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1.
Oral Dis ; 28(6): 1528-1538, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33818901

RESUMO

OBJECTIVES: To investigate the effects of radiation on paracellular pathway of rat submandibular glands (SMGs) and the mechanism of increasing secretion following treatment with pilocarpine. MATERIALS AND METHODS: In situ irradiation models of SMGs in Wistar rats were conducted, and the glands were exposed to X-radiation at a single dose of 20 Gy. Pilocarpine was intraperitoneally injected 60 min prior to radiation and continuous 6 days postirradiation for a total of 7 days. Salivary secretion, histological changes, pro-inflammatory cytokines, alterations in tight junctions (TJs), and functional membrane proteins aquaporin-5 (AQP5) and claudin-4 mediated by the muscarinic acetylcholine M3 subtype receptor were determined at 1 and 12 weeks after irradiation. RESULTS: Salivary secretion of the irradiated glands was reduced at 1 and 12 weeks. As well, acinar cell numbers, TJ width, and the levels of M3 receptor and AQP5 were decreased. In contrast, tumor necrosis factor-α, interleukin 6, interleukin 1α, and the expression of the TJ protein claudin-4 were significantly increased in irradiated SMGs. Notably, all the alterations were attenuated by pilocarpine treatment. CONCLUSIONS: Pilocarpine could improve the secretory function of irradiated rat SMGs via reducing inflammation, ameliorating the structural injury of TJs, and attenuating the up-regulation of claudin-4 expression.


Assuntos
Pilocarpina , Glândula Submandibular , Animais , Claudina-4/metabolismo , Claudinas/metabolismo , Claudinas/farmacologia , Pilocarpina/farmacologia , Ratos , Ratos Wistar , Junções Íntimas/metabolismo
2.
Shanghai Kou Qiang Yi Xue ; 31(4): 359-366, 2022 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-36710547

RESUMO

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 salivarysecretion 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.


Assuntos
Claudinas , Glândula Parótida , Ratos , Masculino , Animais , Claudinas/metabolismo , Claudina-4/metabolismo , Junções Íntimas/metabolismo , Ratos Wistar , Radiação Ionizante
3.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 39(3): 267-273, 2021 Jun 01.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-34041874

RESUMO

OBJECTIVES: This study aims to investigate the effects of ionizing radiation on the secretion of the paracellular pathway in rat submandibular glands (SMGs) and reveal the changes in the tight junction (TJ) protein claudin-4. METHODS: A total of 24 Wistar rats were randomly divided into control and irradiation groups. The irradiation groups were further divided into 1, 4, and 12 weeks groups after irradiation. One-time 20 Gy irradiation was given to the SMG area on the experimental side of the irradiation group. At 1, 4, and 12 weeks after irradiation, the secretion of SMGs was measured using the Schirmer's test. The pathological changes in the gland tissues were observed under light microscopy after hematoxylin⁃eosin (HE) staining. The changes in the TJ ultrastructure were observed under transmission electron microscopy. The immunofluorescence staining and Western blot were used to detect the expression levels of muscarinic acetylcholine M3 receptor, aquaporin 5 (AQP5), and claudin-4 protein. RESULTS: At 1, 4, and 12 weeks after irradiation, the secretion of SMGs in the irradiation group was significantly decreased and lower than that in the control group (P<0.01). At 1 week, the interstitial edema was observed in SMG tissues. Nuclear pyknosis, decreased number of acinar cells, and small focal necrosis with inflammatory infiltration were also observed over time. However, these changes were most evident at 12 weeks after irradiation. In the irradiation group, the TJ ultrastructure of glands at different times appeared to be fuzzy, collapsed, and had decreased electron density. Moreover, the width of TJs was remarkably decreased (P<0.01). The expression levels of M3 and AQP5 were decreased in a time-dependent manner, and the fluorescence intensity was significantly reduced after irradiation. However, the expression levels and fluorescence intensity of claudin-4 were enhanced in different degrees. CONCLUSIONS: The changes in the TJ structure, the upregulation of the claudin-4 expression, and the damage in the paracellular pathway were involved in the hyposecretion of SMGs after irradiation.


Assuntos
Glândula Submandibular , Junções Íntimas , Animais , Microscopia Eletrônica de Transmissão , Radiação Ionizante , Ratos , Ratos Wistar
4.
Eur J Oral Sci ; 129(3): e12785, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33786924

RESUMO

To investigate the effects of radiation on rat submandibular glands and the possible protective effects of ischemic preconditioning, the submandibular glands of Wistar rats were subjected to in situ radiation after ischemic preconditioning. The glands were exposed to X-radiation at a single dose of 20 Gy. Ischemic preconditioning was achieved by three min of ischemia and three min of reperfusion, repeated three times before irradiation. Salivary secretion, histological changes, alterations in tight junctions, and the levels of oxidative stress, pro-inflammatory cytokines, and water secretion proteins mediated by the muscarinic acetylcholine M3 subtype receptor were determined at 1 and 12 weeks post-irradiation. In glands subjected to irradiation only, the secretion, superoxide dismutase activity, tight junction width, acinar cell number, and M3 receptor and aquaporin-5 levels were lower at 1 and 12 weeks than seen in the ischemically preconditioned irradiated glands. In contrast, tumor necrosis factor-α, malondialdehyde, myeloperoxidase activity, and the expression of the tight junction protein claudin-4 were significantly higher in the irradiated only glands. Our study revealed that radiation caused a series of injury-stress responses, especially damage to the water secretion pathway mediated by the M3 receptor that ultimately led to hyposecretion, which might play an important role in the dysfunction of the irradiated only glands. Ischemic preconditioning reduced the radiation-induced injury to submandibular glands and ameliorated salivary hyposecretion.


Assuntos
Precondicionamento Isquêmico , Glândula Submandibular , Animais , Ratos , Ratos Wistar , Receptor Muscarínico M3 , Salivação
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