Volume 30, Issue 3 (5-2023)                   RJMS 2023, 30(3): 23-35 | Back to browse issues page

Ethics code: IR.SSRC.REC.1399.080

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kousha M, Abednatanzi H, Gholami M, Ghazalian F. The Effect of High-Intensity Interval Training and Thyme Honey Intervention on Insulin Resistance Index and the Expression of GATA4 in the Heart Tissue of Type 2 Male Diabetic Rats. RJMS 2023; 30 (3) :23-35
URL: http://rjms.iums.ac.ir/article-1-7267-en.html
Department of Physical Education and Sports Science, Science and Research Branch, Islamic Azad University, Tehran, Iran , abednazari@gmail.com
Abstract:   (691 Views)
Background & Aims: Diabetes mellitus is a disease that, is affected by environment and also genetics. The pathophysiological changes of diabetes are caused by dysfunction of peripheral β cells and ROS-dependent inflammation produced by tissues, all of these factors underlie insulin resistance and chronic inflammation, which gradually impairs blood glucose control (1). GATA4 is a cardiac-enriched transcription factor that is essential for various physiological and adaptive responses of cardiomyocytes. Recent studies have shown that GATA4 was able to protect cardiomyocytes from DOX-induced apoptosis, indicating that GATA4 also conveys a survival signal for cardiomyocytes (4). Recent studies have shown that physical activity has a significant effect on improving type 2 diabetes, reducing the risk of insulin resistance, metabolic syndrome and type 2 diabetes, and improving insulin sensitivity (8). Many studies show that by consuming more energy and training intensities, including high-intensity intermittent exercise, there is a greater response to insulin sensitivity throughout the body. During exercise, with muscle contraction and increased AMPK activity, stimulation of insulin sensitivity increases. After exercise, an increase in Akt inactivates TCB1D4 and thus increases the transport of glucose transporter (GLUT4) to the cell membrane, resulting Increases in absorption glucose (9). In recent years, in addition to the emphasis on exercise and physical activity, attention has been paid to dietary supplements and some herbs for the treatment of diabetes, among which there is great interest in the use of honey (15). Honey is primarily rich in carbohydrates and is also abundant in flavonoids and phenolic acids; thus, it is a promising therapeutic antioxidant for various disorders. it is a promising antidiabetic agent.  Although the use of honey due to its therapeutic and nutritional value has long been considered and approved in scientific texts, but its use in modern medical sciences is debatable (17). Therefore, considering the known physiological positive effects and protection of the heart, periodic exercise and thyme honey, as well as their positive role in improving cardiac function and the possibility of stimulating GATA4 gene expression in line with positive physiological adaptations of the heart, especially cardiomyocyte hypertrophy, we hypothesized that: 1- HIIT, Thyme honey  and The combination of both can be effective in increasing the expression of GATA4 gene in male type 2 diabetic rats. Therefore, the aim of the present study was to investigate the effects of eight weeks of HIIT and thyme honey on GATA4 gene expression in male type 2 diabetic rats.
Methods: In this experimental study, 36 young male Wistar rats, as a statistical sample, were purchased from Royan Research Institute and were move to the Razi Laboratory of Azad University of Science and Research.  After two weeks of getting acquainted with the laboratory environment and reaching a weight of 197± 20, undergo a high-fat diet (HFD) for 20 weeks on a high-fat diet prepared by the Royan Biotechnology Research Institute, which 45% HFD was given for 3 months and 60% HFD was given for 2 months (20, 21). Then Rats were randomly divided into four groups: control (n=8), HIIT (n=10), thyme honey (n=8) and HIIT*thyme honey (n=10) groups. At the end of the protocol, 28 rats were survived. In each group 2 rats died. To induce diabetes, a high-fat diet is used for 20 weeks and then intraperitoneal injection (25 mg / kg) of STZ solution is used. The HIIT training intervention of 8 weeks, with 2 min running at 80-90% VO2max and 1 min at 50-56% VO2max, was performed for 5 sessions per week including 2 to 8 intervals 16 Up to 34 minutes was done running on a treadmill, so that running time increased from 16 minutes in the first week to 34 minutes in last week. Additionally, during the 8 weeks of intervention, 3 g/kg of diluted thyme honey consumed, 5 days/week in supplement groups, honey and HIIT*honey groups.
