Effect of Oxidative Stress after Radiation on Apoptosis of Cells in Rats Submandibular Gland

doi:10.13701/j.cnki.kqyxyj.2024.07.007

Abstract

Abstract: Objective: To observe the effect of oxidative stress induced by submandibular gland (SMG) radiation on apoptosis of acinar cells in rats. Methods: Twenty-four rats were randomly divided into control group, 24 h, 72 h, and 1 w group. The control group was not irradiated, the right SMG of the radiation group rats was received one-time 20 Gy radiation, and the saliva secretion of each group was detected. The tissue morphology of SMG was observed by HE staining. The levels of malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD) were detected by ELISA. The positive expression of 8-OHdG was observed by IHC staining. The apoptosis of SMG cells was detected by TUNEL staining. The levels of apoptosis-related proteins BCL2-associated X protein (BAX), B-cell lymphoma-2 (Bcl-2), and cysteinyl aspartate specific proteinase-3 (Caspase-3) were detected by western blot, and the mRNA expression was detected by RT-qPCR. Results: Salivary secretion was decreased after radiation. The pathological changes such as intracellular vacuolation, tissue edema, and gap widening were gradually increased. The cells showed obvious oxidative stress and apoptosis. The expression levels of apoptosis-related proteins BAX, Bcl-2, and Caspase-3 were abnormal. Conclusion: Post-radiation oxidative stress may be an important factor affecting apoptosis and dysfunction of SMG cells.

Key words: radiation damage, submandibular gland, oxidative stress, apoptosis

Maimaitituerxun·ABUDUNAIBI, Rezeye·MAIMAITIZUNONG, ZHANG Pengxin, QI Jia, WU Shihan, XU Hui. Effect of Oxidative Stress after Radiation on Apoptosis of Cells in Rats Submandibular Gland[J]. Journal of Oral Science Research, 2024, 40(7): 605-610.

References

[1] Johnson DE, Burtness B, Leemans CR, et al. Head and neck squamous cell carcinoma [J]. Nat Rev Dis Primers, 2020, 6(1): 92.
[2] Mody MD, Rocco JW, Yom SS, et al. Head and neck cancer [J]. Lancet, 2021, 398(10318): 2289-2299.
[3] Luitje ME, Israel AK, Cummings MA, et al. Long-term maintenance of acinar cells in human submandibular glands after radiation therapy [J]. Int J Radiat Oncol Biol Phys, 2021, 109(4): 1028-1039.
[4] 李审绥,吴沉洲,乔翔鹤,等. 辐射损伤唾液腺机制及治疗的研究进展[J].华西口腔医学杂志,2021,39(1):99-104.
[5] Coppes RP, Vissink A, Konings AWT. Comparison of radiosensitivity of rat parotid and submandibular glands after different radiation schedules [J]. Radiother Oncol, 2002, 63(3): 321-328.
[6] Li X, Fang F, Gao Y, et al. ROS induced by killerred targeting mitochondria (mtKR) enhances apoptosis caused by radiation via Cyt c/Caspase-3 pathway [J]. Oxid Med Cell Longev, 2019, 2019:4528616.
[7] Wei J, Wang B, Wang H, et al. Radiation-induced normal tissue damage: Oxidative stress and epigenetic mechanisms [J]. Oxid Med Cell Longev, 2019, 2019:3010342.
[8] Kim JH, Kim KM, Jung MH, et al. Protective effects of alpha lipoic acid on radiation-induced salivary gland injury in rats [J]. Oncotarget, 2016, 7(20): 29143-29153.
[9] Wu YH, Yao QT, Liu SH, et al. Effect of ischemic preconditioning on radiation damage to the submandibular gland in rats [J]. Eur J Oral Sci, 2021, 129(3): e12785.
[10] Bock FJ, Tait SWG. Mitochondria as multifaceted regulators of cell death [J]. Nat Rev Mol Cell Biol, 2020, 21(2): 85-100.
[11] Coppes RP, Zeilstra LJ W, Kampinga HH, et al. Early to late sparing of radiation damage to the parotid gland by adrenergic and muscarinic receptor agonists [J]. Br J Cancer, 2001, 85(7): 1055-1063.
[12] 张鹏鑫,麦麦提吐尔逊·阿布都乃比,吴言辉,等. 放射后炎症反应对大鼠颌下腺紧密连接蛋白的影响[J].口腔医学研究,2023,39(5):416-421.
[13] Zhang C, Xiang B. The underlying mechanisms and strategies of DNA damage and repair in radiation sialadenitis [J]. Oral Dis, 2023, 29(3): 990-995.
[14] Jasmer KJ, Gilman KE, Muñoz Forti K, et al. Radiation-induced salivary gland dysfunction: Mechanisms, therapeutics and future directions [J]. J Clin Med, 2020, 9(12):4095.
[15] Prasad S, Gupta SC, Tyagi AK. Reactive oxygen species (ROS) and cancer: Role of antioxidative nutraceuticals [J]. Cancer Lett, 2017, 387:95-105.
[16] Liu X, Subedi KP, Zheng C, et al. Mitochondria-targeted antioxidant protects against irradiation-induced salivary gland hypofunction [J]. Sci Rep, 2021, 11(1): 7690.
[17] Yao QT, Wu YH, Liu SH, et al. Pilocarpine improves submandibular gland dysfunction in irradiated rats by downregulating the tight junction protein claudin-4 [J]. Oral Dis, 2022, 28(6): 1528-1538.
[18] Jelic MD, Mandic AD, Maricic SM, et al. Oxidative stress and its role in cancer [J]. J Cancer Res Ther, 2021, 17(1): 22-28.
[19] Graille M, Wild P, Sauvain JJ, et al. Urinary 8-OHdG as a biomarker for oxidative stress: A systematic literature review and meta-analysis [J]. Int J Mol Sci, 2020, 21(11):3743.
[20] Ren J, Huang R, Li Y, et al. Radioprotective effects and mechanism of HL-003 on radiation-induced salivary gland damage in mice [J]. Sci Rep, 2022, 12(1): 8419.
[21] Perillo B, Di Donato M, Pezone A, et al. ROS in cancer therapy: The bright side of the moon [J]. Exp Mol Med, 2020, 52(2):192-203.
[22] Matsumoto N, Omagari D, Ushikoshi-Nakayama R, et al. Hyperglycemia induces generation of reactive oxygen species and accelerates apoptotic cell death in salivary gland cells [J]. Pathobiology, 2021, 88(3):234-241.
[23] Wang XY, Liu KJ, Zhang FY, et al. Nicotinamide mitigates radiation injury in submandibular gland by protecting mitochondrial structure and functions [J]. J Oral Pathol Med, 2022, 51(9): 801-809.
[24] Meng DF, Guo LL, Peng LX, et al. Antioxidants suppress radiation-induced apoptosis via inhibiting MAPK pathway in nasopharyngeal carcinoma cells [J]. Biochem Biophys Res Commun, 2020, 527(3): 770-777.
[25] Morgan MJ, Liu Z. Crosstalk of reactive oxygen species and NF-κB signaling [J]. Cell Res, 2011, 21(1): 103-115.
[26] 曹牛奔,刘笑梦,邓愉,等. 活性氧/c-Jun氨基末端激酶/核因子-κB信号分子通过调控凋亡参与牙周炎诱导肝损伤[J].华西口腔医学杂志,2022,40(5):532-540.