电离辐射对大鼠腮腺旁细胞分泌途径损伤及紧密连接蛋白claudin-4的影响

doi:10.19439/j.sjos.2022.04.005

上海口腔医学 ›› 2022, Vol. 31 ›› Issue (4): 359-366.doi: 10.19439/j.sjos.2022.04.005

电离辐射对大鼠腮腺旁细胞分泌途径损伤及紧密连接蛋白claudin-4的影响

姚清婷^1,2, 吴言辉^1,2, 刘少华^3,4, 许辉¹, 李军¹, 石亮^3,4

1.新疆维吾尔自治区人民医院口腔颌面外科,新疆乌鲁木齐 830001;
2.新疆医科大学,新疆乌鲁木齐 830054;
3.山东大学齐鲁医院口腔颌面外科,山东济南 250012;
4.山东大学口腔医学研究所,山东济南 250012

收稿日期:2021-01-28 修回日期:2021-04-17 出版日期:2022-08-25 发布日期:2022-08-30
通讯作者: 石亮,E-mail：shiliang1976@126.com
作者简介:姚清婷 (1993-),女,硕士研究生,E-mail：yaoqingting@126.com
基金资助:
山东省重点研发计划项目（GG201709220011）; 山东省自然科学基金（ZR2020MH187）

Damage on paracellular pathway and effect on tight junction protein claudin-4 in rat parotid glands caused by ionizing radiation

YAO Qing-ting^1,2, WU Yan-hui^1,2, LIU Shao-hua^3,4, XU Hui¹, LI Jun¹, SHI Liang^3,4

1. Department of Oral and Maxillofacial Surgery, People's Hospital of Xinjiang Uygur Autonomous Region. Urumqi 830001, Xinjiang Uygur Autonomous Region;
2. Xinjiang Medical University. Urumqi 830054, Xinjiang Uygur Autonomous Region;
3. Department of Oral and Maxillofacial Surgery, Qilu Hospital of Shandong University. Jinan 250012, Shandong Province;
4. Institute of Stomatology, Shandong University. Jinan 250012, Shandong Province, China

Received:2021-01-28 Revised:2021-04-17 Online:2022-08-25 Published:2022-08-30

摘要/Abstract

摘要： 目的: 探讨电离辐射对大鼠腮腺旁细胞分泌功能损伤及紧密连接 (tight junction,TJ) 蛋白claudin-4的影响与潜在作用机制。方法: 24只8周龄雄性Wistar大鼠,随机分为对照组（6只）和照射组（照射后1周组、4周组、12周组,每组6只）。照射组一次性20 Gy射线局部照射实验侧腮腺区。采用Schirmer实验,检测各组腺体唾液静息分泌量;H-E染色,光镜下观察腺体组织病理变化;透射电镜观察TJ超微结构改变;免疫荧光染色和蛋白质印迹检测毒蕈碱型乙酰胆碱受体（M受体）亚型M3、水通道蛋白5（AQP5）及claudin-4的蛋白表达。采用SPSS 23.0软件包对数据进行统计学分析。结果: 照射后1、4、12周,腺体静息分泌量较对照组显著减少（P<0.05）;12周较1周、4周时减少更显著（P<0.05）。组织学观察可见,照射后腺体早期间质血管扩张、充血,后期腺泡细胞计数显著减少（P<0.05）;TJ结构模糊、电子密度降低、宽度减少（P<0.05）。免疫荧光染色和蛋白质印迹检测结果显示,M3受体及AQP5蛋白在照射后1、4、12周表达显著下调,而claudin-4蛋白表达显著升高。结论: 电离辐射后腮腺旁细胞分泌功能降低,TJ结构改变,claudin-4表达上调,可能参与腺体分泌功能损伤。

关键词: 电离辐射, 紧密连接, Claudin-4, 旁细胞途径, 腮腺

Abstract: PURPOSE: To investigate the effect and the potential mechanism of ionizing radiation on secretory function and tight junction (TJ) protein claudin-4 in paracellular pathway of rat parotid glands. METHODS: Twenty four 8-week-old male Wistar rats were randomly divided into one control group(n=6) and three irradiation groups (i.e., 1-week group post-irradiation, 4-week group post-irradiation, and 12-week group post-irradiation, 6 rats in each group). The experimental glands of irradiation groups were exposed to X-radiation in one-time single doses of 20 Gy. The residual salivary secretion of parotid glands was measured by Schirmer's test. The pathological changes of gland tissues were observed under light microscope after hematoxylin-eosin(H-E) staining. The changes of TJs ultrastructure were observed under transmission electron microscopy (TEM). Immunofluorescence staining and Western blot were used to detect the expression levels of muscarinic acetylcholine M3 subtype receptor, aquaporin 5 (AQP5), and claudin-4 proteins. The experimental results were analyzed with SPSS 23.0 software package for one-way analysis of variance. RESULTS: The residual salivary secretion of irradiation group glands at 1, 4, and 12 weeks after irradiation was reduced compared with that of the control group(P<0.05), and the residual salivary secretion of irradiation group at 12 weeks was significantly lower than that at 4 weeks(P<0.05). Histologically, the dilation and congestion of interstitial vessels were observed at early stage after irradiation, and significant reduced number of acinar cells was found at late stage(P<0.05). In the irradiation groups, the ultrastructures of TJ were fuzzy, the electron density was decreased, and the TJ width at 1, 4, and 12 weeks was reduced compared with that in the control group. Immunofluorescence staining and Western blot indicated that the protein expression levels of M3 and AQP5 were down-regulated; however, the protein expression levels of claudin-4 were significantly increased at 1, 4, and 12 weeks after irradiation. CONCLUSIONS: After ionizing radiation, decreased secretory function of paracellular pathway, alterations in TJ structures, and up-regulation of claudin-4 expression may be involved in the mechanism of hyposecretion in rat parotid glands after irradiation.

Key words: Ionizing radiation, Tight junction, Claudin-4, Paracellular pathway, Parotid gland

中图分类号:

R739.8

姚清婷, 吴言辉, 刘少华, 许辉, 李军, 石亮. 电离辐射对大鼠腮腺旁细胞分泌途径损伤及紧密连接蛋白claudin-4的影响[J]. 上海口腔医学, 2022, 31(4): 359-366.

YAO Qing-ting, WU Yan-hui, LIU Shao-hua, XU Hui, LI Jun, SHI Liang. Damage on paracellular pathway and effect on tight junction protein claudin-4 in rat parotid glands caused by ionizing radiation[J]. Shanghai Journal of Stomatology, 2022, 31(4): 359-366.

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电离辐射对大鼠腮腺旁细胞分泌途径损伤及紧密连接蛋白claudin-4的影响

Damage on paracellular pathway and effect on tight junction protein claudin-4 in rat parotid glands caused by ionizing radiation

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