• Journal of Oral Medicine and Pain
• /
• v.38 no.4
• /
• pp.291-298
• /
• 2013

On this study, we treated rats with restraint stress, and observed the changes with an optical microscope. Within the salivary gland tissue, we measured cell apoptosis cycle evaluation which show positive reaction on TUNEL assay, and compared within the groups. For this study, 18 rats were divided into 3 groups; 1) 2 rats of group I were selected as a normal control. 2) 2 rats of group II, as a experimental control were placed in the restraint cone for 2 hours 3) 14 rats of group III were placed in the restraint cone for 2 hours once a day. The rats were sacrificed immediately (group II, as a experimental control), 1, 2, 3, 4, 5, 6 and 7days after application of the stress and the both parotid glands were excised. The conclusions follow. 1. 5 days after giving an confining stress to the parotid gland of Rats, we can observe the hypotropy and pus and inflammation of Rat parotid gland acinar cells, and after 7 days, we can see a cell apoptosis. 2. Through the In situ DNA end labeling assay and TUNEL dye, on serous glands, benign tumor cell increased with statistically significant result after 5 days from confining stress. And the index shows maximum value on 7th days, which is same result with histological opinion. Therefore, our study shows that a cell apoptosis can be induced by restraint stress on salivary gland tissue, and we think more study should be accomplished about the cell signaling pathway in the future.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.30 no.2
• /
• pp.85-99
• /
• 2004

Purpose: This study observed the changes in the telomerase activity, it's developmental regulation, PCNA expression, and their correlation in rat's upper digestive organs during growth and aging. Materials and Methods: Upper digestive organs(buccal mucosa, gingiva, palate, submandibular and parotid glands, and tongue) were aseptically removed from Sprague-Dawley rats of fetal(gestational 20 days), growing(1, 2, 3, 5, and 7 weeks after birth) and adult(12 week old). Samples for telomerase activity were frozen on liquid nitrogen immediately after sacrifice, and stored until the use at $-75^{\circ}C$ in order to measure it. Telomerase activity was measured by a PCR-based telomeric repeat amplication protoco(TRAP) assay and quantitated with Photometric Telo TAGGG Telomerase PCR ELISA plus(Roche Diagnostics GmbH. Mannheim. Germany). PCNA expression were measured immunohistochemistry with anti PCNA Ab-1, Clone PC10(NeoMark. California. USA). Results: 1. Telomerase activities in buccal mucosa, palate and gingiva were the highest in fetus and decreased gradually or rapidly after birth and then diminished, but In salivary gland and tongue were the highest in fetus and also high at 1 week and then decreased rapidly. 2. PCNA expression in buccal mucosa, gingiva, Tongue and salivary gland was the highest in fetus and decreased gradually and then diminished. but only in palate decreased rapidly after birth and then diminished. Conclusion: The highest telomerase activity of embryonic stage decreased rapidly after birth in rat's upper digestive organs. There may be a developmental regulation of telomerase activity, but not a tissue-specific. This telomerase activity seems correlated closely with PCNA expression in rat's upper digestive system.

• The Korean Journal of Physiology and Pharmacology
• /
• v.7 no.2
• /
• pp.97-101
• /
• 2003

