中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2010年
12期
867-871
,共5页
肠易激综合征%肥大细胞%类胰蛋白酶%内脏敏感性%P物质%降钙素基因相关肽
腸易激綜閤徵%肥大細胞%類胰蛋白酶%內髒敏感性%P物質%降鈣素基因相關肽
장역격종합정%비대세포%류이단백매%내장민감성%P물질%강개소기인상관태
Irritable bowel syndrome%Mast cells%Tryptases%Visceral sensitivity%Substance P%Calcitonin gene-related peptide
目的 观察腹泻型肠易激综合征(1BS-D)患者结肠黏膜类胰蛋白酶变化及其结肠黏膜孵育上清液对小鼠内脏感觉的影响.方法 IBS-D患者研究:肠镜检查获取24例非感染后IBS-D患者、12名健康志愿者的升结肠黏膜组织4块,采用甲苯胺蓝染色法计数黏膜肥大细胞数量、酶联免疫分析法检测结肠黏膜孵育上清液中类胰蛋白酶浓度.动物实验:56只6～8周龄C57BL/6系雄性小鼠分为7组,分别予健康小鼠升结肠黏膜培养上清液100μl(A组)、IBS-D升结肠黏膜培养上清液100 μl(B组)、IBS-D升结肠黏膜培养上清液100μl+丝氨酸蛋白酶抑制剂Fut-175(50μg/ml,C组)、0.6 ng类胰蛋白酶(D组)、2.2 ng类胰蛋白酶(E组)、2.2 ng类胰蛋白酶±Fut-175(50μg/ml,F组)和0.9%氯化钠溶液(G组)灌肠.6 h后测定结肠气囊扩张时腹壁肌电变化.免疫组化法观察腰骶部脊髓背角P物质、降钙素基因相关肽(CGRP)表达变化.结果 IBS-D患者研究:IBS-D患者升结肠肥大细胞(MC)数量为(6.33±1.64)个/高倍视野,健康对照组为(3.08±0.77)个/高倍视野,差异有统计学意义(P=0.000).IBS-D患者升结肠黏膜组织类胰蛋白酶浓度为(0.072±0.023)ng/mg组织,健康对照组为(0.026±0.014)ng/mg组织,差异有统计学意义(P=0.000).动物实验发现在气囊内压为30、45、60 mmHg时,B组及E组小鼠的腹壁肌电幅值[(27.50±5.23)、(41. 69±5.80)、(54.03±6.13)μV和(27.05±1.66)、(40.66±3.39)、(54.38±4.68)μV]较A组、C组、D组、F组、G组高(P值均＜0.05).B组及E组脊髓背角P物质和CGRP之积分吸光度值(121.72±4.67、123.54±4.32和124.69±5.06、126.32±5.09)较A组、C组、D组、F组、G组高(P值均＜0.05).结论 IBS-D患者升结肠黏膜肥大细胞数量和类胰蛋白酶含量均明显增加,而且其结肠黏膜孵育上清液可导致小鼠内脏感觉过敏,提示类胰蛋白酶可能参与IBS-D患者内脏敏感性的改变.
目的 觀察腹瀉型腸易激綜閤徵(1BS-D)患者結腸黏膜類胰蛋白酶變化及其結腸黏膜孵育上清液對小鼠內髒感覺的影響.方法 IBS-D患者研究:腸鏡檢查穫取24例非感染後IBS-D患者、12名健康誌願者的升結腸黏膜組織4塊,採用甲苯胺藍染色法計數黏膜肥大細胞數量、酶聯免疫分析法檢測結腸黏膜孵育上清液中類胰蛋白酶濃度.動物實驗:56隻6～8週齡C57BL/6繫雄性小鼠分為7組,分彆予健康小鼠升結腸黏膜培養上清液100μl(A組)、IBS-D升結腸黏膜培養上清液100 μl(B組)、IBS-D升結腸黏膜培養上清液100μl+絲氨痠蛋白酶抑製劑Fut-175(50μg/ml,C組)、0.6 ng類胰蛋白酶(D組)、2.2 ng類胰蛋白酶(E組)、2.2 ng類胰蛋白酶±Fut-175(50μg/ml,F組)和0.9%氯化鈉溶液(G組)灌腸.6 h後測定結腸氣囊擴張時腹壁肌電變化.免疫組化法觀察腰骶部脊髓揹角P物質、降鈣素基因相關肽(CGRP)錶達變化.結果 IBS-D患者研究:IBS-D患者升結腸肥大細胞(MC)數量為(6.33±1.64)箇/高倍視野,健康對照組為(3.08±0.77)箇/高倍視野,差異有統計學意義(P=0.000).IBS-D患者升結腸黏膜組織類胰蛋白酶濃度為(0.072±0.023)ng/mg組織,健康對照組為(0.026±0.014)ng/mg組織,差異有統計學意義(P=0.000).動物實驗髮現在氣囊內壓為30、45、60 mmHg時,B組及E組小鼠的腹壁肌電幅值[(27.50±5.23)、(41. 69±5.80)、(54.03±6.13)μV和(27.05±1.66)、(40.66±3.39)、(54.38±4.68)μV]較A組、C組、D組、F組、G組高(P值均＜0.05).B組及E組脊髓揹角P物質和CGRP之積分吸光度值(121.72±4.67、123.54±4.32和124.69±5.06、126.32±5.09)較A組、C組、D組、F組、G組高(P值均＜0.05).結論 IBS-D患者升結腸黏膜肥大細胞數量和類胰蛋白酶含量均明顯增加,而且其結腸黏膜孵育上清液可導緻小鼠內髒感覺過敏,提示類胰蛋白酶可能參與IBS-D患者內髒敏感性的改變.
