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

• Journal of Oral Medicine and Pain
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• v.33 no.3
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• pp.247-256
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• 2008

The belief that stress leads to illness has a long history. A number of the orofacial disease are also closely associated with stress. Despite research in the relationship of stress and the orofacial diseases leading to statistically significant correlations, the pathology remains vague. In the present study, the expression of clusterin, a stress protein responsible for antiapoptosis and cytoprotection, under restraint stress condition was observed in the submandibular gland, one of the major salivary glands. Sprague-Dawley rats were divided into 2 groups: normal group and restraint stress group. The rats of restraint stress group were placed in the stress cages and then sacrificed at day 0, 3 and 5 day of the experiment. After that, the submandibular glands of all the rats were excised immediately. The levels of clusterin proteins and mRNA in the tissues were measured by immunohistochemistry and Northern blot analyses, respectively. The results were as follows: 1. In the immunohistochemistry, clusterin protein was detected only immediately after the application of restraint stress. 2. In the restraint stress group, at day 3 and 5, histologically apoptosis was induced with karyorrhectic and pyknotic changes. 3. By the restraint stress, acinic cells were destructed earlier than ductal cells. 4. In the Northern blot, mRNA of clusterin was expressed only immediately after the application of restraint stress. The overall results suggest that as an early response to stress, clusterin is expressed in the glands to protect the glandular cells from the stress. But if stress is so strong and prolonged that it can exceed the stress adaptability of the cells, then the cells may undergo apoptosis instead of producing clusterin. An Epidemiologic Study of Symptoms of Temporomandibular Disorders in Korean College Students.

• The Journal of the Korean dental association
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• v.40 no.12 s.403
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• pp.917-927
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• 2002

• Environmental Analysis Health and Toxicology
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• v.23 no.4
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• pp.291-299
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• 2008

Bisphenol A diglycidyl ether(BADGE)는 비스페놀 A와 에피클로로하이드린의 축합에 의해 만들어지는 물질로 상업용 액상 에폭시 수지의 주성분이다. 본 연구는 clusterin mRNA 발현이 BADGE의 노출된 생식기계 독성에 연관되어 있는지를 연구하기 위해 수행하였다. BADGE는 SPF Sprague-Dawley 임신 랏트에 임신 6일부터 수유기가지 하루에 한 번 0(대조군)과 375mg/kg/day를 경구 투여하였다. 수컷 새끼는 일반 사항과 몸무게, 일반 발달 지표(예, 항문과 생식기 사이의 거리, 이개개전, 절치붕출, 유두잔류, 안검개열, 고환하강, 포피박리 등)등을 관찰하였다. 대조군과 투여군에서 다섯 마리의 수컷 새끼는 출생 후 3, 6와 9주에 부검하여 부고환의 조직학적 변화 등을 관찰하였다. BADGE 375 mg/kg/day 투여군에서 항문과 생식기 사이의 거리는 대조군보다 길어지는 경향을 보였다. 출생 후 6주와 9주에서 부고환의 상대 무게는 대조군보다 약간 증가하였으나 조직학적인 변화는 관찰되지 않았다. BADGE 투여 군에서 clusterin mRNA 발현량은 대조군에 비해 3주에 56%, 6주에 57% 그리고 9주에 86% 감소하였다. 이런 결과는 랏트의 부고환에서 clusterin은 BADGE에 반응하는 유전자 중 하나일 수 있다는 가능성을 나타낸다.

• Journal of Oral Medicine and Pain
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• v.25 no.3
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• pp.279-289
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• 2000