The salivary glands produce 1.5L of fluid per day. As in other exocrine organs, the general mechanism in the salivary glands is that water movement occurs secondary to osmotic driving forces created by active salt transport. Therefore, high water permeability in the salivary glands is expected to have a variety of aquaporin (AQP), a water channel. Although some AQPs have been known to be present in the salivary glands, roles of parasympathetic nerve in AQP expression have not yet been examined. This study was designed to examine the changes of AQPs and extracellular signal-regulated kinase (ERK) in the submandibular glands after parasympathetic denervation. Right chorda-lingual nerve was cut, and each right (experiment) and left (control) submandibular gland was excised at 1, 3, 7, 14, 30 days after denervation. The denervated right submandibular glands were resulted in weight loss and morphologic changes, including cell loss and atrophy, as the time elapsed after parasympathetic denervation increased, whereas there were no histologic alteration in control side. AQP5 which is known to reside in apical membrane and secretory caraliculi of the submandibular acini were gradually underexpressed according, as the time after denervation increased. Expression of AQP4 in submandibular ductal epithelium was down-regulated after denervation. Besides, AQP3 and 8, which is known to be present in basolateral membrane of the glandular acini, were gradually underexpressed after denervation, similar to the pattern of other types. Expression of ERK, a mitogen-activated protein kinase, was downregulated after parasympathetic denervation in the submandibular gland. These results suggest that parasympathetic nervous system regulates the expression of AQPs in salivary glands, and is in part mediated by ERK pathway.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.31 no.1
• /
• pp.24-30
• /
• 2005

The flow of saliva is controlled entirely by nervous stimuli. The present study was aimed to explore the role of sympathetic and parasympathetic nerves in the regulation of sodium transporters and water channels in the salivary gland. Rats were denervated of their sympathetic and parasympathetic nerves to the submandibular gland, and the expression of sodium transporters and water channels was determined. The expression of either ${\alpha}-1$ or ${\beta}-1$ subunit of Na, K-ATPase was not significantly affected by the sympathetic denervation. On the contrary, the expression of both subunits was decreased by the parasympathetic denervation. The expression of ${\alpha}-,\;{\beta}-$, and ${\gamma}$-subunits of ENaC was not significantly affected by the sympathetic denervation, but was increased by the parasympathetic denervation. On the contrary, the expression of NHE3 was markedly decreased by both the sympathetic and the parasympathetic denervation. The sympathetic denervation significantly increased the expression of AQP1, while the parasympathetic denervation was without effect. The sympathetic and parasympathetic denervation significantly increased the expression of AQP4. The sympathetic denervation did not affect the expression of AQP5, but the parasympathetic denervation significantly decreased it. These results suggest that sympathetic and parasympathetic nerves have tonic effects on the regulation of sodium transporters and AQP water channels in the salivary gland. The sympathetic and parasympathetic denervation may then result in alterations of secretory rate and electrolyte composition of the saliva.

• International Journal of Oral Biology
• /
• v.37 no.4
• /
• pp.153-159
• /
• 2012

The effects of the an immunosuppressive drug cyclosporine A (CsA), on the salivary gland are largely unknown, even though clinical trials for the stimulation of salivation using CsA have been attempted. Cyclophilin A (CypA) is known to be a binding protein for CsA. CypA has cell proliferation and tissue matrix change activities. In our present study, the presence of CypA in the gland and effects of CsA on CypA expression were investigated by immunohistochemistry, immunoblotting and RT-PCR analyses. CypA was immunohistochemically detected in various kinds of ducts in the submandibular glands of Sprague Dawley rats. The CypA mRNA level was highest at postnatal day 1 and gradually decreased in a time-dependent manner up to adulthood. The expression of CypA increased after a 10 day subcutaneous administration of CsA in postnatal day 1 rats. Surgical sections of the chorda-lingual nerve with impaired salivation showed no changes in CypA expression. A cell proliferation assay using PCNA anti-serum showed increased cell division following CsA treatment. These results suggest that CsA and CypA may act on ductal cells to regulate saliva composition rather than salivation levels.

• The Korean Journal of Physiology and Pharmacology
• /
• v.6 no.6
• /
• pp.305-309
• /
• 2002

