The effect of high-intensity interval training (HIIT) on the expression of proteins involved in autophagy, apoptosis, and atrophy pathways in the myocardium of male rats with type 2 diabetes

doi:10.21203/rs.3.rs-2105962/v1

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Research Article

The effect of high-intensity interval training (HIIT) on the expression of proteins involved in autophagy, apoptosis, and atrophy pathways in the myocardium of male rats with type 2 diabetes

https://doi.org/10.21203/rs.3.rs-2105962/v1

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Introduction

Autophagy, apoptosis, and atrophy pathways are responsible for controlling cardiac cell homeostasis. This study aims to investigate the effect of high-intensity interval training (HIIT) on the expression of proteins involved in autophagy, apoptosis, and atrophy pathways in the myocardium of type 2 diabetic rats.

Methods

In this experimental study, 18 male Sprague-Dawley rats (2 months old, weighing 280 ± 20 g) were selected. 12 rats were made diabetic through intraperitoneal injection of STZ and nicotinamide solutions. These rats were randomly divided into 2 groups, HIIT + diabetes (HIIT + D), and diabetic (6 rats in each group). A control group (6 rats) was also considered. The HIIT + D group did HIIT protocol 4 days a week for 8 weeks. To analyze the data, one-way ANOVA and Tukey's post hoc tests were used.

Results

It was shown that HIIT leads to a significant increase in AKT and mTOR protein content between the HIIT + D and diabetic groups in the left ventricle. But there is no significant change in the content of AMPK proteins. The content of FOXO3a, Beclin1, P53, myostatin, and SMAD2/3 proteins showed a significant decrease in the HIIT + D group. The protein content of caspase-3 in the activated form did not show a significant change, but there was a significant decrease in the content of the initial form (Pro).

Conclusion

HIIT along with increasing regulation of AKT and mTOR proteins can disable autophagy, apoptosis, and atrophy pathways. Therefore, the process of apoptosis and atrophy of heart cells is reduced in type 2 diabetic rats.

high-intensity interval training

myocardium

autophagy

apoptosis

atrophy

type 2 diabetes

Diabetes is a major public health problem. In 2010, there were about 285 million people with diabetes and it is estimated that this number will increase to about 700 million people in 2040 (1). Diabetes is a background for the disorder in the myocardium of diabetic people (2), which is the main cause of death in diabetic patients (3). The myocardial disorder is characterized by disturbances in cardiac hypertrophy, autophagy, and apoptosis pathways (4). In people with myocardial disorders, autophagy and apoptosis pathways are increased as cell death processes (3, 5). While in a healthy organism, cell proliferation and cell death are in a state of balance (6).

In the myocardium, AKT and mTOR proteins play a vital role in cell survival and growth in physiological and pathological conditions such as exercise, heart diseases, and diabetes. AKT and mTOR proteins can cause protein synthesis, disable autophagy, and disable apoptosis (7–9). The mTOR protein is activated when nutrients are available for cell growth, while the AMPK protein is stimulated under conditions of cell energy deficiency (10–13). Since the AMPK pathway is an inhibitor of the mTOR pathway (14), the mTOR pathway as the main anabolic pathway in diabetes can be impaired and autophagy can increase in diabetes (15).

Autophagy is an intracellular catabolic process that produces energy through the breakdown of damaged proteins and organelles (16–18). One of the important pathways of autophagy in the myocardium of diabetic subjects is the FOXO3a/Beclin-1 pathway (5). The Forkhead Box (FOXO) family of proteins is known to regulate the expression of autophagy genes (19). FOXO3a enhances the expression of autophagy-related genes and maintains cardiomyocyte size by inhibiting cardiomyocyte growth and stimulating autophagy (20, 21). FOXO3a upregulates Beclin-1 in response to cellular stress because it is a master regulator of autophagy in mammalian cells (22–24). At the initiation stage of autophagy, Beclin-1 forms a double-membrane structure to form the autophagosome (25, 26). Decreased Beclin-1 levels are thought to inhibit autophagy and prevent the turnover of damaged mitochondria, leading to the production of reactive oxygen species (ROS) (27).

