The effects of 12 weeks of swimming training and royal jelly consumption on the expression of JNK and AP-1 genes of lung tissue in mice with lung cancer

Rasoulizadeh, Hossein; Moghadasi, Mehrzad; Hosseini, Seyed Ali Hosseini

doi:10.48308/joeppa.2025.240386.1372

The effects of 12 weeks of swimming training and royal jelly consumption on the expression of JNK and AP-1 genes of lung tissue in mice with lung cancer

Document Type : Original Article

Authors

¹ Department of sport sciences, Shi.C., Islamic Azad University, Shiraz, Iran

² Department of sport sciences, Marv.C., Islamic Azad University, Marvdasht, Iran

10.48308/joeppa.2025.240386.1372

Abstract

Background and Purpose: Lung cancer is one of the most important causes of global mortality and oxidative stress and inflammatory factors play an important role in its occurrence and development. Probably regular exercise along with taking anti-inflammatory supplements can be effective in improving lung cancer through reducing oxidative stress and inflammatory factors, though, this is not well known and needs to be further investigated. Therefore, we aimed to investigate the effect of 12 weeks of swimming training and royal jelly consumption on the expression of JNK and activator protein 1 (AP-1) genes in mice with Benzo[a]pyrene -induced lung cancer.
Materials and Methods: In this experimental study, 36 male Balb/c mice (age, 8 to 10 weeks; weight, 18 to 25 g) were randomly assigned to 6 groups: healthy control (n=6), sham (n=6), and experimental (n=24). Thereafter, cancer was induced by injection of 100 mg/kg body weight of benzopyrene, and the mice in the experimental group were divided into four subgroups of control with lung cancer (n=6), swimming training (n=6), royal jelly (n=6), and training + royal jelly (n=6). Swimming training was performed three times a week for 12 weeks. Royal jelly was also injected intraperitoneally at a dose of 100 mg/kg of body weight daily for 12 weeks. Forty-eight hours after the last training session, JNK and AP-1 gene expression were measured using Real-Time PCR. To analyze the data, a one-way analysis of variance (ANOVA) test with Tukey post-hoc tests and two-way ANOVA with Bonferroni post-hoc tests were used.
Results: Data analyses revealed that the expression of JNK and AP-1 genes in the lung cancer group was significantly increased compared to the healthy control group (p=0.001). Although the expression of JNK and AP-1genes were lower in the royal jelly group compared to the control cancer group (p=0.044, p=0.007, respectively), no significant differences were observed in the expression of these two genes between swimming training group and control cancer group (p=0.056, p=0.18, respectively). Finally, our result indicated that training + royal jelly group had the greatest decrease in expression of JNK and AP-1genes compared to the control cancer group (both p=0.001).
Conclusion: According to the study results, it seems that swimming training along with the consumption of royal jelly has more inhibitory and protective effects on the expression of JNK and AP-1 genes in lung tissue compared to either of these interventions in mice with lung cancer. To generalize the results to human, further studies on human subjects are warranted.

Keywords

Main Subjects

Sports Physiology (General)