현대에는 날로 복잡해지는 생활양식의 변화에 따라 다양한 스트레스에 노출되고 그로 인한 면역계, 신경계 또는 각종 장기의 기능이상이 점차로 증가되고 있다. 최근 치과에 내원하는 환자들 중 구강건조증을 호소하는 환자들의 다수는 스트레스에 노출되어 있고, 임상적으로도 방사선 타액선 기능 검사에서 타액선, 특히 악하선의 기능이 현저하게 저하되어 있는 소견을 관찰할 수 있 다. 더욱이 이는 타액선 조영술에서 이미 보고된 바 있는 면역성 질환인 $Sj\ddot{o}gren$ 증후군과는 다른 양상으로 관찰되고 있어, 이에 본인은 백서에 스트레스와 관련이 깊은 내분비적 환경 변화를 유도함으로써 이와같은 변화가 악하선 조직의 병리적 변화와 어떠한 연관성을 갖는 지를 관찰하고자 본 실험을 시행하였다. 생후 7주된 Sprague-Dawley계 웅성 백서(165-209g/bw) 40마리를 2 개의 실험군(부신 제거군 : ADX 군, 부신 제거 후 dexamethasone 투여군 : DEX 군)으로 나누어 실험하였다. ADX 군은 외과적으로 부신을 제거한 후 다른 조건을 부여하지 않았고, DEX 군은 외과적으로 부신을 제거한 후 매일 dexamethasone($1.5*10^{-46}mg/g$ I.V./day)을 투여하였으며, 이들을 각각 실험 후 즉시, 1일, 3일, 5일, 7일에 희생시켰다. 그 후에는 즉시 악하선을 적출하여 면역조직화학법으로 Clusterin의 발현 정도 및 부위를 관찰하였다. 그 결과는 다음과 같았다. 1. 광학현미경 하에서, 양 군 모두 유의할만한 조직학적 변화는 관찰되지 않았다. 2. ADX 군에서는 실험기간 전반에 걸쳐 도관세포와 선포세포에서 clusterin이 발현되었다. 3. DEX 군에서는 소수의 선세포에서 국소적으로 clusterin이 관찰되었으나, 전반적으로 도관세포, 선포세포에서 공히 clusterin이 관찰되지 않았다. 부신을 제거한 군에서는 실험기간 전반에 걸쳐 clusterin이 발현되었는데, 이는 clusterin이 스트레스의 생리적 반응의 결과로서 세포보호작용을 한다는 사실에 기초하여 볼 때, 부신을 제거하였을 때도 스트레스를 받았을 때와 같은 영향이 백서의 악하선에 가해졌을 것으로 생각된다. 반면, 부신제거후 glucocorticoid agonist인 dexamethasone을 투여하였을 때, clusterin이 감소 내지 관찰되지 않았던 것은 부신제거에 의해 악하선에 가해졌던 영향을 dexamethasone이 길항하여 나타난 결과로 볼 수 있어, 스트레스에 의해 부신으로부터 분비되는 glucocorticoid가 타액선을 보호하는데 중요한 작용을 하는 것을 시사한다.

• Journal of Oral Medicine and Pain
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• v.24 no.3
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• pp.245-257
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• 1999

일상 생활에서 우리는 스트레스에 항상 노출되어 있으며, 스트레스는 생체의 신경계, 내분비계 및 면역계의 변화를 수반한 항상성의 파괴로 수많은 정신적, 육체적 질병을 야기시킨다. 특히 구강안면영역에서도 다양한 구강점막질환과 구강건조증 등을 발생시킨다. 스트레스를 제거하는 방법으로는 약물요법 및 상담, 명상요법, 종교요법 등 다양한 방법이 제시되고 있는데, 다소의 부작용이 나타나거나 꾸준히 시행하기가 쉽지 않으며 스트레스의 원인을 근본적으로 제거하기가 현실적으로 용이하지 않은 경우가 많아 스트레스에 대한 해결책에 대하여 많은 관심이 집중되고 있다. 이에 본인은 스트레스가 가해졌을 때 백서 악하선에서 관철되며 apoptosis에 대하여 세포보호작용을 하는 clusterin(SGP-2)을 이용하여 구속스트레스를 가하기에 앞서 오랫동안 경험적으로 사용되어 왔고 부작용이 적은 전통약물인 보혈안신탕을 투여하고 스트레스에 의한 타액선의 조직변화를 관찰하여 그 효과를 확인해 보고자하였다. Sprague-Dawley계 응성 백서(200-230g/bw) 33마리를 정상 대조군(3마리), 구속스트레스군(15마리) 및 보혈안신탕 투여 후 구속스트레스군(15마리)으로 나누고 이틀을 각각 구속장치에 구속한 후 0, 1, 3, 5, 7일에 회생시켜 악하선을 적출하였으며, 면역조직화학법 및 Northern Blot을 이용하여 clusterin의 변화를 관찰하였다. 그 결과는 다음과 같다. 1. 구속스트레스군의 악하선 조직에서 clusterin 단백질과 mRNA는 실험 즉일군에서만 미약하게 관찰되었으며 실험 3일과 5일 후에 핵붕괴 및 핵농축 등의 핵변화를 동반한 apoptosis가 관찰 되었다. 2. 보혈안신탕 투여 후 구속스트레스군의 악하선 조직에서 실험 5일군까지 clusterin 이 증가한 후 실험 7일군에서는 감소하였다. 3. 보혈안신탕 투여 후 구속스트레스군의 악하선 조직에서는 apoptosis가 관찰되지 않았다. 4. 보혈안신탕 투여 후 구속스트레스군의 악하선 조직에서 clusterin mRNA가 실험 전군에 걸쳐 미약하게 관찰되었다. 이상의 결과로 타액선 조직은 스트레스 단백질인 clusterin을 생산하여 세포를 보호함으로써 스트레스 상황에 적응하지만, 생리적 적용한계를 넘는 스트레스에 노출될 때는 apoptosis됨이 확인되었다. 그리고 보혈안신탕은 스트레스 상황에서 세포의 생리적 적응력을 높여 세포의 apoptosis를 억제하는 효과를 나타냄이 확인되었다. 따라서 본 연구결과는 구강건조증등의 스트레스성 타액선 질환의 병리기전을 규명하는데 도움이되리라 생각되며, 향후 항스트레스 효과를 가진 보혈안신탕등의 한약재를 임상에 적용함으로써 스트레스로 인한 신체의 병리적 변화를 다소나마 차단할 수 있을 것으로 사료된다.

