肠易激综合征结肠粘膜AQP8表达与VIP、SS的表达研究
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摘要
目的:本课题主要通过测定肠易激综合征(IBS)升、降结肠粘膜水通道蛋白8(AQP8)和胃肠激素血管活性肠肽(VIP)、生长抑素(SS)的表达变化。探讨肠易激综合征水通道蛋白8和VIP、SS在IBS发病机制中所起的作用,揭示肠易激综合征的病因和可能的发病机制。
方法:根据肠易激综合征罗马Ⅲ诊断标准收集山西省人民医院2006年11月至2007年2月腹泻型(IBS-D)患者31例、便秘型(IBS-C)患者25例,以及健康志愿者15例。通过免疫组织化学方法和逆转录聚合酶链反应(RT-PCR)方法定性和定量分析研究结肠粘膜AQP8表达量以及血管活性肠肽(VIP)、生长抑素(SS)在升、降结肠粘膜的表达变化。
结果:
1、AQP8在人结肠粘膜广泛表达,无论正常对照组及IBS组。AQP8主要表达在结肠上皮细胞的顶膜和基侧膜,部分结肠上皮细胞胞浆也有表达。
2、AQP8在正常对照组、IBS—D和IBS—C组升、降结膜粘膜均有表达,与正常对照组相比,IBS—D组AQP8表达明显降低(p＜0.05),IBS—C组AQP8表达显著升高(p＜0.05),并且IBS—C组AQP8表达明显高于IBS—D组(p＜0.05)。但在三组间升、降结肠粘膜AQP8比较中,仅IBS—D组升结肠表达明显高于降结肠(p＜0.05),而正常对照组和IBS—C组差异不显著(p＞0.05)。
3、胃肠激素VIP、SS在人升、降结肠粘膜均有表达,主要表达于结肠粘膜的粘膜层和固有层的内分泌细胞胞浆内。
4、VIP在正常对照组、IBS—D和IBS—C组升、降结膜粘膜都有表达,与正常对照组相比,IBS—D组和IBS—C组VIP表达水平均明显升高(p＜0.05),而且IBS—C组VIP表达水平明显高于IBS—D组(p＜0.05),但各组间升、降结肠粘膜VIP表达无显著差异(p＞0.05)。
5、SS在正常对照组、IBS—D和IBS—C组升、降结膜粘膜都有表达,与正常对照组相比,IBS—D组和IBS—C组SS表达水平均升高,IBS—C组SS表达水平显著升高(p＜0.05),而IBS—D组SS表达水平升高不显著(p＞0.05)。同时IBS—C组SS表达水平明显高于IBS—D组(p＜0.05)。而各组间升、降结肠粘膜SS表达无显著差异(p＞0.05)。
结论:
1、AQP8、VIP和SS在人升、降结肠粘膜都有表达。
2、AQP8在IBS患者结肠粘膜上皮细胞水的转运过程中起重要作用,参与结肠粘膜水的调节,可能是IBS产生腹泻和便秘的重要原因之一。。
3、胃肠激素VIP、SS在IBS病因和发病机制中起重要作用,可能成为神经—免疫—内分泌网络调节中的重要一环。
Objective: To investigate the expression of aquaporin-8 (AQP8) and Vasoactive Intestinal Peptide(VIP), somatostatin (SS) protein in human colonic mucosa with irritable bowel syndrome(IBS), and elucidate their possible roles in the pathogenesis of IBS
Methods: Based on RomeⅢcriteria, 56 IBS patients, including 31 IBS-D, 25 IBS-C and 15 healthy subjects were enrolled from November 2006 to February 2007 in Shanxi Provincial People's Hospital in the study. The expression of AQP8mRNA in colonic mucosa was measured by transcription polymerase chain reaction(RT-PCR), and AQP8、VIP、SS in ascend and descend colonic mucosa were measured by immunohistochemistry(IHC).
Results:
1. There were all expression of AQP8 in human colonic mucosa with IBS and normal group. AQP8 was strongly expressed in the apical membrane and the basolateral membrane, and there was litter expression in cytoplasm in colonic epithelial cells.