The apoptosis pathway is another type of programmed death through ROS, which through intrinsic and extrinsic signals leads to the elimination of damaged cells and inhibits the proliferation of abnormal cells (6, 28, 29). P53 protein plays an important role in the regulation of apoptosis and promotes the apoptotic pathway (29, 30). Caspase 3 is another effector of apoptosis, which in turn is responsible for breaking down many important proteins. Caspase 3 also activates caspases 6, 7, and 9 to promote the apoptotic pathway (31, 32).

Since cardiac hypertrophy plays an important role in cardiac regeneration, understanding its inhibitory factors is very important. Myostatin is a member of the large TGF-β family, which is a negative regulator of muscle differentiation and growth (33). Myostatin is expressed in skeletal muscle and heart (34, 35). The myostatin-SMAD pathway plays an essential role in controlling muscle size. Factors such as immobility usually increase myostatin mRNA (36). Myostatin is activated only when it binds to the TGFβ type II receptor, then it phosphorylates Smad2/3. Phosphorylated Smad2/3 enters the nucleus together with Smad4 and regulates negatively genes controlling cardiac growth (37).

Today, HIIT is considered one of the non-invasive methods for the harmful effects of diabetes. This type of exercise is a combination of high-intensity periods with moderate to low-intensity active or passive recovery periods. Studies have shown that with high-intensity exercise (85–90% of VO2max) compared to moderate-intensity exercise (65–70% of VO2max), twice the beneficial cardiac effects can be obtained (38). HIIT has been reported to lead to improved cardiac function, cardiac remodeling, and aerobic capacity as an adaptive response in humans and rodents (39). HIIT-induced cardiac adaptations have been characterized by improved circulatory function, increased antioxidant capacity, and mitochondrial respiration. However, the cellular and molecular adaptations in the myocardium are unknown (40), and the physiological effects of HIIT on myocardial function are poorly understood. Because very little research has been done on the pathways of autophagy, apoptosis, and myocardial atrophy in diabetic people.

Since the function of these proteins after HIIT is unclear, identifying the function of proteins and cellular pathways in autophagy, apoptosis and myocardial hypertrophy can be useful. Therefore, this research aims to investigate the effect of HIIT on the content of autophagy (FOXO3a and Beclin1), apoptosis (P53 and caspase-3), and atrophy (myostatin and SMAD2/3) proteins through upstream regulatory proteins (Akt, AMPK, and mTOR) in the myocardium of male rats with diabetes.

Sample and type of research

The current research is an experimental study. In this research, 18 male Sprague Dawley rats (2 months old, weighing 280±20 grams) were selected. The rats were kept in the animal laboratory of Shiraz University of Medical Sciences with a light-dark cycle of 12-12 hours and an air temperature of 22±2°C with a humidity of 40-50%. Animal food and water were provided freely and standardly to the rats. Ethical principles (ethical code IR.SUMS.REC.1396.S1062) were considered according to the principles of working with laboratory animals approved by Shiraz University of Medical Sciences.

Diabetes induction method

12 rats were randomly selected to develop type 2 diabetes, and in the first stage, nicotinamide solution was injected with a dose of 110 mg per kilogram of body weight. Then, 15 minutes later, streptozotocin (STZ) solution (dissolved in 0.1 M citrate buffer with pH = 4.5) was injected intraperitoneally and it was done only once with a dose of 60 mg per kilogram of body weight (41). To ensure that the rats became diabetic, their blood sugar was measured 72 hours after the injection using a blood sugar measuring device. Blood samples were taken from the tail vein of mice, and blood sugar between 126 and 260 mg/dL was considered an indicator of type 2 diabetes (42).

HIIT protocol

Diabetic rats (12 heads) were randomly divided into the HIIT+D (6 rats) and the diabetic groups (6 rats). A control group (6 heads) was also considered. The HIIT+D groups ran on the treadmill at a speed of 5 to 10 meters per minute for a week to familiarize themselves with the treadmill. The program of the HIIT+D group was for 8 weeks and 4 sessions per week. The rats in the HIIT+D group warmed up at a speed of about 10 to 12 meters per minute. Then the exercise program included 5 periods of 4 minutes with an intensity equal to 85 to 95% of the maximum speed and active rest periods of 3 minutes with an intensity equal to 50 to 60% of the maximum speed. At the end of each session, the rats cooled down at a speed of about 10 to 12 meters per minute. The total duration of running in each session was 44 minutes. The tilt of the turntable was 0 degrees and did not change in 8 weeks (43).

