AgNO_3溶液对兔肠去粘膜作用的实验研究
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摘要
目的：观察不同浓度的AgNO_3溶液在不同时间内对兔肠的去粘膜作用，探讨一种新的去粘膜方法。
方法：将40只大白兔随机分4组，每组10只，手术游离15cm空肠，保留该肠段血供，肠吻合恢复肠的连续性。除A组外，对其余三组(B、C、D组)游离肠段分别给于2％ AgNO_3溶液30min、5％ AgNO_3溶液10min、手术剥除肠粘膜处理，分别测量各组游离肠段在内压为14cmH_2O时的容量(是正常兔膀胱在排尿瞬间时的充盈压平均值，游离肠段在内压14cmH_2O时的容量可以间接反映肠膀胱充盈状态下肠壁的顺应性)。各组于第8周末处死取游离空肠，测定其容量，制备HE染色、PAS染色及masson's trichrome(马松三色)染色切片，进行病理组织学检查及分析。
结果：8周末HE染色显示：A组肠粘膜完整。B组肠粘膜、粘膜肌层部分缺损，部分再生粘膜下无粘膜肌。C组肠粘膜完全缺失，肠壁表层覆盖着一层由大量纤维素、炎性细胞、坏死组织构成的假膜。D组无粘膜层，粘膜下层由瘢痕组织替代。PAS染色显示：A组全部阳性，B组部分阳性，C组、D组无阳性染色。马松三色染色显示：A组、B组、C组肠粘膜下层无明显胶原纤维增厚，D组有明显胶原纤维增厚。胶原纤维沉淀指数同A组比较，B、C组术后无明显变化，P＞0.05；D组明显增厚，P＜0.01。各组术中和术后8周游离肠容量的比较：A、B、C组无显著差异，P＞0.05；D组差异有显著性，P＜0.01。
结论：AgNO_3溶液有去肠粘膜作用，5％AgNO_3溶液10min处理肠段可以完全去除肠粘膜，无显著纤维化，无显著肠管挛缩，对肠膀胱成形术具有重要的临床意义。
Objective: To study the de-epithelialization effect of different concentrations of AgNO3 solutions at different time points on intestines of rabbits and explore its clinical significance on enterocystoplasy.
Methods: Forty rabbits were randomly divided into four groups (10 rabbits in each group). For each rabbit, one 15 cm segment of jejunum was isolated with intact mesentety. Intestinal anastomosis was performed to keep its continuity. The segments in group B were treated with 2g/100ml silver nitrate (AgNO3) solution for 30 minutes, while the segments in group C were treated with 5g/100ml AgNO3 solution for 10 minutes,and the mucosa of the segments in group D were stripped by surgical blade. The The segments in Group A were only treated with nomal Saline. Volume of free intestine was measured when the inner pressure was 14cmH2O(The pressure is the average pressure in normal rabbits bladder at the moment of miction. And the volume of free intestine at this pressure could reflect indirectly the compliance of the intestinal wall of the substitute bladder.). Thirty-three rabbits survived and were killed 8 weeks after operation, Volume of free intestines were measured too, and those free intestines were produced to
sections stained with HE, PAS and masson's trichrome for histopathological analysis by light microscope.
Results: At the end of the 8th week, it was displayed by HE stain that the intestinal mucosa layer in the samples of group A was integrated. In the samples of group B, there was defect in some parts of the intestinal mucosa layer and muscularis mucosa layer, and there weren't any muscularis mucosa below some parts of regeneration mucosa. In the samples of group C, there was a totally depletion of mucosa and there was a layer of false membrane constituted of cellulose, inflammatory cells and necrosis tissues over the intestinal wall. In the samples of group D, there was no mucosa layer and the submucosa layer was substituted by scar tissues. It was showed in PAS stain that there were all positive stains in group A, part of positive stains in group B and completely negative stains in group C and D. It was showed in masson's trichrome stain that there was no significant thickening of the collagen fiber in the submucosa layer in group A, B and C, and apparently thickening in group D. Comparing with group A, the
collagen fiber despot index in group B and group C had no significant change after the operation (P>0.05). But there was significant change in group D (P<0.01). And in groups A, B and C, there were no significant difference between the volumes of free
intestines during the operation and those at the 8th week after the operation (P>0.05). But in group D there were significant difference between the intestinal volumes during the operation and those at the 8th week after the operation (P<0.01).