2. There were all expression of AQP8 in ascend and descend colonic mucosa with IBS-D, IBS-C and normal group. Compared with normal control group, the level of AQP8 in IBS-D was significantly reduced and the level of AQP8 in IBS-C was obviously increased(p<0.05), and the expression of AQP8 in IBS-C group was significantly higher than that of the IBS-D group(p<0.05). However, there was no significant difference in ascend and descend colonic mucosa of normal group and IBS-C group(p>0.05). only in IBS-D group, the expression of AQP8 of the ascend colon was significantly higher than that of the descending colon(p<0.05).
3. There were all expression of the gut hormone of VIP, SS in human ascend and descend colonic mucosa with IBS and normal group. VIP and SS was mainly expressed in the endocrine cells of the mucous layer and lamina propria of the colonic mucosa.
4. There were all expression of VIP in ascend and descend colonic mucosa with IBS-D, IBS-C and normal group. Compared with normal control group, the level of VIP in IBS-D and IBS-C were all significantly increased(p<0.05), and the expression of VIP in IBS-C group was significantly higher than that of the IBS-D group(p<0.05). However, there was no significant difference in ascend and descend colonic mucosa of each group(p>0.05).
5. There were all expression of SS in ascend and descend colonic mucosa with IBS-D, IBS-C and normal group. Compared with normal control group, the leyel of SS in IBS-C was significantly increased(p<0.05), the expression of SS in IBS-C group was significantly higher than that of the IBS-D group and normal group(p<0.05), and the level of SS in IBS-D was only slightly increased but not significantly in IBS-D group(p>0.05). However, there was also no significant difference in ascend and descend colonic mucosa of each group(p>0.05).
Conclusion:
1、There were all expression of AQP8, VIP and SS in human ascend and descend colonic mucosa with IBS and normal group.
2、AQP8 may play a major role in water movement through the colonic epithelial cells, and probably involve in the regulation of water in colonic mucosa with IBS. The change of the AQP8 may be one of the factors which result in diarrhea or constipation in IBS.
3、The gut hormones including VIP and SS may play an important role in the pathogenesis of IBS. It is probably becoming new proof for the neuro-endocrine-immune network regulation theory.
方法:根据肠易激综合征罗马Ⅲ诊断标准收集山西省人民医院2006年11月至2007年2月腹泻型(IBS-D)患者31例、便秘型(IBS-C)患者25例,以及健康志愿者15例。通过免疫组织化学方法和逆转录聚合酶链反应(RT-PCR)方法定性和定量分析研究结肠粘膜AQP8表达量以及血管活性肠肽(VIP)、生长抑素(SS)在升、降结肠粘膜的表达变化。
结果:
1、AQP8在人结肠粘膜广泛表达,无论正常对照组及IBS组。AQP8主要表达在结肠上皮细胞的顶膜和基侧膜,部分结肠上皮细胞胞浆也有表达。
2、AQP8在正常对照组、IBS—D和IBS—C组升、降结膜粘膜均有表达,与正常对照组相比,IBS—D组AQP8表达明显降低(p＜0.05),IBS—C组AQP8表达显著升高(p＜0.05),并且IBS—C组AQP8表达明显高于IBS—D组(p＜0.05)。但在三组间升、降结肠粘膜AQP8比较中,仅IBS—D组升结肠表达明显高于降结肠(p＜0.05),而正常对照组和IBS—C组差异不显著(p＞0.05)。
3、胃肠激素VIP、SS在人升、降结肠粘膜均有表达,主要表达于结肠粘膜的粘膜层和固有层的内分泌细胞胞浆内。
4、VIP在正常对照组、IBS—D和IBS—C组升、降结膜粘膜都有表达,与正常对照组相比,IBS—D组和IBS—C组VIP表达水平均明显升高(p＜0.05),而且IBS—C组VIP表达水平明显高于IBS—D组(p＜0.05),但各组间升、降结肠粘膜VIP表达无显著差异(p＞0.05)。
5、SS在正常对照组、IBS—D和IBS—C组升、降结膜粘膜都有表达,与正常对照组相比,IBS—D组和IBS—C组SS表达水平均升高,IBS—C组SS表达水平显著升高(p＜0.05),而IBS—D组SS表达水平升高不显著(p＞0.05)。同时IBS—C组SS表达水平明显高于IBS—D组(p＜0.05)。而各组间升、降结肠粘膜SS表达无显著差异(p＞0.05)。
结论:
1、AQP8、VIP和SS在人升、降结肠粘膜都有表达。
2、AQP8在IBS患者结肠粘膜上皮细胞水的转运过程中起重要作用,参与结肠粘膜水的调节,可能是IBS产生腹泻和便秘的重要原因之一。。
3、胃肠激素VIP、SS在IBS病因和发病机制中起重要作用,可能成为神经—免疫—内分泌网络调节中的重要一环。