• The Korean Journal of Physiology and Pharmacology
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• v.28 no.3
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• pp.219-227
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• 2024

Bladder cancer remains the 10th most common cancer worldwide. In recent years, metformin has been found to have potential anti-bladder cancer activity while high concentration of IC₅₀ at millimolar level is needed, which could not be reached by regular oral administration route. Thus, higher efficient agent is urgently demanded for clinically treating bladder cancer. Here, by conjugating artesunate to metformin, a novel artesunate-metformin dimer triazine derivative AM2 was designed and synthesized. The inhibitory effect of AM2 on bladder cancer cell line T24 and the mechanism underlying was determined. Anti-tumor activity of AM2 was assessed by MTT, cloning formation and wound healing assays. Decreasing effect of AM2 on lipogenesis was determined by oil red O staining. The protein expressions of Clusterin, SREBP1 and FASN in T24 cells were evaluated by Western blotting. The results show that AM2 significantly inhibited cell proliferation and migration at micromolar level, much higher than parental metformin. AM2 reduced lipogenesis and down-regulated the expressions of Clusterin, SREBP1 and FASN. These results suggest that AM2 inhibits the growth of bladder cancer cells T24 by inhibiting cellular lipogenesis associated with the Clusterin/SREBP1/FASN signaling pathway.

• Molecules and Cells
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• v.20 no.1
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• pp.97-104
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• 2005

To investigate apoptosis in HC11 mammary epithelial cells, we compared the gene expression profiles of actively growing and serum-starved apoptotic cells using a mouse apoptosis gene array and $^{33}P$-labeled cDNA prepared from the RNA of the two cultures. Analysis of the arrays showed that expression of several genes such as clusterin, secreted frizzled related protein mRNA (sFRP-1), CREB-binding protein (CBP), and others was higher in the apoptotic cells whereas expression of certain genes including survivin, cell division cycle 2 homolog A (CDC2), and cyclin A was lower. These expression patterns were confirmed by RT-PCR and/or Northern analyses. We compared the expression of some of these genes in the mouse mammary gland under various physiological conditions. The expression levels of genes (clusterin, CBP, and M6P-R) up-regulated in apoptotic conditions were higher at involution than during lactation. On the other hand, genes (Pin, CDC2) downregulated in apoptotic conditions were relatively highly expressed in virgin and pregnant mice. We conclude that certain genes such as clusterin, sFRP-1, GAS1 and CBP are induced in apoptotic mammary epithelial cells, and others are repressed. Moreover, the apoptosis array is an efficient technique for comparing gene expression profiles in different states of the same cell type.

• Journal of Oral Medicine and Pain
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• v.38 no.2
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• pp.121-136
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• 2013

It has been known that saliva may affect the most of oral diseases. On the contrary, several systemic conditions may affect salivary flow and cause oral dryness and psychosocial stress especially may a crucial role in the etiology of hyposalivation and oral dryness. Many studies have focused on macroscopic effects of the stress on the salivary glands by autonomic response, but on the other hand it has hardly been reported on cellular microscopic effects of the stress on the salivary glands. Therefore, this study was performed to examine clusterin, a antiapoptotic and cytoprotective protein, in the parotid glands under restraint stress condition. 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 7 days after application of the stress and the both parotid glands were excised. Immunohistochemistry and electron microscopy were performed. The finding were as follows: 1. In parotid glands, repeated stress denaturalize the acinar cells, interacinous tissues and interacinous connective tissues were separated to individual acinar cells. After 4 days of experiment, there were lots of vacuoles and intercalated ducts. 2. In parotid glands, repeated stress make the rER which is in acinar cells swollen after 3 days of experiment and it was intensified to 4 days. After 5 days of experiment the edema got worse and degenerated. 3. In parotid glands, clusterin was reduced in ductal cell cytoplasm but in intercalated duct clusterin was slightly stained until 3 days prominently increased until 4 days and then decreased again after 5 days of experiment.

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