There are some evidences that $K^+$ efflux evoked by muscarinic stimulation is not mainly mediated by large conductance $K^+$ (BK) channels in salivary gland. In this experiment, we therefore characterised non BK channels in rat submandibular gland acinar cells and examined the possibility of agonist effect on this channel using a patch clamp technique. Two types of $K^+$ channels were observed in these cells. BK channels were observed in 3 cells from total 6 cells and its average conductance was $152{\pm}7$ pS (n=3). The conductance of the another types of $K^+$ channel was estimated as $71{\pm}7$ pS (n=6). On the basis of the conductance of this channel, we defined this channel as intermediate conductance $K^+$ (IK) channels, which were observed from all 6 cells we studied. When we increased $Ca^{2+}$ concentration of the bath solution in inside-out mode, the IK channel activity was greatly increased, suggesting this channel is $Ca^{2+}$ sensitive. We next examined the effect of carbachol (CCh) and isoproterenol on the activity of the IK channels. $10^{-5}$ M isoproterenol significantly increased the open probability (Po) from $0.08{\pm}0.02$ to $0.21{\pm}0.03$ (n=4, P<0.05). Application of $10^{-5}$ M CCh also increased Po from $0.048{\pm}0.03$ to $0.55{\pm}0.33$ (n=5, P<0.05) at the maximum channel activity. The degree of BK channel activation induced by the same concentration of CCh was lower than that of IK channels; Po value was $0.011{\pm}0.003$ and $0.027{\pm}0.005$ in control and during CCh stimulation (n=3), respectively. The result suggests that IK channels exist in salivary acinar cells and its channel activity is regulated by muscaricinic and ${\beta}-adrenergic$ agonist. We conclude that IK channels also play a putative role in secretion as well as the BK channels in rat submandibular gland acinar cells.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
• /
• v.16 no.1
• /
• pp.49-58
• /
• 1986

This study was designed to investigate the irradiation effects on the rat parotid gland, applied to the head and neck region. For this experiment, twenty-four rats, feeded under the even condition, were used as experimental animals. Twenty rats were used for experimental group and the rest were assigned to the control group. The experimental group was singly irradiated with 10Gray through Cobalt-60 radiotherapy device, Picker model 4M 60 (Field size; l2×5㎝, SSD; 50㎝, Depth; 1㎝). The experimental animals of both group were sacrificed each four animals in 2 days, 1week, 2weeks, 3weeks and 4weeks after irradiation. The specimens were examined through the light microscope using the H-E stain and H stain by routin procedure. The other specimens were observed under the fluorescence microscope using the B-O dichroic mirror and Y455 barrier filter after PA-ACH stain. 1. The results of this study were obtained as follows, The parotid acini were severely degenerated and the intraacinar spaces were widened. Within the acini, retained secretory granules and increased fibrosis were observed. Also the shape and the size of the acini showed very irregular atrophic degenerations. 2. The nuclei showed severe pyknosis, displacement and irregular aggregated appearance. 3. The tissue changes of the parotid acini were initiated after 2 days of irradiation and most severely appeared at the second week of irradiation, but almost returned to normal. 4. The salivary ducts of the parotid gland were severely atrophied, discontinued but initiated to regenerated after 3 weeks of irradiation.

• Radiation Oncology Journal
• /
• v.17 no.3
• /
• pp.230-237
• /
• 1999

Purpose : To elucidate the effects of pentoxifylline and diltiazem on the late response of the salivary glands of the rat after irradiation. Materials and Methods : Sixteen Sprague-Dawley rats were divided into 4 groups : (a) irradiation alone (b) irradiation with pentixifylline (PTX) (c) irradiation with diltiazem (DTZ) (d) irradiation with both PTX and DTZ. Irradiation was given in a single fraction of 16 Gy using 4 MV photon energy through an anterior port encompassing the left side of the salivary gland leaving the right side of salivary gland as a control. PTX, 20 mg/kg and/or DTZ, 50 mg/kg were infused intraperitoneally before irradiation, Two rats from each group were sacrificed on the 10th week and the rest was sacrificed on the 16th week after irradiation. Histopathologic examinations were undertaken for each section and the proportion of vacuolated cells out of the total number of cells under light microscopic fields was calculated. The statistical significance in the difference of the proportion of the vacuolated cells among the experimental groups was evaluated by a $x^2$-test. Results : Irradiated salivary glands of the 10th week group revealed markedly increased number of vacuolated cells compared to those of unirradiated control. The proportion of vacuolated cells was significantly reduced in both the PTX group (p value=0.001) and the combined PTX and DTX group compared to those of irradiation alone group. The DTZ alone group did not reveal the significant reduction of vacuolated cells compared to those of irradiation alone group (p value, >0.05). The 16th week groups revealed similar findings to those of the 10th week group, but the degree of chronic inflammatory cell infiltrates and interstitial fibrosis was increased and the number of acinar cells was reduced compared to those of the 10th week group. Conclusions : PTX significantly reduced the late radiation response of salivary glands, but DTZ did not reduce the same degree as PTX did. Taking the positive results of this study into consideration, it seems reasonable to apply PTX into the clinical trial for the head and neck irradiation to reduce the late radiation sequelae of salivary glands in the near future. At the same time the further experiment to clarify the subcellar mechni는 involved in PTX should be preceded.

