Document Type : Original Article
Authors
1 Ph.D Student, Department of exercise physiology, Karaj branch, Islamic Azad University, Karaj, Iran
2 Associate Professor, Department of exercise physiology, Karaj branch, Islamic Azad University, Karaj, Iran
3 Assistant Professor, Department of exercise physiology, Karaj branch, Islamic Azad University, Karaj, Iran
Abstract
Extended Abstract
Background and Aim: Non-alcoholic fatty liver disease (NAFLD) is an acquired metabolic disorder characterized by the deposition of triglycerides in the liver, caused by factors other than alcohol consumption(1). Currently, the treatment of NAFLD primarily involves physical activity(9) and dietary supplements to reduce fat mass(16). Mitochondrial abnormalities, such as dysfunction in the electron transport chain and reduced beta-oxidation of fatty acids, contribute to fat accumulation in liver cells, leading to cellular damage and progression to non-alcoholic steatohepatitis (NASH)(2). Exercise not only reduces liver inflammation but also enhances mitochondrial biogenesis, improving morphological dynamics and regeneration rates through biogenesis and mitophagy(10). Additionally, insect-based supplements, such as mealworms, have demonstrated strong immunostimulatory, anticancer, antidiabetic, and antioxidant properties(17). This supplement, rich in protein, has been reported to exhibit anti-obesity effects by activating PGC-1α. Furthermore, it has been shown to effectively reduce weight and fat mass by influencing the expression of genes involved in fat metabolism(18). in this study, the simultaneous effect of mealworm protein supplementation with aerobic exercise on the expression of genes involved in mitochondrial biogenesis, including peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), uncoupling protein 1 (UCP1), mitofusin 1 (Mfn1), and dynamin-related protein 1 (Drp1), in the soleus muscle tissue of rats with NAFLD.
Materials and Methods: In this experimental study, 25 male Wistar rats were divided into five equal groups including: 1- healthy, 2- patient, 3- patient+ supplement, 4- patient+ exercise, 5- patient+ supplement + exercise. The sick groups developed NAFLD with high fat and cholesterol diet. Interval endurance exercise was performed for eight weeks, including running on a treadmill for 30 minutes, five days a week with gradual overload. mealworm supplement gavage with a dose of 20 mg/kg body weight was performed on the same days as exercise. Tissue sampling of the soleus muscle was performed 48 hours after the last exercise and supplement session. The animals were anesthetized with intraperitoneal injections of ketamine and xylazine, and the soleus muscle tissue was immediately collected. Each tissue sample was placed in a microtube containing liquid nitrogen and stored at -80°C until gene expression analysis. RNA extraction was performed using the RNX-Plus method, and the quality and quantity of RNA were assessed via spectrophotometry and agarose gel electrophoresis. The sequences of mRNAs corresponding to the genes PGC-1α, UCP1, Mfn1, and Drp1 were extracted using the NCBI database. Primers were designed using AllellD software and evaluated for specificity using BLAST. The obtained data were analyzed using independent t-tests and two-way ANOVA at a significance level of p ≤ 0.05.
Results: The independent t-test results showed that in the diseased group, the expression of PGC-1α, UCP1, and Mfn1 genes significantly decreased compared to the healthy group, while the expression of Drp1 significantly increased (p = 0.0001). The two-way ANOVA results indicated that aerobic exercise significantly increased the expression of PGC-1α (p = 0.0001), UCP1 (p = 0.0001), and Mfn1 (p = 0.0001), while significantly decreasing the expression of Drp1 (p = 0.0001) in the soleus muscle tissue of rats with NAFLD. Additionally, statistical findings revealed that tenebrio molitor protein supplementation significantly increased the expression of UCP1 (p = 0.0001), PGC-1α (p = 0.0001), and Mfn1 (p = 0.0001), while significantly decreasing the expression of Drp1 (p = 0.0001) in the soleus muscle tissue of rats with NAFLD. However, the two-way ANOVA results showed that the combination of aerobic exercise and menebrio tolitor protein supplementation did not have a significant effect on the expression of UCP1 (p = 0.537), PGC-1α (p = 0.361), or Drp1 (p = 0.881) in the soleus muscle tissue of rats with NAFLD. Nevertheless, the simultaneous intervention of aerobic exercise and mealworm protein supplementation significantly increased the expression of Mfn1 (p = 0.0001) in the soleus muscle tissue of rats with NAFLD.
Table 1. Results of two-way ANOVA comparing relative gene expression changes in the study groups
*Significant effect of aerobic exercise on gene expression. **Significant effect of supplement on gene expression. ***Significant interaction effect (p ≤ 0.05).
Conclusion: Aerobic exercise and mealworm protein supplementation each independently led to a significant increase in the expression of PGC-1α, UCP1, and Mfn1 genes, as well as a significant decrease in the expression of Drp1 in the soleus muscle of rats with NAFLD. It appears that moderate-intensity interval aerobic exercise and mealworm extract supplementation, individually, can positively influence the levels of key genes involved in mitochondrial biogenesis and fat metabolism, potentially aiding in the treatment of NAFLD. Further studies are recommended to explore the interactive effects of exercise and mealworm supplementation on mitochondrial capacity and fat metabolism, particularly in samples with obesity or type 2 diabetes associated with NAFLD.
Keywords: Aerobic exercise, mealworm Larvae, mitochondrial biogenesis, Fatty Liver
Hical Considerations: All stages of animal care and euthanasia were conducted in accordance with the Helsinki Declaration (1964).
Funding: Ethical approval was obtained from the Ethics Committee of Islamic Azad University, Karaj Branch (Code: IR.IAU.K.REC.1403.029).
Conflict of Interest: No conflicts of interest declared.
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