Speed measurement test

The test started with a speed of 5 meters per minute and the speed increased by 5 meters per minute every 3 minutes. When the rats reached the end of the treadmill (sticking the rats to the end of the treadmill), the speed of the rats was considered the maximum speed (44).

Tissue removal method

After 24 hours after the last training session, rats were anesthetized by intraperitoneal injection of ketamine (30-50 mg/kg body weight) and xylazine (3-5 mg/kg body weight). Then, the tissue of the left ventricle of the heart was removed from the animal's body and washed in physiological serum. Then it was immediately frozen using nitrogen and frozen at -80 for further measurements.

Western blot laboratory methods

The variables of the research were measured using the western-blot method. To extract the proteins of left heart ventricular tissue from RIPA buffer containing 0.05 mM Tris buffer (pH 8), 150 mM sodium chloride, 0.01% EGTA, 1% sodium dodecyl sulfate (SDS) plus 0.1% antiprotease. (sigma) cocktail was used. 100 mg of tissue was homogenized in a 500 μL buffer containing antiprotease by a homogenizer. Then it was kept at 4°C for half an hour and centrifuged in a refrigerated centrifuge (bo, sw14rfroil) at 12000 rpm and 4°C for 10 minutes. Then the supernatant was collected and its protein concentration was measured with a protein determination kit (Bio-Rad) (it was measured at a wavelength of 595 nm). Finally, it was kept at 20 degrees below zero, then the obtained homogenate was mixed in a ratio of 1:1 with the loading buffer sample (50 mM tris-hydrogen chloride, 2% sodium dodecyl sulfate, 10% glycerol, 5% beta-mercaptoethanol and 0.005 percentage of aqueous bromophenol) was mixed. Next, the samples were boiled for 5 minutes to completely denature all the proteins. Proteins were separated using SDS-Polyacrylamide gel electrophoresis. Then they were transferred to a nitrocellulose membrane. The membrane was blocked for 1 hour in 5% BSA in Tris-Buffered Saline and 0.1% (Tween 20 TBST) and incubated in primary antibody (500:1). Incubation in secondary antibody was performed the next day for 1 hour at room temperature in 4% TBST. Proteins were quantified by a chemiluminescence (ECL) reaction and by densitometry analysis with Image J software (version 1.8.0.112) (45).

Statistical analysis

First, the Kolmogorov-Smirnov test was used to determine the normality of the research data. Due to the normality of data distribution, one-way ANOVA and Tukey's post hoc tests were used. Data analysis was done using SPSS version 23 software. A significance level of P≤0.05 is considered.

The results of our study showed that the weight (gr) of rats in the diabetes group increased significantly in the 8th week compared to the first week (p < 0.002). In addition, rats’ weight (g) in the control group was significantly increased after 8 weeks (p < 0.009). Additionally, the HIIT + D group’s body weights increased after 8 weeks in comparison to the first week (p < 0.001). Blood glucose of control groups after 8 weeks wasn’t changed significantly (p < 0.72). Furthermore, blood glucose (mg/dl) in the diabetes group was significantly higher in the 8th week than in the first week (p < 0.006). However, there were no significant changes in blood glucose in HIIT + D groups after 8 weeks of training compared to the first week (p < 0.12) (Table 1).

Table 1

Paired t-test for weight (g) and blood glucose (mg/dl)
variable	Group	Mean	SD	t	P-value
Weight (g)	Control (1th week)	284.30	4.91	4.82	0.009
	Control (8th week)	299.10	3.20	4.82	0.009
	Diabetes (1th week)	259.20	11.07	7.35	0.002 ^‡
	Diabetes (8th week)	296.20	12.19	7.35	0.002 ^‡
	HIIT + D (1th week)	251.80	9.70	8.84	0.001^#
	HIIT + D (8th week)	263.20	11.69	8.84	0.001^#
Blood glucose (mg/dl)	Control (1th week)	82.60	2.07	0.37	0.72
	Control (8th week)	83.00	2.54	0.37	0.72
	Diabetes (1th week)	222.00	33.25	5.31	0.006 ^†
	Diabetes (8th week)	292.00	15.34	5.31	0.006 ^†
	HIIT + D (1th week)	236.20	39.95	1.95	0.12 ^ns
	HIIT + D (8th week)	251.00	30.04	1.95	0.12 ^ns
Data are expressed as mean ± SD. HIIT: High-Intensity Interval Training. ns: non-significant
^‡ Rats’ weight (g) in the control group was significantly increased after 8 weeks (p < 0.009).
^‡ Rats’ weight (g) in the diabetes group was significantly increased after 8 weeks (p < 0.002).
^# HIIT + D group’s weight was increased after 8 weeks (p < 0.001).
^† Blood glucose (mg/dl) in the diabetes group was significantly higher in the 8th week than in the first week (p < 0.006).
Blood glucose of control groups after 8 weeks wasn’t changed significantly (p < 0.72).
Blood glucose of HIIT + D groups after 8 weeks wasn’t changed significantly (p < 0.12).