Conclusion: AgNO3 solution can effectively inactivate intestinal mucous. Ten minutes' treatment of 5% AgNO3 solution can successfully de-epithclialize jijunal segments without creating severe retraction or fibrosis. This novel treatment can be very useful for enterocystoplasy.
方法：将40只大白兔随机分4组，每组10只，手术游离15cm空肠，保留该肠段血供，肠吻合恢复肠的连续性。除A组外，对其余三组(B、C、D组)游离肠段分别给于2％ AgNO_3溶液30min、5％ AgNO_3溶液10min、手术剥除肠粘膜处理，分别测量各组游离肠段在内压为14cmH_2O时的容量(是正常兔膀胱在排尿瞬间时的充盈压平均值，游离肠段在内压14cmH_2O时的容量可以间接反映肠膀胱充盈状态下肠壁的顺应性)。各组于第8周末处死取游离空肠，测定其容量，制备HE染色、PAS染色及masson's trichrome(马松三色)染色切片，进行病理组织学检查及分析。
结果：8周末HE染色显示：A组肠粘膜完整。B组肠粘膜、粘膜肌层部分缺损，部分再生粘膜下无粘膜肌。C组肠粘膜完全缺失，肠壁表层覆盖着一层由大量纤维素、炎性细胞、坏死组织构成的假膜。D组无粘膜层，粘膜下层由瘢痕组织替代。PAS染色显示：A组全部阳性，B组部分阳性，C组、D组无阳性染色。马松三色染色显示：A组、B组、C组肠粘膜下层无明显胶原纤维增厚，D组有明显胶原纤维增厚。胶原纤维沉淀指数同A组比较，B、C组术后无明显变化，P＞0.05；D组明显增厚，P＜0.01。各组术中和术后8周游离肠容量的比较：A、B、C组无显著差异，P＞0.05；D组差异有显著性，P＜0.01。
结论：AgNO_3溶液有去肠粘膜作用，5％AgNO_3溶液10min处理肠段可以完全去除肠粘膜，无显著纤维化，无显著肠管挛缩，对肠膀胱成形术具有重要的临床意义。
Objective: To study the de-epithelialization effect of different concentrations of AgNO3 solutions at different time points on intestines of rabbits and explore its clinical significance on enterocystoplasy.
Methods: Forty rabbits were randomly divided into four groups (10 rabbits in each group). For each rabbit, one 15 cm segment of jejunum was isolated with intact mesentety. Intestinal anastomosis was performed to keep its continuity. The segments in group B were treated with 2g/100ml silver nitrate (AgNO3) solution for 30 minutes, while the segments in group C were treated with 5g/100ml AgNO3 solution for 10 minutes,and the mucosa of the segments in group D were stripped by surgical blade. The The segments in Group A were only treated with nomal Saline. Volume of free intestine was measured when the inner pressure was 14cmH2O(The pressure is the average pressure in normal rabbits bladder at the moment of miction. And the volume of free intestine at this pressure could reflect indirectly the compliance of the intestinal wall of the substitute bladder.). Thirty-three rabbits survived and were killed 8 weeks after operation, Volume of free intestines were measured too, and those free intestines were produced to
sections stained with HE, PAS and masson's trichrome for histopathological analysis by light microscope.
Results: At the end of the 8th week, it was displayed by HE stain that the intestinal mucosa layer in the samples of group A was integrated. In the samples of group B, there was defect in some parts of the intestinal mucosa layer and muscularis mucosa layer, and there weren't any muscularis mucosa below some parts of regeneration mucosa. In the samples of group C, there was a totally depletion of mucosa and there was a layer of false membrane constituted of cellulose, inflammatory cells and necrosis tissues over the intestinal wall. In the samples of group D, there was no mucosa layer and the submucosa layer was substituted by scar tissues. It was showed in PAS stain that there were all positive stains in group A, part of positive stains in group B and completely negative stains in group C and D. It was showed in masson's trichrome stain that there was no significant thickening of the collagen fiber in the submucosa layer in group A, B and C, and apparently thickening in group D. Comparing with group A, the
collagen fiber despot index in group B and group C had no significant change after the operation (P>0.05). But there was significant change in group D (P<0.01). And in groups A, B and C, there were no significant difference between the volumes of free
intestines during the operation and those at the 8th week after the operation (P>0.05). But in group D there were significant difference between the intestinal volumes during the operation and those at the 8th week after the operation (P<0.01).
