Barthod F
Service de chirurgie générale, digestive et vasculaire, Hôpital A. Paré, Boulogne sur Seine.
J Chir (Paris). 1994 May;131(5):221-35.
Ischaemia of the small intestine leads to the destruction of the intestinal mucosa. The capacity of the epithelium to regenerate is proportional to the duration of revascularization. The aim of this work was to analyze the kinetic aspects of intestinal epithelial regeneration after destruction due to prolonged ischaemia. This study was conducted in 44 animals (swine) after development of an ischaemia-revascularization protocol of a jejunal loop and bipolar secondary cutaneous exteriorization. After a first series with ischaemia times of 1, 2, 3 and 4 hours, the 4 hour period of ischaemia was chosen for further analysis of the regeneration kinetics over a period of 21 days since it leads to regular and total destruction of the epithelium compatible with regeneration. This analysis included (1) a histological examination (semi-thin slices), (2) immunofluorescent detection of intestinal brush border proteins on frozen slices (villin, saccharase-isomaltase, aminopeptidase N, dipeptidylpeptidase-IV) and mucines, (3) measurement of specific intestinal hydrolase activities (saccharase, aminopeptidase N, dipeptidylpeptidase-IV and alkaline phosphatase) in enriched brush border fractions, and (4) an analysis of variations in intestinal flora. After the 4 hour ischaemia, total destruction of the epithelium with disappearance of the villin and intestinal hydrolases and disorganization of the mucosa invaded by mucosal lacks was observed. Epithelial regeneration was rapid and two days later the histological aspect of the mucosa showed apical expression (still discontinuous), villin and intestinal hydrolase activity. Luminal apical expression of the markers became continuous on day 4, demonstrating the total recovery of the intestinal barrier as confirmed by stable microbial flora. Mucine expression also returned to normal. This regeneration was however incomplete since the mucosa was seen to be flat, without villosities. Immunofluorescence showed the weak intensity of brush border activity and the very low specific activity of hydrolase. Values were below normal and did not start to rise again until day 21. If serum levels and associated brush border markers could be measured and were significant, they could be specific markers of regeneration in double stomy ischaemic-revascularized intestine and thus eliminate the need for early second look laparotomy.
小肠缺血会导致肠黏膜破坏。上皮细胞的再生能力与血管再通的持续时间成正比。本研究的目的是分析长时间缺血导致破坏后肠上皮再生的动力学方面。本研究在44只动物(猪)身上进行,采用空肠袢缺血-再灌注方案及双极二期皮肤外置术。在进行了缺血时间为1、2、3和4小时的第一组实验后,选择4小时缺血期对21天内的再生动力学进行进一步分析,因为该时长会导致与再生相适应的上皮细胞规律性且完全的破坏。该分析包括:(1)组织学检查(半薄切片);(2)在冰冻切片上对肠刷状缘蛋白(绒毛蛋白、蔗糖酶-异麦芽糖酶、氨肽酶N、二肽基肽酶-IV)和黏蛋白进行免疫荧光检测;(3)在富集的刷状缘组分中测量特定肠水解酶活性(蔗糖酶、氨肽酶N、二肽基肽酶-IV和碱性磷酸酶);(4)分析肠道菌群的变化。4小时缺血后,观察到上皮细胞完全破坏,绒毛蛋白和肠水解酶消失,黏膜被黏膜缺损侵袭而紊乱。上皮再生迅速,两天后黏膜的组织学表现显示顶端表达(仍不连续)、绒毛蛋白和肠水解酶活性。标志物的管腔顶端表达在第4天变得连续,表明肠屏障完全恢复,稳定的微生物菌群也证实了这一点。黏蛋白表达也恢复正常。然而,这种再生并不完全,因为黏膜看起来是扁平的,没有绒毛。免疫荧光显示刷状缘活性强度较弱,水解酶的比活性非常低。数值低于正常水平,直到第21天才开始再次上升。如果血清水平及相关刷状缘标志物能够被测量且具有显著性,它们可能成为双腔造口缺血-再灌注肠段再生的特异性标志物,从而无需早期再次剖腹探查。