The content of AKT, mTOR and AMPK proteins

The data showed that after performing HIIT, there is a significant increase in AKT and mTOR protein content (P = 0.0001); Tukey's post hoc test showed that this change is significant between HIIT + D groups compared to diabetic (P = 0.0001), HIIT + D compared to control (P = 0.0001) and control compared to diabetic (P = 0.0001) (Fig. 1, A and B ).

Performing HIIT led to a significant change in the content of AMPK proteins (P = 0.0001). Tukey's post hoc test showed that this increase was significant between HIIT + D groups compared to the control (P = 0.0001) and diabetic compared to the control (P = 0.0001). But no significant change was observed between HIIT + D groups compared to the diabetic (P = 0.299) (Fig. 1, A, and B ).

The content of proteins involved in the autophagy pathway, FOXO3a, and Beclin1

The content of FOXO3a and Beclin1 proteins showed a significant decrease after HIIT (P = 0.0001). Tukey's post hoc test showed that this change was significant between the HIIT + D group compared to diabetics (P = 0.0001) and the control compared to diabetics (P = 0.0001). A significant level was reported for FOXO3a and Beclin1 proteins in HIIT + D group compared to the control (P = 0.012) and diabetics (P = 0.0001), respectively (Fig. 2, A, and B).

The content of proteins involved in the apoptotic pathway of P53 and pro and active caspase-3

P53 protein content showed a significant decrease (P = 0.0001). Tukey's post hoc test showed that this change was significant between diabetic groups compared to HIIT + D (P = 0.0001), HIIT + D compared to control (P = 0.0001), and also diabetic compared to control (P = 0.0001) (Fig. 3, A, and B ).

The protein content of the initial caspase-3 also showed a significant decrease (P = 0.001). Tukey's post hoc test showed that this change was significant between HIIT + D groups compared to diabetics (P = 0.004) and control compared to diabetics (P = 0.001). No significant change was observed between HIIT + D groups compared to control ones (P = 0.718) (Fig. 3, A, and B ).

The protein content of caspase-3 in the activated form showed a significant change (P = 0.0001). Tukey's post hoc test showed that this change is significant between HIIT + D groups compared to control (P = 0.0001) and also control compared to diabetic (P = 0.001). No significant change was observed between HIIT + D groups compared to diabetics (P = 0.471) (Fig. 3, A, and B ).

The content of proteins related to cardiac atrophy myostatin and SMAD2/3

Myostatin protein content showed a significant decrease (P = 0.0001). Tukey's post hoc test showed that this change was significant between the groups of HIIT + D compared to diabetic (P = 0.0001), HIIT + D compared to control (P = 0.009), and also control compared to diabetic (P = 0.0001) (Fig. 4, A and B ).

SMAD2.3 protein content showed a significant decrease (P = 0.0001); Tukey's post hoc test showed that this change was significant between HIIT + D groups compared to diabetics (P = 0.0001) and control compared to diabetics (P = 0.0001). No significant change was observed between HIIT + D groups compared to control ones (P = 0.652) (Fig. 4, A and B ).

The results of the present study showed that the implementation of HIIT leads to a significant increase in the protein content of AKT and mTOR between the diabetic and HIIT + D groups in the myocardium. While there is no significant change in the content of AMPK proteins. On the other hand, the content of FOXO3a, Beclin1, P53, myostatin, and SMAD2/3 proteins showed a significant decrease in the HIIT + D group. While the active caspase-3 protein content did not show a significant change but the content of the initial form of caspase 3 decreased significantly.