Cardiac tissue was isolated and transferred to a negative 80 freezer to measure the expression of genes. In the next step, the RNA is extracted by RiboEX Total RNA isolation solution (GeneAll) and finally the quantitative and qualitative study of the RNA by Nanodrop device is evaluated. After ensuring the quality of RNA, it was extracted, cDNA made using the TAKARA kit and transferred to negative 20 freezer. AMPLIQON Master Mix Cybergreen was used for real-time reverse transcription-polymerase chain reaction (real-time RT-PCR). In the present study, the GAPDH reference gene was used as a house keeping gene and the Delta Delta CT method was used to quantitatively analyze the Real Time PCR data. The expression of the GATA4 genes in heart tissue. The primers used in this study were designed by Allel ID6 software and synthesized by Sinaclon Biotechnology Company. The sequence of primers is given in Table 2. Temperature gradient was set. All samples were repeated 2 times in a real-time PCR machine. Also, the temperature and time program of Real Time PCR reaction in the present study is according to Table 3. Data were analyzed using SPSS22 software. Shapiro-Wilk test to determine the normality of data distribution and Levin test for homogeneity of variances and inferential statistics of one-way analysis of variance and tukey post hoc test were used to compare the differences between groups. Significance level in the tests was considered α 05 0.05.
Results: According to the results of descriptive statistics, the scores of HIIT in the insulin resistance index are lower than the other groups. Also, the GATA4 scores in the HIIT and Theym honey are higher than the other groups.  ANOVA Analysis revealed that there was a significant difference between the groups in the variable of HOMA-IR (P = 0.001) and GATA4 gene expression (P = 0.016).The results of Tukey post hoc test showed that HOMA-IR was significantly decreased only in HIIT group when compared with other groups, whereas this finding was not repeated in other intervention groups (p>0.05).and expression of GATA4 gene was significantly increased in HIIT and Theym honey (p <0.05).
Conclusion: The aim of this study was to evaluate the effect of HIIT and thyme honey on insulin resistance index and expression of GATA4 gene in heart tissue of type 2 male diabetic rats. The results of the present study revealed that HOMA-IR were significantly decrease on the HIIT group when compared with others groups, though, there were no significant difference on the insulin resistance index  in thyme honey and  HIIT*thyme honey groups. These results were consistent with the study of Nova et al. (2017), Farazmandi and Rezaian (1399), Jelstad et al. (2021). also, Su et al. (2011) showed that exercise and physical activity can significantly reduce the insulin resistance index (34). Azimi Dokht et al. (2015) also showed in a study that eight weeks of intermittent exercise has a positive effect on fat profile and insulin resistance in type 2 diabetic men (35). Although, in concern with expression of GATA4 gene there are significantly increase in HIIT group in compared others group, there is no significant difference in the other intervention groups. These results are consistent with the findings of Hemmati et al. (2019), Xu et al. (2018), Naderi et al. (2019), Xiao et al. (2014). Xu et al. (2018) demonstrated GATA4 had a protective effect against endothelial dysfunction induced by hyperglycemia. Overexpression of GATA4 was demonstrated to lead to increased expression of NOX4 mRNA and protein. Furthermore, GATA4 overexpression resulted in increased nitric oxide (NO) production through the upregulation of endothelial NO synthase phosphorylation (38). Exercise imposes a higher cardiac workload, inducing angiogenesis and hypertrophy of cardiomyocytes, and GATA4 triggers this physiological process. However, overexpression of GATA4 is also associated with decompensated cardiac hypertrophy, as it occurs in neuroendocrine overactivation-induced HF. On the other hand, in postnatal hearts, GATA4 is essential for cardiomyocyte survival after injury or stress (38).
For years, people have believed that diabetics cannot consume honey because honey is high in sugar. Regarding honey, it should be noted that according to the standard, the acceptable limit for the ratio of fructose to glucose of honey is a minimum of 0.9 (40).Due to the fact that the glycemic index of fructose is very low. These findings show that honey causes fewer changes in blood sugar compared to glucose and can prevent the adverse effects of postprandial hypertension (41) and therefore this ability It increases the expression of GATA4 gene. Therefore, it seems that interval exercise and consumption of thyme honey can be used in type 2 diabetes.
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Type of Study: Research | Subject: Exercise Physiology

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