نتيجه ي رنگ آميزي ايمونوهيستوشيمي در تعيين اختلال عملكرد روده بعد از عمل در كودكان مبتلا به بيماري هيرشپرونگ

The Immunohistochemical Staining in Determination of Postoperative Bowel Dysfunction in Children with Hirschsprung's Disease

ﻣﻘﺪﻣﻪ: اﯾﻦ ﻣﻄﺎﻟﻌﻪ، ﺑﺎ ﻫﺪف ارزﯾﺎﺑﯽ رﻧﮓ آﻣﯿﺰي ﮐﺎﻟﺮﺗﯿﻨﯿﻦ و ﻧﯿﺘﺮﯾﮏ اﮐﺴﯿﺪ در ﺑﺨﺶ ﮔﺎﻧﮕﻠﯿﻮﻧﯿﮏ روده ي ﺑﺰرگ ﺑﯿﻤﺎران ﻫﯿﺮﺷﭙﺮوﻧﮓ )Hirschsprung's disease ﯾﺎ HD( اﻧﺠﺎم ﺷﺪ. روش ﻫﺎ: در ﯾﮏ ﻣﻄﺎﻟﻌﻪ ي ﻣﻘﻄﻌﯽ،60 ﮐﻮدك ﻣﺒﺘﻼ ﺑﻪ ﻫﯿﺮﺷﭙﺮوﻧﮓ وارد ﻣﻄﺎﻟﻌﻪ و ﺑﺎ ﻋﻤﻞ ﺟﺮاﺣﯽ، روده از ﻣﻘﻌﺪ ﺑﯿﺮون ﮐﺸﯿﺪه و ﻫﺮ 5 ﺳﺎﻧﺘﯽ ﻣﺘﺮ از آن ﺑﯿﻮﭘﺴﯽ ﺷﺪ و ﺑﺨﺶ ﭘﺮوﮔﺰﯾﻤﺎل ﻧﻤﻮﻧﻪ ﺑﺮداري ﺷﺪه، ﺑﺎ رﻧﮓ آﻣﯿﺰي ﻫﻤﺎﺗﻮﮐﺴﯿﻠﯿﻦ- اﺋﻮزﯾﻦ )H&E( و اﯾﻤﻮﻧﻮﻫﯿﺴﺘﻮﺷﯿﻤﯽ )IHC( ﺑﺮرﺳﯽ ﺷﺪ و ﻧﺘﯿﺠﻪ ﺑﺎ ﻣﺜﺒﺖ ﺿﻌﯿﻒ ﯾﺎ ﻣﺜﺒﺖ ﺷﺪﯾﺪ ﺑﺮاي ﮐﺎﻟﺮﺗﯿﻨﯿﻦ و ﻧﯿﺘﺮﯾﮏ اﮐﺴﯿﺪ ﮔﺰارش ﺷﺪ. ﮐﻠﯿﻪ ي ﺑﯿﻤﺎران در 1 و 6 ﻣﺎه ﺑﻌﺪ از ﻋﻤﻞ ﭘﯽ ﮔﯿﺮي ﺷﺪﻧﺪ و ﻧﺘﯿﺠﻪ ي ﻋﻤﻞ در دو ﮔﺮوه ﻣﻘﺎﯾﺴﻪ ﮔﺮدﯾﺪ. ﯾﺎﻓﺘﻪ ﻫﺎ: از 60 ﮐﻮدك ﻣﺒﺘﻼ ﺑﻪ ﻫﯿﺮﺷﭙﺮوﻧﮓ، 14 ﻧﻔﺮ )23/3 درﺻﺪ( ﻋﻼﻣﺖ دار ﺑﻮدﻧﺪ. ﻫﻤﻪ ي ﻧﻤﻮﻧﻪ ﻫﺎي ﺑﺨﺶ ﻫﺎي ﮔﺎﻧﮕﻠﯿﻮﻧﯽ ﮐﻮﻟﻮرﮐﺘﺎل ﺑﯿﻤﺎران ﻋﻼﻣﺖ دار، از ﻧﻈﺮ رﻧﮓ آﻣﯿﺰي ﻧﯿﺘﺮﯾﮏ اﮐﺴﯿﺪ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ رﻧﮓ آﻣﯿﺰي ﻧﯿﺘﺮﯾﮏ اﮐﺴﯿﺪ ﺟﻔﺖ، ﻣﻨﻔﯽ ﺑﻮدﻧﺪ. ﺑﺮرﺳﯽ اﯾﻤﻮﻧﻮﻫﯿﺴﺘﻮﺷﯿﻤﯽ، رﻧﮓ آﻣﯿﺰي ﻫﺴﺘﻪ اي و ﺳﯿﺘﻮﭘﻼﺳﻤﯽ ﮐﺎﻟﺮﺗﯿﻨﯿﻦ ﻧﺸﺎن داد ﮐﻪ ﺳﻠﻮل ﻫﺎي ﮔﺎﻧﮕﻠﯿﻮﻧﯽ در 52 ﺑﯿﻤﺎر ﺷﺪﯾﺪا ﻣﺜﺒﺖ و در 8 ﺑﯿﻤﺎر ﻣﺜﺒﺖ ﺿﻌﯿﻒ ﺑﻮد. 6 ﺑﯿﻤﺎر در ﮔﺮوه ﺷﺪﯾﺪا ﻣﺜﺒﺖ )11/5 درﺻﺪ( ﻋﻼﻣﺖ دار ﺑﻮدﻧﺪ؛ در ﺣﺎﻟﯽ ﮐﻪ در 8 ﺑﯿﻤﺎر ﮐﻪ رﻧﮓ آﻣﯿﺰي ﮐﺎﻟﺮﺗﯿﻨﯿﻦ ﻣﺜﺒﺖ ﺿﻌﯿﻒ ﮔﺰارش ﺷﺪه ﺑﻮد، ﻫﻤﻪ ي ﻋﻼﯾﻢ وﺟﻮد داﺷﺖ )0/001 = P(. دﻗﺖ ﮐﻠﯽ ﭘﯿﺶ ﺑﯿﻨﯽ ﯾﺒﻮﺳﺖ ﺑﻌﺪ از ﻋﻤﻞ ﺑﺮاي رﻧﮓ آﻣﯿﺰي ﮐﺎﻟﺮﺗﯿﻨﯿﻦ 96/67 درﺻﺪ ﺑﻮد. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﯾﺎﻓﺘﻪ ﻫﺎي ﻣﻄﺎﻟﻌﻪ ي ﺣﺎﺿﺮ، ﺑﻪ ﻧﻈﺮ ﻣﯽ رﺳﺪ در ﮐﻮدﮐﺎن ﻣﺒﺘﻼ ﺑﻪ ﻫﯿﺮﺷﭙﺮوﻧﮓ ﺗﺤﺖ ﻋﻤﻞ ﺟﺮاﺣﯽ، رﻧﮓ آﻣﯿﺰي اﯾﻤﻮﻧﻮﻫﯿﺴﺘﻮﺷﯿﻤﯽ ﭘﺮوﮔﺰﯾﻤﺎل ﻗﺴﻤﺖ ﮔﺎﻧﮕﻠﯿﻮﻧﯽ ﺑﺮاي ﺗﻌﯿﯿﻦ اﺧﺘﻼل ﻋﻤﻠﮑﺮد روده ﺑﻌﺪ از ﻋﻤﻞ، ﻣﯽ ﺗﻮاﻧﺪ ﻣﺆﺛﺮ ﺑﺎﺷﺪ.