It seems that performing HIIT in diabetic rats can deactivate AMPK protein by activating AKT and mTOR proteins. This process plays a critical role in cell survival and growth under physiological conditions and induces protein synthesis, inactivation of autophagy, and inactivation of apoptosis (7–9, 46). The results of Sturgeon et al.'s research are in line with the results of the current research in changing the content of AMPK protein, but they are contradictory in changing the content of AKT and mTOR proteins. The results of the study by Sturgeon et al. (2015) showed that moderate intensity exercise does not lead to significant changes in the content of AKT, mTOR, and AMPK proteins in the myocardium. While exercise improves cardiac function and exercise capacity, causing physiological hypertrophy in the heart (47). One of the important reasons for this contradiction is the intensity and type of exercise. In the present research, the exercise is high-intensity and intermittent, but in the research of sturgeon et al., the exercise is moderate-intensity and continuous. It seems that HIIT can be similar to resistance exercises that play an important role in protein synthesis and myocardial hypertrophy (11).

On the other hand, reducing autophagy factors such as FOXO3a and Beclin1 through HIIT can be beneficial for people with diabetes because diabetes is associated with myocardial dysfunction, which can be caused by autophagy dysfunction (48). The results of Holloway et al.'s research on the change of FOXO3a protein content are contradictory to the results of the current research because it was reported that HIIT exercises do not affect the content of FOXO3a protein in the myocardium of rats suffering from heart failure (49). One of the reasons for the contradictions is the type of disease because, in the present study, the samples are suffering from diabetes, but in Holloway et al.'s research, the samples are suffering from heart failure. On the other hand, the research results of Zeng et al. (2020) are in line with the results of the current research on FOXO3a protein content. But it is contradictory in Beclin1 protein content because Beclin1 protein content increases after resistance, aerobic, combined (aerobic + resistance) exercises in the skeletal muscle of old rats (50). One of the important reasons can be the place of protein content measurement because protein measurement was done in heart tissue in the present study and skeletal muscle tissue in Zeng et al.'s research. In addition, in the present study, the samples were diabetic rats, and in the research of Zeng et al., the samples were reported to be old rats suffering from sarcopenia. Also, Marfe et al. (2012) showed that FOXO3a transcription decreases in heart muscle tissue after a HIIT session in rats (51). While Li et al. (2018) found that HIIT is associated with an increase in Beclin-1 protein content (52). Therefore, the results of Li et al.'s research are contradictory to the results of the present research.

Therefore, it seems that diabetic rats are prone to impaired autophagy in the myocardium due to disruption of the PI3K/AKT/mTORC1 pathway (53). Because other studies have confirmed that rats lacking the insulin receptor (IRS) promote autophagy in the heart (54). Hence, it can be concluded that HIIT can decrease the autophagic proteins FOXO3a and Beclin-1 in the myocardium by activating the PI3K/AKT/mTORC1 pathway. In support of this, other studies have reported that decreased autophagy in the heart of diabetic patients is due to increased mTORC1 pathway activity due to the inactivation of AMPK (55). It seems that the mTORC1 pathway can inhibit or activate autophagy by regulating ULK1, ATG13, and FIP200 proteins (56). Therefore, probably HIIT can prevent myocardial autophagy disorder and maintain cell stability (57–59). Therefore, exercise training is an important non-pharmacological approach that can be used to increase the quality of life and reduce pathological symptoms (60).

Excessive activation of apoptosis is one of the damages of diabetes in the myocardium because recent studies show an increase in apoptosis in the heart of diabetic patients (27, 61, 62). In the present study, a significant decrease in p53 protein content occurred after HIIT, which is in contrast to Sadeghi et al.'s (2020) study, as they reported an increase in p53 protein content after exercise (63). This contrast seems to be due to a lack of adaptation to exercise because after a non-exercise period the increase in p53 tended to decrease in the long-term exercise group (63). Also, other studies have shown that exercise training can lead to the inactivation of mTORC1, and activation of autophagy and apoptosis by activating AMPK and P53 (64). While the current research has shown an increase in mTORC1 and a decrease in p53 after HIIT. HIIT seems to decrease P53 due to unchanged AMPK and increased mTORC1. Therefore, factors such as adaptation, recovery, duration, and intensity of training can affect p53.