Objective: To investigate the expression of aquaporin-8 (AQP8) and Vasoactive Intestinal Peptide(VIP), somatostatin (SS) protein in human colonic mucosa with irritable bowel syndrome(IBS), and elucidate their possible roles in the pathogenesis of IBS
Methods: Based on RomeⅢcriteria, 56 IBS patients, including 31 IBS-D, 25 IBS-C and 15 healthy subjects were enrolled from November 2006 to February 2007 in Shanxi Provincial People's Hospital in the study. The expression of AQP8mRNA in colonic mucosa was measured by transcription polymerase chain reaction(RT-PCR), and AQP8、VIP、SS in ascend and descend colonic mucosa were measured by immunohistochemistry(IHC).
Results:
1. There were all expression of AQP8 in human colonic mucosa with IBS and normal group. AQP8 was strongly expressed in the apical membrane and the basolateral membrane, and there was litter expression in cytoplasm in colonic epithelial cells.
2. There were all expression of AQP8 in ascend and descend colonic mucosa with IBS-D, IBS-C and normal group. Compared with normal control group, the level of AQP8 in IBS-D was significantly reduced and the level of AQP8 in IBS-C was obviously increased(p<0.05), and the expression of AQP8 in IBS-C group was significantly higher than that of the IBS-D group(p<0.05). However, there was no significant difference in ascend and descend colonic mucosa of normal group and IBS-C group(p>0.05). only in IBS-D group, the expression of AQP8 of the ascend colon was significantly higher than that of the descending colon(p<0.05).
3. There were all expression of the gut hormone of VIP, SS in human ascend and descend colonic mucosa with IBS and normal group. VIP and SS was mainly expressed in the endocrine cells of the mucous layer and lamina propria of the colonic mucosa.
4. There were all expression of VIP in ascend and descend colonic mucosa with IBS-D, IBS-C and normal group. Compared with normal control group, the level of VIP in IBS-D and IBS-C were all significantly increased(p<0.05), and the expression of VIP in IBS-C group was significantly higher than that of the IBS-D group(p<0.05). However, there was no significant difference in ascend and descend colonic mucosa of each group(p>0.05).
5. There were all expression of SS in ascend and descend colonic mucosa with IBS-D, IBS-C and normal group. Compared with normal control group, the leyel of SS in IBS-C was significantly increased(p<0.05), the expression of SS in IBS-C group was significantly higher than that of the IBS-D group and normal group(p<0.05), and the level of SS in IBS-D was only slightly increased but not significantly in IBS-D group(p>0.05). However, there was also no significant difference in ascend and descend colonic mucosa of each group(p>0.05).
Conclusion:
1、There were all expression of AQP8, VIP and SS in human ascend and descend colonic mucosa with IBS and normal group.
2、AQP8 may play a major role in water movement through the colonic epithelial cells, and probably involve in the regulation of water in colonic mucosa with IBS. The change of the AQP8 may be one of the factors which result in diarrhea or constipation in IBS.
3、The gut hormones including VIP and SS may play an important role in the pathogenesis of IBS. It is probably becoming new proof for the neuro-endocrine-immune network regulation theory.
引文
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