References

1. Li C, Lei S, Ding L, Xu Y, Wu X, Wang H, et al. Global burden and trends of lung cancer incidence and mortality. Chinese Medical Journal. 2023;136(13):1583-1590
2. Leiter A, Veluswamy RR, Wisnivesky JP. The global burden of lung cancer: current status and future trends. Nature reviews Clinical oncology. 2-23;20(9):624-639
3. Doukas SG, Vageli DP, Lazopoulos G, Spandidos DA, Sasaki CT, Tsatsakis A. The Effect of NNK, A tobacco smoke carcinogen, on the miRNA and mismatch DNA repair expression profiles in lung and head and neck squamous cancer cells. Cells. 2023;9(4):1031.
4. Hu X, Geetha RV, Surapaneni KM, Veeraraghavan VP, Chinnathambi A, Alahmadi TA, et al. Lung cancer induced by Benzo(A)Pyrene: ChemoProtective effect of sinapic acid in swiss albino mice. Saudi Journal of Biological Sciences. 2021;28(12):7125–7133.
5. Shi H, Liu J, Gao H. Benzo (α) pyrene induces oxidative stress and inflammation in human vascular endothelial cells through AhR and NF-κB pathways. Microvascular Research. 2021; 137:104179.
6. El-Seedi HR, Eid N, Abd El-Wahed AA, Rateb ME, Afifi HS, Algethami AF, et al. Honey bee products: Preclinical and clinical studies of their anti-inflammatory and immunomodulatory properties. Frontiers in Nutrition. 2022; 8:761267.
7. Chen J, Ye C, Wan C, Li G, Peng L, Peng Y, et al. The roles of c-Jun N-terminal kinase (JNK) in infectious diseases. International Journal of Molecular Sciences. 2021;22(17):9640.
8. Qu F, Xu W, Deng Z, Xie Y, Tang J, Chen Z, et al. Fish c-Jun N-terminal kinase (JNK) pathway is involved in bacterial MDP-induced intestinal inflammation. Frontiers in Immunology. 2020; 11:459.
9. Song D, Lian Y, Zhang L. The potential of activator protein 1 (AP-1) in cancer targeted therapy. Frontiers in Immunology. 2023; 14:1224892.
10. Barzegari A, Mirdar S. Effect of 12-week submaximal swimming training in rats exposed to tobacco- derived nitrosamine ketone. Caspian Journal of International Medicine. 2018;9(2):158-163.
11. Luo Z, Wan R, Liu S, Feng X, Peng Z, Wang Q, et al. Mechanisms of exercise in the treatment of lung cancer - a mini-review. Frontiers in Immunology. 2023; 14:1244764.
12. Ge Z, Wu S, Qi Z, Ding S. Compared with high-intensity interval exercise, moderate intensity constant load exercise is more effective in curbing the growth and metastasis of lung cancer. Journal of Cancer. 2022;13(5):1468.
13. Gholamnezhad Z, Safarian B, Esparham A, Mirzaei M, Esmaeilzadeh M, Boskabady MH. The modulatory effects of exercise on lipopolysaccharide-induced lung inflammation and injury: A systemic review. Life Sciences. 2022; 293:120306.‏
14. Feng Y, Feng X, Wan R, Luo Z, Qu L, Wang Q. Impact of exercise on cancer: mechanistic perspectives and new insights. Frontiers in Immunology. 2024; 15:1474770.
15. Silva FCD, Iop RDR, Andrade A, Costa VP, Gutierres Filho PJB, Silva RD. Effects of physical exercise on the expression of microRNAs: A systematic review. Journal of Strength and Conditioning Research. 2020;34(1):270-280.
16. Gasparrini M. Afrin S, Forbes-Hernández TY, Cianciosi D, Reboredo-Rodriguez P, Amici A, et al. Protective effects of Manuka honey on LPS-treated RAW 264.7 macrophages. Part 2: Control of oxidative stress induced damage, increase of antioxidant enzyme activities and attenuation of inflammation. Food and chemical toxicology: an international journal published for the British Industrial Biological Research Association. 2018; 120:578-587.
17. Botezan S, Baci GM, Bagameri L, Pașca C, Dezmirean DS. Current status of the bioactive properties of royal jelly: A comprehensive review with a focus on its anticancer, anti-inflammatory, and antioxidant effects. Molecules (Basel, Switzerland). 2023;28(3):1510.
18. Sumartana IM, Setiaji Y. The physical and mental health benefits of swimming: Enhancing fitness, relaxation, endurance, and overall well-being. Journal of Interdisciplinary and Multidisciplinary Studies. 2025;1(2):88-100.
19. Abekova A, Islamov R. ChemoProtective effects of new iodine coordinated compound in benzo [a] pyrene-induced lung cancer in BALB/c mice. In BIO Web of Conferences. 2024; 100:3001.
20. Mirdar Harijani S, Musavi N. The effect of 12 weeks of submaximal swimming training on immunoreactivity of Ras and Raf-1 in lung epithelial cells of Wistar rats exposed to carcinogen NN. Research in Sport Medicine and Technology. 2020;18(19):113-126.
21. Mohammadi H, Mohammadian B, Najafzadeh Varzi H, Shahriari A. Evaluation of the effect of royal jelly on histopathological and biochemical changes in the lungs of rats following arsenic trioxide toxicity. Iranian Veterinary Journal. 2024;20(2):92-101.