Conclusion: AgNO3 solution can effectively inactivate intestinal mucous. Ten minutes' treatment of 5% AgNO3 solution can successfully de-epithclialize jijunal segments without creating severe retraction or fibrosis. This novel treatment can be very useful for enterocystoplasy.
引文
1. Hendren WH, Hendren EB. Bladder augmentation:experience with 129 children and young adults. J Urol, 1990,144:445-453.
2. Kajbafzadeh AM,Quinn FM,Duffy PG,et al. Augmentation cystoplasty in boys with posterior urethral valves. J Urol,1995,154:874-877.
3. Keating MA,Ludlow JK,Rich MA. Enterocystoplasty: thestar modifycation. J Urol, 1996,155:1723-1725.
4. Mcdougal WS. Metablic complications of urinary intestinal diversion. J Urol, 1992,147:1199-1208.
5. Filmer RB,Spencer JR. Malignancies in bladder augmentations and intestinal conduits. J Urol, 1990,143:671-678.
6. Kronlner KM, Casale AJ, Cain MP, et al. Bladder calculi in the pediatric augmented bladder. J Urol, 1998,160:1096-1098.
7. Motley RC,Montgomery BT, Zollman PE,et al. Augmentation cystoplasty utilizing de-epithelialized sigmoid colon:a preliminary study.J Urol, 1990,143 (6): 1257-1260.
8. Pippisalle JL,Fraga JC,Lucib A, et al. Seromuscular enterocystoplasty in dogs. JUrol, 1990,144:454-456.
9. Frey P, Lutz N ,Neubo AL .Augmentation cystoplasty using pedicled and de-epithelialized gastric patches in the minipig model. J Urol,1996,156(2): 608-613.
10. Lutz N , Frey P. Enterocystoplasty using modified pedicled, detubularized , de-epithelialized sigmoid patches in the mini-pig model. J Urol,1995,154(2):893-898.
11. Debadiola F, Manivel JC, Gonzalez R. Seromuscular enterocystoplasty in rats. J Urol,1991,146:559-562.
12. Niku SD, Scherz HC, Stein PC, Parsons CL . Intestinal de-epithelialization and augmentation cystoplasty ; an animal model. Urology, 1995,46(1): 36-39.
13. Laselhuhn GD, Kropp KA, Keck RW, Selman SH.Photochemical ablation of intestinal mucosa for bladder augmentation. J Urol ,1994,152(6) : 2267-2271.
14. Mcdougal WS. Metablic complications of urinary intest intestinal diversion. J Urol,1992, 47:1199-1208.
15. Lansdown AB, sampson B, Laupattarakasem P, Vuttivirojana A. Silver aids healing in the sterile skin wound:experimental studies in laboratory rat.Br J Dermatol. 1997;137(5):728-735.
16. Wright JB, Lam K, Burrel RE. Wound management in an era of increasing bacter ial antibiotic resistance:a rolev for topical silvertreatment .Am J Infect Control,1998;26(6):572-527.
17. Wright JB, Lam K, Hansen D, Burrel RE. Efficacy of topical silver against fungal burn wound pathogens. Am J Infect Control, 1999,27(4):344-350.
18. Hidalgo E, Dominguez C. Study of cytotoxicity mechanisms of silver nitrate in human dermal fibroblasts. Toxicol Lett, 1998,15(3): 169-179.
19. Hollinger MA, Toxicological aspect of topical silver pharmaceuticals. Crit Rev Tpxicol, 1996, 26 (3) : 255-260.
20. Liu J, Kershaw WC, Klassen CD, The protective effect of metallothionein on the toxicity various metals in rat primary hepatocyte culture .Toxicol Appl Pharmacol, 1991,107(1): 27-34.
