Effect of Post-irradiation Inflammatory Response on Tight Junction Proteins in Rat Submandibular Glands

doi:10.13701/j.cnki.kqyxyj.2023.05.008

Abstract

Abstract: Objective: To observe the changes of tight junction (TJ) proteins induced by post-irradiation inflammation and its effect on paracellular pathway in submandibular glands (SMGs). Methods: Twenty-four rats were randomly divided into Control group and 1, 6, and 12 weeks post-irradiation group. The control group was not irradiated. In radiation group, a dose of 20 Gy irradiation was applied on the right SMGs. At specific time points, saliva secretion was measured within 5 min. The pathological changes were observed by HE staining. Immunohistochemical staining was used to investigate the positive expression of TNF-α. The levels and distribution of claudin-1, claudin-3, and AQP5 were detected by WB and immunofluorescence staining. The mRNA expression was detected by RT-qPCR. The levels of TNF-α, IL-1β, and IL-6 were detected by ELISA. Results: The salivary secretion was decreased after 1, 6, and 12 weeks in the irradiation group. The interstitial edema, degranulation, vacuolation, and inflammatory cell infiltration of SMGs were increased continuously. The levels of inflammatory cytokines were increased. The levels and mRNA expression of claudin-1, claudin-3, and AQP5 were down-regulated, and the relative fluorescence intensity was decreased. Conclusion: The inflammatory response after radiation may play a vital role affecting the abnormities of TJ proteins in rat SMGs and the destruction of paracellular pathway.

Key words: irradiation, inflammatory response, submandibular glands, paracellular pathway, tight junction proteins

ZHANG Pengxin, ABUDUNAIBI·Maimaitituerxun, WU Yanhui, AIHAITI·Guliziba, FEI Runxin, YAO Qingting, XU Hui. Effect of Post-irradiation Inflammatory Response on Tight Junction Proteins in Rat Submandibular Glands[J]. Journal of Oral Science Research, 2023, 39(5): 416-421.

References

[1] van der Laan HP, Van den Bosch L, Schuit E, et al. Impact of radiation-induced toxicities on quality of life of patients treated for head and neck cancer [J]. Radiother Oncol, 2021, 160: 47-53.
[2] Jasmer KJ, Gilman KE, Muñsoz Forti K, et al. Radiation-induced salivary gland dysfunction: Mechanisms, therapeutics and future directions [J]. J Clin Med, 2020, 9(12): 4095.
[3] Zhang XM, Huang Y, Zhang K, et al. Expression patterns of tight junction proteins in porcine major salivary glands: A comparison study with human and murine glands [J]. J Anat, 2018, 233(2): 167-176.
[4] Hosoi K, Yao C, Hasegawa T, et al. Dynamics of salivary gland AQP5 under normal and pathologic conditions [J]. Int J Mol Sci, 2020, 21(4): 1182.
[5] Bhat AA, Syed N, Therachiyil L, et al. Claudin-1, a double-edged sword in cancer [J]. Int J Mol Sci, 2020, 21(2): 569.
[6] Mei M, Xiang RL, Cong X, et al. Claudin-3 is required for modulation of paracellular permeability by TNF-α through ERK1/2/slug signaling axis in submandibular gland [J]. Cell Signal, 2015, 27(10): 1915-1927.
[7] 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.
[8] ukowski P, Maciejczyk M, Matczuk J, et al. Effect of N-acetylcysteine on antioxidant defense, oxidative modification, and salivary gland function in a rat model of insulin resistance [J]. Oxid Med Cell Longev, 2018, 2018: 6581970.
[9] Urek MM, Bralic M, Tomac J, et al. Early and late effects of X-irradiation on submandibular gland: a morphological study in mice [J]. Arch Med Res, 2005, 36(4): 339-343.
[10] Cong X, Zhang XM, Zhang Y, et al. Disruption of endothelial barrier function is linked with hyposecretion and lymphocytic infiltration in salivary glands of Sjögren's syndrome [J]. Biochim Biophys Acta Mol Basis Dis, 2018, 1864(10): 3154-3163.
[11] Min SN, Wu LL, Zhang YY, et al. Disruption of tight junction structure contributes to secretory dysfunction in IgG4-related sialadenitis [J]. J Mol Histol, 2020, 51(1): 33-46.
[12] Coppes R, Zeilstra L, Kampinga H, 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.
[13] Uchida H, Ingalls MH, Maruyama EO, et al. Short-term and bystander effects of radiation on murine submandibular glands [J]. Dis Model Mech, 2022, 15(11):dmm049570.
[14] Araujo M, Spadella M, Chies A, et al. Effect of low radiation dose on the expression and location of aquaporins in rat submandibular gland [J]. Tissue Cell, 2018, 53: 104-110.
[15] López-Villalobos EF, Muñoz-Valle JF, Palafox-Sánchez CA, et al. Cytokine profiles and clinical characteristics in primary Sjögren's syndrome patient groups [J]. J Clin Lab Anal, 2021, 35(2): e23629.
[16] Limaye A, Hall B, Zhang L, et al. Targeted TNF-α overexpression drives salivary gland inflammation [J]. J Dent Res, 2019, 98(6): 713-719.
[17] Yoshimoto S, Yoshizumi J, Anzai H, et al. Inhibition of Alk signaling promotes the induction of human salivary-gland-derived organoids [J]. Dis Model Mech, 2020, 13(9): dmm045054.
[18] Mei M, Qu LH, Cong X, et al. CTRP3 promotes TNF-α-induced apoptosis and barrier dysfunction in salivary epithelial cells [J]. Cell Signal, 2021, 85: 110042.
[19] Kawedia JD, Nieman ML, Boivin GP, et al. Interaction between transcellular and paracellular water transport pathways through Aquaporin 5 and the tight junction complex [J]. Proc Natl Acad Sci U S A, 2007, 104(9): 3621-3626.
[20] Ewert P, Aguilera S, Alliende C, et al. Disruption of tight junction structure in salivary glands from Sjögren's syndrome patients is linked to proinflammatory cytokine exposure [J]. Arthritis Rheum, 2010, 62(5): 1280-1289.
[21] Zhang XM, Huang Y, Cong X, et al. Parasympathectomy increases resting secretion of the submandibular gland in minipigs in the long term [J]. J Cell Physiol, 2019, 234(6): 9515-9524.
[22] Zhou J, Yu Q. Anti-IL-7 receptor-α treatment ameliorates newly established Sjögren's-like exocrinopathy in non-obese diabetic mice [J]. Biochim Biophys Acta Mol Basis Dis, 2018, 1864(7): 2438-2447.