Background: The aim of this study was to evaluate the calretinin and nitric oxide (NO) immunohistochemical (IHC) staining in ganglionic segment of colon of children with Hirschsprung's disease (HD). Methods: In a cross-sectional study, 60 children with HD entered the study, and underwent surgery to pull out the bowel from the anus to take biopsy from every 5 cm. Hematoxylin and eosin (H&E) and IHC methods were evaluated, and the results were reported to be poorly positive or severely positive for calretinin. All patients were followed up 1 and 6 months after the operation, and the results were compared between the two groups. Findings: From 60 children with HD, 14 patients (23.3%) were symptomatic. In 14 symptomatic patients, all colorectal samples from ganglionic segments were negative for NO staining in comparison of placental NO staining as sham group. In the IHC examination, nuclear and cytoplasmic calretinin staining revealed ganglion cells in 52 patients as strongly positive, and weakly positive in 8 patients. Six patients in strongly positive group (11.5%) were symptomatic, while 8 patients whose calretinin staining was reported as weakly stained, had symptoms (P = 0.001). Overall accuracy of prediction of postoperative constipation for calretinin staining was 96.67%. Conclusion: According to the findings of our study, it seems that in children with HD who undergo surgery, proximal ganglion immunohistochemical staining may be effective in determining postoperative bowel dysfunction.