22. Shirzad M, Kordyazdi R, Shahinfard N, Nikokar M. Does royal jelly affect tumor cells? Journal of HerbMed Pharmacology. 2013;2(2):45-48.
23. Ruiz EJ, Lan L, Diefenbacher ME, Riising EM, Da Costa C, Chakraborty A, et al. JunD, not c-Jun, is the AP-1 transcription factor required for Ras-induced lung cancer. JCI Insight. 2021;6(13): e124985.
24. Li C, Syed MU, Nimbalkar A, Shen Y, Vieira MD, Fraser C, et al. LKB1 regulates JNK-dependent stress signaling and apoptotic dependency of KRAS-mutant lung cancers. Nature Communications. 2025;16(1):4112
25. Cui L, Yang G, Ye J, Yao Y, Lu G, Chen J, et al. Dioscin elicits anti‐tumour immunity by inhibiting macrophage M2 polarization via JNK and STAT3 pathways in lung cancer. Journal of Cellular and Molecular Medicine. 2020;24(16):9217-9230.
26. Atsaves V, Leventaki V, Rassidakis GZ, Claret FX. AP-1 transcription factors as regulators of immune responses in cancer. Cancers. 2019;11(7):1037.
27. Bhosale PB, Kim HH, Abusaliya A, Vetrivel P, Ha SE, Park M. et al. Structural and functional properties of activator protein-1 in cancer and inflammation. Evidence Based Complementary and Alternative Medicine. 2022:9797929.
28. Brennan A, Leech JT, Kad NM, Mason JM. Selective antagonism of cJun for cancer therapy. Journal of Experimental and Clinical Cancer Research. 2020;39(1):184.
29. Ahmadi S, Taghian F. The Effect of eight weeks of intermittent training with quercetin encapsulated nanoparticles on JNK and ERK apoptosis regulators in the rat model of myocardial infarction. Iranian Journal of Endocrinology and Metabolism. 2021;23(2):133-142.
30. Kouhpayeh Z, Farsi S, Hoseini A, Fathi I. The effect of continuous and interval exercise with Crocin on the expression of AP-1 and NF-κB gene in the heart tissue of obese male Wistar rats. Journal of Applied Exercise Physiology, 2021;17(33):161-171.
31. Davari F, Alimanesh Z, Salehi O, Hosseini SA. Effect of training and crocin supplementation on mitochondrial biogenesis and redox-sensitive transcription factors in liver tissue of type 2 diabetic rats. Archives of Physiology and Biochemistry. 2022; 128(5):1215-1220.
32. Parker L, Trewin A, Levinger I, Shaw CS, Stepto NK. The effect of exercise-intensity on skeletal muscle stress kinase and insulin protein signaling. PloS One. 2017;12(2): e0171613.
33. Messina G, Tartaglia N, Ambrosi A, Porro C, Campanozzi A, Valenzano A, et al. The beneficial effects of physical activity in lung cancer prevention and/or treatment. Life (Basel, Switzerland). 2022;12(6):782.
34. Cui N, Li H, Dun Y, Ripley-Gonzalez JW, You B, Li D, et al. Exercise inhibits JNK pathway activation and lipotoxicity via macrophage migration inhibitory factor in nonalcoholic fatty liver disease. Frontiers in Endocrinology. 2022; 13:961231.‏
35. De Matos MA, de Oliveira Ottone V, Duarte TC, da Matta Sampaio PF, Costa KB, Fonseca CA, et al. Exercise reduces cellular stress related to skeletal muscle insulin resistance. Cell Stress and Chaperones. 2014;19(2):263-270.
36. Chen CW, Kuo YC, How CK, Juan CC. Long-term aerobic exercise training-induced anti-inflammatory response and mechanisms: Focusing on the toll-like receptor 4 signaling pathway. The Chinese Journal of Physiology. 2020;63(6):250-255.
37. Son ES, Park JW, Kim YJ, Jeong SH, Hong JH, Kim SH, Kyung SY. Effects of antioxidants on oxidative stress and inflammatory responses of human bronchial epithelial cells exposed to particulate matter and cigarette smoke extract. Toxicology in Vitro. 2020; 67:104883.
38. Kopustinskiene DM, Jakstas V, Savickas A, Bernatoniene J. Flavonoids as anticancer agents. Nutrients. 2020;12(2):457.
39. Albalawi AE, Althobaiti NA, Alrdahe SS, Alhasani RH, Alaryani FS, Mowyna MN. Antitumor activity of royal jelly and its cellular mechanisms against ehrlich solid tumor in mice. BioMed Research International. 2022;2022(1):7233997.
40. Lin XM, Liu SB, Luo YH, Xu WT, Zhang Y, Zhang T, et al. 10-HDA Induces ROS-Mediated Apoptosis in A549 Human Lung Cancer Cells by Regulating the MAPK, STAT3, NF-κB, and TGF-β1 Signaling Pathways. BioMed Research International. 2020;2020:3042636.
41. You MM, Chen YF, Pan YM, Liu YC, Tu J, Wang K, Hu FL. Royal jelly attenuates LPS‐induced inflammation in BV‐2 microglial cells through modulating NF‐κB and p38/JNK signaling pathways. Mediators of Inflammation. 2018;2018(1):7834381.
42. Ovchinnikov AN, Paoli A, Seleznev VV, Deryugina AV. Royal jelly plus coenzyme Q10 supplementation improves high-intensity interval exercise performance via changes in plasmatic and salivary biomarkers of oxidative stress and muscle damage in swimmers: a randomized, double-blind, placebo-controlled pilot trial. Journal of the International Society of Sports Nutrition. 2022;19(1):239-257.