목적 관찰복사형장역격종합정(1BS-D)환자결장점막류이단백매변화급기결장점막부육상청액대소서내장감각적영향.방법 IBS-D환자연구:장경검사획취24례비감염후IBS-D환자、12명건강지원자적승결장점막조직4괴,채용갑분알람염색법계수점막비대세포수량、매련면역분석법검측결장점막부육상청액중류이단백매농도.동물실험:56지6～8주령C57BL/6계웅성소서분위7조,분별여건강소서승결장점막배양상청액100μl(A조)、IBS-D승결장점막배양상청액100 μl(B조)、IBS-D승결장점막배양상청액100μl+사안산단백매억제제Fut-175(50μg/ml,C조)、0.6 ng류이단백매(D조)、2.2 ng류이단백매(E조)、2.2 ng류이단백매±Fut-175(50μg/ml,F조)화0.9%록화납용액(G조)관장.6 h후측정결장기낭확장시복벽기전변화.면역조화법관찰요저부척수배각P물질、강개소기인상관태(CGRP)표체변화.결과 IBS-D환자연구:IBS-D환자승결장비대세포(MC)수량위(6.33±1.64)개/고배시야,건강대조조위(3.08±0.77)개/고배시야,차이유통계학의의(P=0.000).IBS-D환자승결장점막조직류이단백매농도위(0.072±0.023)ng/mg조직,건강대조조위(0.026±0.014)ng/mg조직,차이유통계학의의(P=0.000).동물실험발현재기낭내압위30、45、60 mmHg시,B조급E조소서적복벽기전폭치[(27.50±5.23)、(41. 69±5.80)、(54.03±6.13)μV화(27.05±1.66)、(40.66±3.39)、(54.38±4.68)μV]교A조、C조、D조、F조、G조고(P치균＜0.05).B조급E조척수배각P물질화CGRP지적분흡광도치(121.72±4.67、123.54±4.32화124.69±5.06、126.32±5.09)교A조、C조、D조、F조、G조고(P치균＜0.05).결론 IBS-D환자승결장점막비대세포수량화류이단백매함량균명현증가,이차기결장점막부육상청액가도치소서내장감각과민,제시류이단백매가능삼여IBS-D환자내장민감성적개변.
Objective To observe the variation of tryptase expression in the colonic mucosa of patients with diarrhea-predominant irritable bowel syndrome(IBS-D) and the effects of their colonic mucosa cultured supernatants on mice visceral sensitivity. Methods The IBS-D patients study: of twenty-four patients with IBS-D and 12 healthy volunteers, four mucosal biopsies of each individual were collected at ascending colon under colonoscopy. The quantity of mucosal mast cells were detected by toluidine blue stains, and the tryptase concentration of colonic mucosa cultured supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Animal experiments: fifty-six male six to eight weeks old C57BL/6 mice were divided into 7 groups and then were administrated with healthy volunteer's ascending colonic mucosa-cultured supernatants 100 μl (group A), IBS-D patient' s ascending colonic mucosa-cultured supernatants 100 μl (group B), IBS-D ascending colonic mucosacultured supernatants 100 μl with Fut-175(50 μg/ml, group C), tryptase with different concentration:0.6 ng (group D), 2.2 ng (group E), tryptase 2.2 ng with Fut-175 (group F), and 0.9% sodium chloride as control (group G) respectively. After six hours, the abdominal myoelectric activities (AMAs) were recorded during colorectal balloon distension (CRD), and the expressions of substance P (SP), Calcitonin gene-related peptide (CGRP) in spinal dorsal horn of lumbosaeral region were detected by immunohistochemistry. Results The number of mast cells in the ascending colon of patients with IBS-D [6.33 ± 1.64) / HP] was higher than that in controls [(3.08 ± 0.77 ) / HP], the difference was statistical significant (P = 0.000). The tryptase concentration of ascending colonic mucosa cultured supernatants in IBS-D patients [(0. 072 ± 0. 023) ng/mg] was higher than that in controls [(0.026 ± 0.014) ng/mg], the difference was statistical significant ( P = 0. 000). In animal experiments, when the inside pressure of balloon was 30, 45, 60 mmHg, respectively, the AMAs value in group B [(27.50±5.23), (41.69±5.80), (54. 03±6.13) μV, respectively] and group E [(27. 05±1.66), (40. 66±3.39), (54. 38± 4.68) μV, respectively] were significantly higher than those in group A, group C, group D, group F and group G (P＜0. 05, respectively). The average integrated option densities (IOD) of SP and CGRP in spinal dorsal horn in groupB (121.72 ± 4.67,123.54±4.32)and E (124.69±5.06,126.32±5.09) were higher than in group A, group C, group D,group F and group G (P＜0.05, respectively). Conclusion The quantity of mast cell and the content of tryptase in the ascending colonic mucosa of IBS-D patients increased significantly. And their colonic mucosa cultured supernatants caused the mice visceral hypersensitivity, which indicated that tryptase may involved in the variation of visceral sensitivity in IBS-D patients.