21. Hussain S, Anner RM, Anner BM. Cysteine protects Na, K-ATPase and.isolated human lymphocytes from silver toxicity. Biochem Biophys Res Commun, 1992,189(3): 1444-1449.
22. Jansson G, Harms-Ringdahl M. Stimulating effects of mercuric and siler ions on the superoxide anion production in human polymorphonuclear leucocytes .Free Radic Res Commum, 1993,18(2),87-98.
2. Kajbafzadeh AM,Quinn FM,Duffy PG,et al. Augmentation cystoplasty in boys with posterior urethral valves. J Urol,1995,154:874-877.
3. Keating MA,Ludlow JK,Rich MA. Enterocystoplasty: thestar modifycation. J Urol, 1996,155:1723-1725.
4. Mcdougal WS. Metablic complications of urinary intestinal diversion. J Urol, 1992,147:1199-1208.
5. Filmer RB,Spencer JR. Malignancies in bladder augmentations and intestinal conduits. J Urol, 1990,143:671-678.
6. Kronlner KM, Casale AJ, Cain MP, et al. Bladder calculi in the pediatric augmented bladder. J Urol, 1998,160:1096-1098.
7. Motley RC,Montgomery BT, Zollman PE,et al. Augmentation cystoplasty utilizing de-epithelialized sigmoid colon:a preliminary study.J Urol, 1990,143 (6): 1257-1260.
8. Pippisalle JL,Fraga JC,Lucib A, et al. Seromuscular enterocystoplasty in dogs. JUrol, 1990,144:454-456.
9. Frey P, Lutz N ,Neubo AL .Augmentation cystoplasty using pedicled and de-epithelialized gastric patches in the minipig model. J Urol,1996,156(2): 608-613.
10. Lutz N , Frey P. Enterocystoplasty using modified pedicled, detubularized , de-epithelialized sigmoid patches in the mini-pig model. J Urol,1995,154(2):893-898.
11. Debadiola F, Manivel JC, Gonzalez R. Seromuscular enterocystoplasty in rats. J Urol,1991,146:559-562.
12. Niku SD, Scherz HC, Stein PC, Parsons CL . Intestinal de-epithelialization and augmentation cystoplasty ; an animal model. Urology, 1995,46(1): 36-39.
13. Laselhuhn GD, Kropp KA, Keck RW, Selman SH.Photochemical ablation of intestinal mucosa for bladder augmentation. J Urol ,1994,152(6) : 2267-2271.
14. Mcdougal WS. Metablic complications of urinary intest intestinal diversion. J Urol,1992, 47:1199-1208.
15. Lansdown AB, sampson B, Laupattarakasem P, Vuttivirojana A. Silver aids healing in the sterile skin wound:experimental studies in laboratory rat.Br J Dermatol. 1997;137(5):728-735.
16. Wright JB, Lam K, Burrel RE. Wound management in an era of increasing bacter ial antibiotic resistance:a rolev for topical silvertreatment .Am J Infect Control,1998;26(6):572-527.
17. Wright JB, Lam K, Hansen D, Burrel RE. Efficacy of topical silver against fungal burn wound pathogens. Am J Infect Control, 1999,27(4):344-350.
18. Hidalgo E, Dominguez C. Study of cytotoxicity mechanisms of silver nitrate in human dermal fibroblasts. Toxicol Lett, 1998,15(3): 169-179.
19. Hollinger MA, Toxicological aspect of topical silver pharmaceuticals. Crit Rev Tpxicol, 1996, 26 (3) : 255-260.
20. Liu J, Kershaw WC, Klassen CD, The protective effect of metallothionein on the toxicity various metals in rat primary hepatocyte culture .Toxicol Appl Pharmacol, 1991,107(1): 27-34.
21. Hussain S, Anner RM, Anner BM. Cysteine protects Na, K-ATPase and.isolated human lymphocytes from silver toxicity. Biochem Biophys Res Commun, 1992,189(3): 1444-1449.
22. Jansson G, Harms-Ringdahl M. Stimulating effects of mercuric and siler ions on the superoxide anion production in human polymorphonuclear leucocytes .Free Radic Res Commum, 1993,18(2),87-98.
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