The Effect of Aerobic Training and Nettle Extract on the Gene Expression of IL-8 and Tumor Volume in Mice with Melanoma Cancer

Melanoma cancer is one of the most common types of skin cancer. In this cancer, skin cells grow uncontrollably and divide rapidly. Cytokines play an important role in regulating immune function. Cytokines also play an important role in the onset and proliferation of various cancers. Cytokines, especially interleukin-8, play an important role in tumor growth. Interleukin-8 also plays a role in angiogenesis, tumor volume growth. On the other hand, the use of medicinal plants for the prevention and treatment of diseases is considered by traditional medicine experts. Various studies have shown that nettle extract is involved in several biological and biochemical activities, which have the potential to treat various disorders that affect the skin, gastrointestinal tract, joints, genitourinary system and benign prostatic hyperplasia. In recent years, endurance exercise has been introduced as a safe intervention in preventing and improving the quality of life of people with cancer. However, the therapeutic aspects of exercise training and the mechanisms of effect of this type of exercise on effective indicators and cancerous tumors, especially melanoma, are still debated and less research has been done to identify the effective mechanisms. The aim of this study was to evaluate the effect of nettle extract consumption and aerobic exercise on IL-8 and Tumor volume gene expression in mice with melanoma.

In this experimental study, 20 adult male rats with weighing 300 to 350 g were randomly divided into 4 groups, including groups of: control, exercise, extract and exercise + extract. This research, which is the result of a working group, was approved by the code of ethics No. IR.IAU.M.REC.1399.008 in the Islamic Azad University, Marvdasht Branch. Humidity of 55 5 5% and light cycle were maintained at 12:12 with proper ventilation. Animal feed and water were freely available until the end of the protocol. B16F10 cells were purchased from the Pasteur Institute of Iran. These cells were selected because the cell type was the same as the studied mouse species. The cells were cultured in M199 medium and when the cell density reached 80%, they were prepared for injection into mice.Exercise training was performed on a treadmill for six weeks and 5 sessions per week. Mice were trained for 10 to 15 minutes at a speed of 10 meters per minute for 5 days in order to get acquainted with the treadmill for a week. From the second week, the overload phase was performed for three weeks until the end of the fourth week. In the next steps and every day of training, 3 minutes of activity time and one meter per minute were added to the treadmill speed. At the end of the fourth week, the speed of the treadmill reached 28 meters per minute for 60 minutes of activity.One week after induction of melanoma cancer and through implantation of the tumor under the skin. To prepare the extract of nettle, some stems and leaves of nettle were collected and washed in small pieces, then dried in the open air and powdered. Then aqueous extract of nettle was prepared. the experimental group consumed 30 mg / kg / day of nettle ethanol extract orally for 8 weeks. Sampling was performed 48 hours after the last session of endurance activity. RT PCR was used to measure the expression of IL-8gene . Shapiro-Wilk test was used to determine the normality of data distribution and Levin test was used to examine the homogeneity of variance. Also, to investigate the significant changes in each of the research variables, one-way analysis of variance was used between different groups and if a statistically significant difference was observed, Tukey post hoc test in ANOVA program was used to determine the location of intergroup differences.

The results of the present study showed that consumption of nettle extract and aerobic exercise significantly reduced the expression of interleukin 8 in the experimental groups compared with the control group (p = 0.125, p = 0.278, = 0.174, respectively. p). IL-8 gene expression decreased in experimental groups compared to control group; But it did not reach a significant level. These results showed that physical exercise reduced the expression of IL8 gene and also these values were reduced in the extract group and the extract and exercise (combined) group, but the relevant values were not significant.The results also showed that tumor volume was significantly reduced among the experimental groups compared to the control group (p = 0.021, p = 0.136, p = 0.047, respectively). The results also showed that tumor volume in the experimental groups was significantly reduced compared to the control group (p = 0.003). The results of this study showed that exercise and exercise and extract (combined) could cause a significant reduction in tumor volume, but the reduction in the extract group was not significant compared to the control group.

The data of the present study show that endurance training combined with nettle extract has an effective role in reducing the expression of cytokine interleukin 8 in mice with melanoma and since the reduction of this cytokine is associated with a decrease in tumor volume. Overall, it can be concluded that endurance training can modulate the levels of angiogenic cytokines within the tumor and also regulate inflammatory cytokines. Also in tumor tissue, cytokines are produced by tumor cells. In mice, treatment with nettle extract is likely to be enhanced due to increased antioxidant mechanisms. The activity of catalase (CAT) and superoxide dismutase (SOD) enzymes and glutathione content (GSH) were also increased. One of the prominent symptoms of cancer is resistance to apoptosis, which indicates that the induction of apoptosis in cancer cells is an important anti-cancer mechanism. In addition, patoltin inhibits proliferation by an apoptotic activity in several tumor cell lines. Its role in inhibiting the proliferation of cancer cell lines occurs by activating apoptotic pathways. Nettle extract contains other compounds such as gallic acid, chlorogenic acid and homovanilic acid, all of which are associated with anti-cancer properties. These activities may also be attributed to the flavonoid content of nettle. According to the results of this study and some similar studies, it can be concluded that physical activity and consumption of nettle extract can play an effective role in controlling the progression of melanoma cancer, reducing tumor volume, prevention and even treatment of melanoma cancer.