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AN IMMUNOHISTOCHEMICAL STUDY ON DNA SYNTHESIS OF SALIVARY GLAND TISSUE CELLS AND ENDOTHELIAL CELL AFTER IRRADIATION
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KMID : 0378219910210020183
Abstract
ÀúÀÚ´Â 150gm ¿õ¼º ¹é¼ 40¸¶¸®ÀÇ ¾ÇÇϼ±¿¡ 2Gy, 5Gy, l0GyÀÇ ¹æ»ç¼±À» °¢°¢ ´ÜÀÏÁ¶»ç
ÈÄ 1½Ã°£, 3½Ã°£, 7½Ã°£, 15½Ã°£, 1ÀÏ, 3ÀÏ, 5ÀÏ, 7ÀÏ ÈÄ¿¡ bromodeoxyuridine(Sigma)À» 0.1§·
/gÀ» º¹°³» ÁÖ»ç 2½Ã°£ ÈÄ¿¡ Èñ»ý½ÃÅ°°í H&E, PAS¿°»öÀ» ÇÏ¿© Ÿ¾×¼± Á¶Á÷¼¼Æ÷ÀÇ º¯È
¸¦ °Ë°æÇÏ°í, Á¶»ç·®¿¡ µû¶ó Ÿ¾×¼± ¼¼Æ÷ ¹× Ç÷°ü ³»ÇǼ¼Æ÷ÀÇ DNAÇÕ¼º ºÐÆ÷¸¦ Á¶»ç Çϱâ
À§ÇÏ¿© Feulgen¿°»öÀ» ÇÏ°í, DNAÇÕ¼ºÀÇ º¯È¿¡ ´ëÇÑ ¿¬±¸¸¦ À§ÇÏ¿© Ç×-bromode-
oxyuridine(Sanbyo Co.)À» ÀÌ¿ëÇÏ¿© ¸é¿ªÁ¶Á÷ÈÇÐÀû ¹æ¹ýÀ» ½ÃÇàÇÏ¿´°í Á¶Á÷Ç¥º»¿¡¼ Çѽ½
¶óÀ̵å´ç 200¹è ºñÀ²·Î ÀÓÀÇ·Î 5±ºµ¥¸¦ Á¤ÇÏ¿© Ç×-BrdU¿¡ ¾ç¼ºÀΠŸ¾×¼±¼¼Æ÷¸¦ ±¤ÇÐ Çö¹Ì
°æÀ» »ç¿ëÇÏ¿© ¼ö¸¦ ¼¼¾î ´ÙÀ½°ú °°Àº °á°ú¸¦ ¾ò¾ú´Ù.
1. PAS¿°»ö °á°ú 5GyÁ¶»ç ÈÄ 3ÀÏ° Àå¾×¼º¼±¼¼Æ÷ÀÇ ºÐºñ°¨¼Ò°¡ ³ªÅ¸³µ°í l0GyÁ¶»ç ÈÄ 7
ÀÏ°¿¡´Â Á¡¾×¼º ¼±¼¼Æ÷ÀÇ ºÐºñ°¨¼Ò°¡ º¸¿´´Ù.
2. º´¸®Á¶Á÷ÇÐÀû ¼Ò°ß¿¡¼´Â Àå¾×¼º ¼±¼¼Æ÷ÀÇ º¯¼ºÀÌ 2Gy Á¶»ç ÈÄ 3ÀϺÎÅÍ °üÂûµÇ¾ú°í
Á¡¾×¼º ¼±¼¼Æ÷¿Í ºÐºñµµ°ü¼¼Æ÷´Â °ÅÀÇ º¯È°¡ ¾ø¾ú´Ù.
3. Feulgen¿°»ö¿¡¼ 2Gy, 5GyÁ¶»ç 3ÀÏ°ÀÌÈÄ¿¡´Â BrdU¿°»ö¿¡¼ º¸´Ù °³Àçµµ°ü¼¼Æ÷, Ⱦ¹®
µµ°ü¼¼Æ÷, ºÐºñµµ°ü¼¼Æ÷¿¡¼ DNAÇÕ¼ºÀÌ È°¹ßÇÏ¿´´Ù.