• Imaging Science in Dentistry
• /
• v.33 no.3
• /
• pp.161-169
• /
• 2003

Purpose: To observe the histologic changes and clusterin expression in the acinar cells of the submandibular gland in streptozotocin-induced diabetic rat following irradiation. Materials and Methods: Mature Sprague-Dawley rats were divided into three groups: control, diabetic, and diabetic-irradiated groups. Diabetes mellitus was induced in the Sprague-Dawley rats by injecting streptozotocin, while the control rats were injected with citrate buffer only. After 5 days, rats in diabetic-irradiated group were irradiated with single absorbed dose of 10 Gy to the head and neck region. The rats were killed at 1, 3, 7, 14,21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histologic and immunohistochemical methods. Results : Morphologic change of acinar cells was remarkable in the diabetic group, but was not observed in the diabetic-irradiated group. Necrotic tissues were observed in the diabetic-irradiated group. Coloring of toluidine blue stain was most increased at 14 days in the diabetic group, however there were no significant change throughout the period of the experiment in the diabetic-irradiated group. Expression of clusterin was most significant at 14 days in the diabetic group, but gradually decreased with time after 7 days in the diabetic-irradiated group. Degeneration of clusterin was observed in the diabetic-irradiated group. Conclusion : This experiment suggests that the acinar cells of submandibular gland in rats are physiologically apoptosed by the induction of diabetes, but that the apoptosis is inhibited and the acinar cells necrotized after irradiation.

• Imaging Science in Dentistry
• /
• v.36 no.1
• /
• pp.33-40
• /
• 2006

Purpose : To investigate clusterin expression in the acini and ductal cells of rat submandibular glands after Co-60 gamma irradiation. Materials and Methods : The male Sprague-Dawley rats weighing approximately 250 gm were divided into control and experimental groups. The experimental group was irradiated with a single absorbed dose of 2, 5, 10, and 15 Gy on the head and neck region. All the rats were sacrificed at 1, 3, 7, 14, 21, and 28 days after irradiation. The specimens including the submandibular gland were sectioned and observed using a immunohistochemical method. Results : In the 2 Gy group, clusterin expression was similar to that of the control group at 1 day after irradiation and it was observed in the striated ductal cells at 3 days after irradiation. In the 5 Gy group, clusterin expression was observed in the striated ductal cells at 1 day after irradiation and gradually increased in the 10 and 15 Gy groups. In the 15 Gy group, clusterin expression was prominent in the striated ductal cells at 1 day after irradiation, but it gradually decreased with the experimental period. The destruction of the striated ductal cells was observed in the 2 Gy group at 21 days after irradiation and in the 5, 10, and 15 Gy groups at 7 days after irradiation. The destruction of the acinar cells was observed in the 2 Gy group at 28 days after irradiation and in the 5, 10, and 15 Gy groups at 14 days after irradiation. Conclusion : Clusterin expression was induced by low doses of irradiation and it appeared to be involved in the regulation of cellular response to irradiation.