol of HIIT for eight weeks might be a safe strategy for alleviating MetS.

    Our findings suggest that chronic high intensity interval training combined with lactobacillus supplementation successfully alleviate some of the components of metabolic syndrome at least party via adiponectin dependent way.

    Notes

    FUNDING:This study was supported by grants from the Guilan University received by Dr Arsalan Damirchi.

    CONFLICT OF INTEREST:No potential conflict of interest relevant to this article was reported.

    ACKNOWLEDGMENTS

    The authors thank Hamid Morovatti and Shirin Javer for technical supports.

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    MeSH Terms
    Adiponectin
    Animals
    Body Mass Index
    Body Weight
    Cholesterol
    Dyslipidemias
    Enzyme-Linked Immunosorbent Assay
    Glucose
    High-Intensity Interval Training*
    Homeostasis
    Insulin
    Insulin Resistance
    Intra-Abdominal Fat
    Lactobacillus
    Lipids
    Lipoproteins, HDL
    Lipoproteins, LDL
    Metabolic Syndrome*
    Probiotics*
    Rats*
    Rats, Wistar
    Respect
    Waist Circumference
    Metrics
    Share
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