In the present study, the content of initial caspase-3 protein (Pro) was significantly decreased in the diabetic and healthy exercise groups compared to the diabetic group. But on the other hand, the active caspase-3 protein content increased in the diabetic exercise group compared to the healthy and diabetic groups. One reason for the increase in active caspase-3 is the lack of adaptation in rats to HIIT exercise at 8 weeks. These results are in line with the study of vahid et al. (2022) as they showed that HIIT resulted in a significant increase in Caspase-3 (65). Because HIIT causes an inflammatory response by increasing oxidative stress and the mitochondrial membrane becomes unstable, then cytochrome C can be released and facilitate the process of apoptosis (66). In addition, the underlying mechanisms for increased apoptosis include increased ROS, increased inflammatory cytokines, activation of caspases, Fas receptor-dependent and mitochondria-dependent apoptosis, ER stress, increased activation of the TGF-β signaling pathway, and IGF-1 resistance (67–70). Since lack of adaptation to exercise can trigger apoptosis, it seems that 8 weeks is not the right time to adapt to HIIT. However, adapting to sports activities can lead to the modulation of oxidative stress by increasing the antioxidant capacity, and reduce the factors associated with the activation of the apoptosis pathway (66).

The results of lenk et al.'s research are in line with the results of the present research. In the research of Lenk et al. (2009) (2012), the expression of myostatin protein increases in rats suffering from chronic heart failure (CHF), but after endurance training, a significant decrease in the expression of myostatin protein is seen in the myocardium. These investigators stated that downregulation of myostatin has anti-catabolic effects on CHF and halts myocardial destruction in chronic heart failure (71, 72) because the myostatin/SMAD pathway leads to reduced activity of the mTOR pathway (73). However, in both previous studies, despite the difference in the type of exercise (HIIT and endurance), myostatin expression is decreased. Therefore, it seems that exercise training can prevent muscle loss (74) and lead to an increase in the activity of the mTOR pathway, which was shown in the present study. On the other hand, in the research of Kabak et al. (2018), it was reported that the level of myostatin protein increases immediately after HIIT training; But 3 hours after training, it shows a tendency to return to the basic state (75). It seems that it is normal to increase myostatin (a negative muscle growth factor) during exercise due to muscle damage, and it makes sense for myostatin to decrease during recovery for myocardial growth. Therefore, in the present study, the reduction of myostatin and the increase of the mTOR pathway after HIIT seems reasonable.

The results of Launay et al.'s research are in line with the results of the present research because they showed that 8 weeks of HIIT training (95% intensity) leads to a decrease in SMAD2/3 proteins in the left ventricle through the mTORC1 pathway (76). It seems that exercise training can activate the signaling cascade of insulin and growth factors and then lead to the activation of AKT and mTOR proteins. The activation of these proteins can affect the reduction of myostatin and SMAD protein and lead to cardiac hypertrophy (77). In another study, the mean values of SMAD3 gene expression were significantly reduced by performing HIIT in the left ventricle of rats with type 2 diabetes. These researchers stated that HIIT could improve diabetic cardiomyopathy by affecting the reduction of SMAD3 gene expression (78). Based on the cellular pathway of myostatin and SMAD, the increase of myostatin protein leads to the activation of SMAD proteins, which play an important role in atrophy and heart defects (79). Therefore, it seems that exercise training with appropriate intensity, duration, and recovery leads to modulation of ROS, intracellular calcium concentration, cellular energy status, autophagy, apoptosis, and adaptation (80). Hence, it appears that HIIT can prevent excessive myocardial autophagy, apoptosis, and atrophy in people with type 2 diabetes.

The results of the present research showed that HIIT leads to an increase in AKT and mTOR proteins, while it does not affect AMPK protein. It seems that the regulation of upstream proteins leads to a decrease in the expression of autophagy (FOXO3a and Beclin1), apoptosis (P53, initial and active caspase-3), and atrophy (myostatin and SMAD2/3) proteins in the cardiac muscle of diabetic patients. Then, this process prevents excessive autophagy, apoptosis, and atrophy. Therefore, HIIT can be recommended as a non-pharmacological approach for people with diabetes to prevent myocardial disorders.

high-intensity interval training (HIIT), HIIT+diabetes (HIIT+D), Forkhead Box (FOXO), reactive oxygen species (ROS), streptozotocin (STZ), chronic heart failure (CHF).

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No competing interests reported.

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The effect of high-intensity interval training (HIIT) on the expression of proteins involved in autophagy, apoptosis, and atrophy pathways in the myocardium of male rats with type 2 diabetes

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