The Effect of Eight-Weeks of Aerobic Training with Nano-eugenol Supplementation on Pancreatic Tissue UPC3 Gene Expression and MDA Serum Levels in Diabetic Rats

Subject Areas : Open Access

Shirin  Fotovat ¹ (Department of Sport Physiology, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran)
Khosro  Jalali dehkordi ² (Department of Sport Physiology, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran)
Farzaneh  Taghian ³ (Department of Sport Physiology, Isfahan (Khorasgan) Branch, Islamic Azad University, Isfahan, Iran)

Received: 2020-12-26 Accepted : 2020-12-26 Published : 2020-11-21

Keywords: Aerobic training, Malondialdehyde, Diabetes, UPC3, Nano Eugenol,

Abstract :

Abstract Introduction: The use of exercise and herbal supplements to treat diseases and metabolic disorders has become popular among the people. Regarding the health-enhancing effects of aerobic training and eugenol, the present study aimed to investigate the effect of eight weeks of aerobic training with nano-eugenol supplementation on the gene expression of non-placental protein 3 (UPC3) in pancreatic tissue and serum malondialdehyde (MDA) levels in diabetic rats. Materials and Methods: In this experimental study, 20 diabetic rats were divided into four groups of five rats: 1) control, 2) aerobic training, 3) nano-eugenol, and 4) nano-eugenol + training. To investigate the effects of diabetes induction on research variables, five healthy rats were placed in a healthy control group. For eight weeks, groups 3 and 4 received 2 mmol of nano-eugenol by gavage daily, and groups 2 and 4 performed aerobic training at 30 m/s five days a week. Kolmogorov-Smirnov tests, one-way analysis of variance with Tukey post- hoc test were used to analyze the data (P≤0.05). Findings: Induction of diabetes had a significant effect on increasing UPC3 gene expression in pancreatic tissue and MDA serum levels (P=0.001). However, aerobic training with nano-eugenol had a significant effect on the reduction of UPC3 gene expression in pancreatic tissue (P=0.001) and serum levels (P=0.01). Conclusion: It seems that aerobic training along with the use of nano-eugenol has more favorable effects on improving UPC3 gene expression levels and MDA serum levels than the either one alone in diabetes. Keywords: Aerobic Training, Nano Eugenol, UPC3, Malondialdehyde, Diabetes   Extended Abstract Introduction Type 2 diabetes mellitus is a common and growing global health issue. According to the American Diabetes Association, there are about 400 million people with diabetes in the world and it is expected to be increased to more than 600 million people in 2035. In recent years, exercise has been used as a non-pharmacological strategy to control diabetes and its complications. Exercise improves inflammatory diseases such as diabetes by reducing the secretion of inflammatory cytokines and increasing anti-inflammatory cytokines (3). Unpaired protein 3 (UPC3) catalyzes adaptive calorific value in muscle due to the increase of proton transfer from the inner mitochondrial membrane. Numerous studies have shown decreased expression of UPC3 mRNA and UPC3 protein in the skeletal muscle of subjects or as a result of aerobic training (4). Currently, the main effective treatment for diabetes is the use of insulin and synthetic drugs that lower blood glucose; but these compounds have several side effects. Given that medicinal plants have fewer side effects than synthetic drugs, therefore, researchers are looking for herbal compounds to treat or prevent this disease. Eugenolor 4-ally1-2-methoxyphenol is an aromatic molecule that is textured in the oils and in the essential oils of various plants, including cloves and cinnamon (6). Various studies have shown that natural antioxidants with phenolic structure play an important role in protecting body tissues against free radicals. According to the presented evidence, eugenol has an important protective effect against lipid peroxidation originated from free radical (7). Therefore, the present study was done to shed light on the ambiguities in this field due to the increasing prevalence of diabetes and its harmful effects on health and the incidence of its complications as well as the effect of antioxidant supplements and the specific type of physical activity on its control and lack of studies on the interactive effects of aerobic training and the use of nano-eugenolon the expression of UPC3 gene and MDA serum levels in diabetic patient.   