1. Introduction
Doxorubicin (Dox) is a drug from the family of anthracyclines, which is used in the treatment of a wide range of malignancies such as lymphoma and leukemia [1, 2]. In addition to the anti-tumor effects of this drug, side effects such as the toxicity of healthy tissues including heart tissue, have caused a serious problem in the use of this drug for the treatment of cancers [3]. There are several methods to reduce the side effects caused by Dox toxicity, such as limiting the cumulative dose of the drug, reducing its consumption, and performing continuous aerobic activity [3, 9]. Moderate intensity physical activity, as one of the appropriate protective strategies, can protect the heart against Dox-induced cardiotoxicity [10]. Metabolic, muscular and cardiovascular adaptations in healthy and affected groups depend on the intensity of physical activity. The beneficial effects of high intensity interval training (HIIT) are much greater and more acceptable than continuous aerobic training in reducing inflammatory and traditional risk markers, despite the short time and reduced amount of activity [12, 13]. In addition to physical activity, medicinal plants with different biological properties have protective effects against the toxicity caused by chemical substances due to having no side effects [18]. Crocin, which is one of the effective components in the saffron plant, has strong antioxidant and anti-inflammatory effects [19]. This study aims to investigate the combined effect of HIIT and crocin supplementation on serum markers of cardiac damage including lactate dehydrogenase (LDH) and creatine kinase (CK-MB) isoenzyme.
Methods
In this study, 32 male Wistar rats with an average weight of 200-220 g were prepared and kept in suitable conditions. Then, they were randomly divided into four groups: healthy control (saline), doxorubicin (patient control), doxorubicin + HIIT, and doxorubicin + HIIT + crocin. The last two groups performed the HIIT for 8 weeks, 5 days per week in the form of 2-minute intervals of high and low-intensity running on the treadmill. Crocin was prepared as a ready-made powder in 1 and 5 g vials with a purity of 98% (Sigma, USA). The last group received 10 mg/kg body weight of crocin dissolved in 10 mg/kg normal saline orally (by gavage) on training days. Doxorubicin was purchased from the ABO company, Belgium. Then, seven doses were diluted with normal saline to prepare the desired dose and was injected subcutaneously seven times at the end of each week. LDH and CK-MB levels were measured using special laboratory kits according to the instructions of the manufacturer and by spectrophotometric method. To determine the level of CK activity, CK-MB isoform kit (Pars Azmoun, Tehran, Iran) was used. The standard method proposed by the German Society of Clinical Chemistry (DGKC) was used to measure LDH activity [26]. The obtained data were analyzed using appropriate statistical tests. The significance level was set at 0.05.
Results
The data related to the mean and standard deviation of the study variables for each group are presented in Table 1.


The results showed that the lowest levels of CK-MB and LDH enzymes were in the healthy control group and the highest levels were in the doxorubicin group (Table 1). Data analysis showed that injection of doxorubicin caused a significant increase in serum levels of LDH and CK-MB compared to the healthy control group (P=0.001) (Table 2).


There was no significant difference between the effects of HIIT alone and HIIT plus crocin consumption on the serum levels of LDH (P=0.087) and CK-MB (P=0.877) in rats received doxorubicin (Figures 1 and 2). Discussion
The results showed that the Dox injection led to a significant increase in LDH and CK-MB levels in rats, but HIIT alone and in combination with crocin consumption reduced the changes in serum levels of LDH and CK-MB induced by doxorubicin. There was no significant difference between the effects of HIIT alone and HIIT plus crocin consumption on the serum levels of LDH and CK-MB in rats received doxorubicin.

Ethical Considerations
Compliance with ethical guidelines
This study was approved by the ethics committee of Lorestan University of Medical Sciences (Code: IR.LUMS.REC.1399.221).

Funding
This study was extracted from the master thesis of first author and funded by Lorestan University of Medical Sciences.

Authors' contributions
All authors equally contributed to preparing this article.

Conflicts of interest
The authors declared no conflict of interest.

Acknowledgements
The authors would like to thank the Vice-Chancellor for research of Lorestan University of Medical Sciences for their support.


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