Journal of Oral Medicine and Pain

  • 제31권1호
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  • Pages.79-89
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  • 2006
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  • 2288-9272(pISSN)
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  • 2383-8493(eISSN)
대한안면통증구강내과학회 (Korean Academy of Orofacial Pain and Oral Medicine)
권호 표지

급만성 스트레스가 백서 악하선의 Clusterin 분비에 미치는 영향

Effects of Chronic and Acute Stress on Clusterin Secretion of the Rat Submandibular Gland

  • 진상배 (경희대학교 치과대학 구강내과학 교실) ;
  • 전양현 (경희대학교 치과대학 구강내과학 교실) ;
  • 홍정표 (경희대학교 치과대학 구강내과학 교실)
  • Jin, Sang-Bae (Department of Oral Medicine, College of Dentistry, Kyung-Hee University) ;
  • Chun, Yang-Hyun (Department of Oral Medicine, College of Dentistry, Kyung-Hee University) ;
  • Hong, Jung-Pyo (Department of Oral Medicine, College of Dentistry, Kyung-Hee University)
  • 발행 : 2006.03.30
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초록

구강 내에 발생되는 질환의 대부분이 타액의 영향을 받는다는 사실과 타액에 영향을 주는 전신적인 요소 중에서도 스트레스가 중요하다는 것은 이미 잘 알려져 있으나, 스트레스가 타액선에 미치는 영향에 관해서는 자율신경에 의한 거시적 반응에 대하여만 소개가 되었을 뿐 세포수준의 미시적 변화에 대하여는 별다른 언급이 없었다.. 이에 본 연구에서는 다양한 스트레스 조건하에서 백서의 악하선이 어떠한 변화를 보이는지를 clusterin 의 발현양상을 관찰함으로써 유추해보고자 하였다. 부여할 스트레스 조건을 급성 구속스트레스와 만성 저강도 스트레스의 두 가지로 정하고 7주된 Sprague-Dawley계 웅성백서 51마리를 사용하여 정해진 기간동안 급성 구속스트레스와 만성 저강도 비예측성 스트레스 (CUMS)를 가한 후 희생하여 악하선을 절취하고 역시 면역조직화학법과 웨스턴 면역점적법을 이용하여 악하선에서 clusterin발현의 시간에 따른 변화를 관찰하였으며, 그 결과는 다음과 같다: 1. 급성 구속스트레스 군에서, Clusterin 이 발현된 모든 선포의 합은 1시간 군을 제외한 모든 실험군 (9시간, 24시간, 72시간, 120시간, 그리고 168시간 군) 에서 대조군에 비하여 유의성 있게 감소하는 경향을 보였다 (p<0.001). 2. 만성저강도스트레스 군에서, 대조군에 대한 clusterin이 발현된 모든 선포의 합은, 2주군(p<0.01)에서 대조군에 비하여 유의하게 증가되는 모습을 보여주었고, 4주군(p<0.01)과 5주군(p<0.001)에서 대조군에 비하여 유의성 있게 감소하는 경향을 보였다. 3. 만성 저강도 스트레스 실험에서 4주째까지는 대조군과 실험군간의 당선호도 차이에 있어서 유의한 변화가 보이지 않았으나, 5주째에 유의성 있는 감소를 나타내었다(p<0.001). 4. 만성 저강도 스트레스 부여군은 5주째 대조군에 비하여 유의성 있는 체중변화(p<0.001)를 보여주었으나, 수분섭취량의 변화는 유의성 있는 상관관계를 보여주지 못하였다. 5. 면역점적검사를 시행한 결과, 구속스트레스 군에서는 clusterin의 발현이 시간에 따라 일정하게 감소하는 것으로 표현되었고, CUMS 군에서는 2주째까지는 증가하다가 3주 이후부터 실험 전기간에 걸쳐 감소하는 것으로 나타났으며 이는 당선호도 및 몸무게 변화의 양상의 변화와 크게 다르지 않았다. 따라서 위의 실험결과를 놓고 볼 때, 타액선 clusterin의 발현이 급성과 만성에 관계없이 스트레스 부여 후에 감소하였지만, 다른 문헌에서 제안된 것처럼 clusterin 의 고갈에 의한 세포자사적 변화는 관찰되지 않았다. 따라서 향후 보다 강화된 강도의 급성스트레스를 부여하는 방법과 더 장기적으로 진행된 연구를 통하여 고강도 스트레스에서 clusterin의 발현감소와 함께 세포의 변성이나 자사가 초래되는지, 저강도 스트레스의 경우 장기간 시간이 경과함에 따라 원래의 상태에 가깝게 회복되는지, 다른 열 충격 단백질이나 세포 사멸 시에 나타나는 단백질들을 동시에 확인하여 보는 것이 필요할 것으로 사료된다.

The aim of this study is to know how the rat submandibular gland changes under various emotional stress condition, using molecular biological methods. Restraint and chronic unpredictable mild stress (CUMS) experiment is conducted on fifty one 7-week old Sprague-Dawley rats (restraint stress experiment: 21, CUMS: 30). The rats were sacrificed, the submandibular glands were excised immediately at certain time, and examined by the use of immunohistochemistry and western blotting. In CUMS experiment, sucrose preference test, water intake change, weight change were implemented at 1 week interval for the experimental period The results are as follows: 1. The number of clusterin-secreting cells of restraint stress group compared to control group showed significantly decreasing tendency in all experimental groups except for the 1st hour group (p<0.001 in the 9th, 24th, 72nd, 120th, and 168th hour group). 2. The number of clusterin-secreting cells of CUMS group compared to control group showed significantly increasing tendency in the 2nd week group (p<0.01), and significantly decreasing tendency in the 4th and 5th week group (p<0.001). 3. Sucrose preference test in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01). 4. Weight change in CUMS experiment showed significant difference between the 5th week experimental group and control group (p<0.01), but water intake change didn't show significant difference compared to control group. 5. In western blot analysis, clusterin expression was decreased on a gradual basis in due time compared to the control group in the restraint stress group. As for CUMS group (chronic unpredictable mild stress group), it was increased till the 2nd week and decreased till the 5th week after that, which is similar to immunohistochemical analysis result and the decreasing tendency of sucrose preference and weigh changes. Through the test, it was proved that expression of clusterin in saliva glands decreases after receiving either acute or chronic stress, indicating relation with depression caused by chronic stress. Unlike other data, however, apoptotic tendency was hardly found in tissues. Diverse possibilities could be suggested on that: first, the stress was not enough to expedite apoptosis; second, apoptosis-related protein was already being secreted though not detected with microscope; third, clusterin, a major secretion molecule of saliva, decreased with saliva's malfunction due to stress. In the respect, it will be necessary to examine proteins expressed in case of cell death or other heat-shock proteins at the same time, in order to see whether any cellular change or death is caused by decreasing clusterin under high stress, and whether the original state is restored as time goes by under mild stress, through longer-term tests using even higher acute stress.

키워드

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