Materials and Methods In this experimental study, 25 male Wistar rats were purchased and transferred to the laboratory. Subsequently, 20 rats underwent intraperitoneal injection of streptozotocin (STZ). After confirming the rats as diabetic, fasting glucose-based diabetic rats were divided into four groups of five series including 1) training, 2) eugenol, 3) training + eugenol and 4) control. To investigate the effects of diabetes induction on research variables, the remaining five rats were placed in a healthy control group. For eight weeks, groups 3 and 4 received 2 mmol nano-eugenol daily by gavage (10) and groups 2 and 4 performed aerobic training at a speed of 30 m/min, five days a week according to Table 1 (11). It is noteworthy that eugenol was purchased specifically from Sigma and dissolved in dimethyl sulfoxide (DMSO). Nanoparticles were prepared according to the protocol with a concentration of 10% (10). Forty-eight hours after the last training session and after 14 hours of fasting, rats were injected intraperitoneally with a mixture of ketamine (10% at a dose of 50 mg/kg) and xylazine (2% at a dose of 10 mg/kg) were anesthetized and pancreatic tissue and serum of all rats were extracted. Then, UPC3 gene expression levels were measured by Real Time PCR method and MDA serum levels were measured by ELISA method using zellbio kits made in Germany. The sequence of primers of the present research is presented in Table 2. Kolmogorov- Smirnov tests, one-way analysis of variance with Tukey post- hoc test were used to analyze the data (P≤0.05).   Findings The results of one-way analysis test showed that there was a significant difference in UPC3 (P=0.001) and MDA (P=0.001) levels of the studied groups. The results of Tukey post- hoc test showed that UPC3 in diabetic control (P=0.001), training (P=0.008), and eugenol (P=0.02) groups was significantly higher than healthy control group. UPC3 in the training + eugenol group was significantly lower than the diabetic control group (P=0.001) and training group (P=0.001). MDA serum levels in diabetic control groups (P=0.001), training group (P=0.02), and eugenol group (P=0.03) were significantly higher than healthy control group. In addition, UPC3 in training + eugenol group was significantly lower than the control group (P=0.02).   Discussion The results of the present study showed that induction of diabetes had a significant effect on increasing UPC3 gene expression in pancreatic tissue and MDA serum levels. However, aerobic training had no significant effect on pancreatic UPC3 gene expression levels and MDA serum levels in diabetic rats. The findings of some studies show that UPC3 has significant changes due to the consumption of fatty foods, weight gain, as well as under the influence of training and weight loss. Fallahi et al., examined an eight-week period and an acute phase of high-intensity intermittent training (HIIT) on UCP2 and UCP3 in heart tissue of rats and reported an increase in UCP3 expression following the acute phase. They also reported that UCP2 and UCP3 gene expression decreased after a period of HIIT (14). To sum up what was mentioned above, it seems that response to UCPs expression is not related to the type of training; because HIIT was used in the study of Fallahi et al., but it is probably the intensity of the activity that plays a role in the expression of their genes. The researchers said that adaptation to exercise caused the reduction in the expression of UCPs, though; they showed an increase in UCP3 expression after an acute activity. Anyway, increasing or decreasing UCP3 gene expression has been reported inconsistently in various studies (14) and requires further discussion. The results of the present study also showed that eight weeks of eugenol use alone did not have a significant effect on pancreatic tissue UPC3 gene expression levels and MDA serum levels in diabetic rats. However, eight weeks of eugenol consumption along with training had a significant effect on reducing gene expression levels of UPC3 in pancreatic tissue and MDA serum levels in diabetic rats. Moreover, consumption of eugenol with training has a more favorable effect on reducing the expression levels of UPC3 gene in pancreatic tissue and MDA serum levels of diabetic rats than the training alone. In this regard, a significant decrease in the MDA index was observed in the research of Aziz Beigi et al., following incremental resistance training (21). Fetors et al., and Carabolt et al., in their studies also reported a significant decrease in MDA due to the training, the results of which are consistent with the present study (22, 23). Lack of control total calorie intake and calorie consumption of rats along with the measurement of metabolic- cellular parameters is one of the limitations of the present study; therefore, it is suggested that the effect of aerobic exercise with different intensities along with taking nano-eugenolsupplements with different doses on these variables be investigated in the future studies.   Conclusion It seems that aerobic training accompanied by nano-eugenol consumption has more beneficial effects than each of them alone on the improving UPC3 gene expression levels and MDA serum levels in diabetics.   Ethical Considerations Compliance with ethical guidelines This study was approved by the Ethics Committee of the Isfahan (Khorasgan) Branch of Islamic Azad University with ethical code: IR.IAU.KHUISF.REC.1398.178.   Funding Isfahan (Khorasgan) Branch of Islamic Azad University   Authors' contributions Design and conceptualization: Khosro Jalali Dehkordi and Farzaneh Taghian; Methodology and data analysis: Shirin Fotovvat; Supervision and final writing: Khosro Jalali Dehkordi and and Farzaneh Taghian.   Conflicts of interest The authors declared no conflict of interest.  

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