4. ¸é¿ªÁ¶Á÷ÈÇÐÀû ¼Ò°ß¿¡¼ Á¶»çÈÄ Ãʱ⿡ Àå¾×¼º ¼±¼¼Æ÷¿Í °ú¸³¼º À±°ü¼¼Æ÷¿¡¼ DNA
ÇÕ¼ºÀÌ ÇöÀúÈ÷ °¨¼ÒÇßÀ¸¸ç Á¶»çÈÄ 7ÀÏ°¿¡µµ DNAÇÕ¼ºÀÌ È°¹ßÇÏÁö ¾Ê¾ÒÀ¸³ª ºñ±³Àû S-±â
¿¡ ¼ÓÇÏ´Â ¼¼Æ÷°¡ ÀûÀº ºÐºñµµ°ü¼¼Æ÷¿Í Ⱦ¹®µµ °ü¼¼Æ÷´Â 2Gy, 5Gy·Î ¹æ»ç¼± Á¶»çÈÄ 7ÀÏ°
¿¡DNAÇÕ¼ºÀÌ Áõ°¡ÇÏ¿´´Ù.
5. ¸é¿ªÁ¶Á÷ÈÇÐÀû ¼Ò°ß¿¡¼ ¹æ»ç¼± Á¶»çÈÄ Ç÷°ü ³»ÇǼ¼Æ÷ÀÇ DNAÇÕ¼ºÀÌ °è¼Ó °¨¼ÒÇÏ¿´
À¸³ª 2Gy¿Í 5GyÁ¶»çÈÄ 7ÀÏ°¿¡´Â DNAÇÕ¼ºÀÌ Áõ°¡ÇÏ¿´´Ù.
#ÃÊ·Ï#
After single fraction of 2, 5, 10 Gy irradiation on submandibular gland of 40 male rats,
weighing 150gm, respectively, these animal were sacrificed two hours after 0.1 §·/g
bromodeoxyuridine (Sigma) peritoneal injection in 1, 3, 7, 15 hours, 1, 3, 7 days after
irradiation. And excised submandibular gland were fixed in Carnoy's and Bouin's
solution for 2 hours. Paraffin sections were stained with H&E, and PAS for the
observation of the change of salivary gland tissue, and with Feulgen for the study of
the DNA distribution, and immunohistochemically stained with anti-bromodeoxyuridine
(Sanbyo Co.) for detection of DNA synthetic cells in order to Study the distribution of
DNA synthetic cells of salivary gland tissue and endothelium after irradiation in 5
different sites of 6 slides on X 200 high power field.
The results were as followings.
1. In PAS staining 3 days after 5Gy irradiation, decreased mucine secretion of serous
cellswere found, and 7 days after l0Gy irradiation, decreased mucine secretion of mucous
cells were found.
2. In histopathologic features, degeneration of serous cells were found in 3 days after
2 Gy irradiation and there was little change in mucous cells and excretory duct cells.
3. In Feugen staining, 3 days after 2 Gy, 5 Gy irradiation, more high percentage of
DNA syn-thetic cells were found in intercalated duct cells, striated duct cells and
excretory duct cells than in BrdU staining.
4. In immunohistochemical features, DNA synethsis of serous cells and granular
convoluted tubular cells abruptly decreased in early period after irradiation and showed
no recovery in 7 days after irradiation but there was an increase in DNA synthesis of
intercalated duct cells, striated duct cells and excretory duct cells, which have less
S-phase cells comparatively, in 7 days after 2 Gy, 5 Gy irradiation.
5. In immunohistochemical features, the DNA synthesis of endothelial cells was
continuously decreased after irradiation but showed slight increase in 7 days after 2 Gy
and 5 Gy irradiation.
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