腹泻型IBS脾虚肝郁证候与5-HT,CRH的相关性

Correlation Between D-IBS with Spleen Deficiency and Liver Stagnation Symptoms and 5-HT,CRH

目的:探讨腹泻型肠易激综合征(diarrhea-predominant irritable bowel syndrome,D-IBS)脾虚肝郁证与脑肠肽之间的关系。方法:采用灌服番泻叶联合束缚应激的方法建立腹泻型肠易激综合征大鼠模型,分别于建模前、建模第7,14,21,28天检测肠动力、内脏敏感性和5-羟色胺(5-hydroxytryptamine,5-HT),促肾上腺皮质激素释放激素(corticotropin-releasing hormone,CRH)的表达。应用Maclab/4e四导生理记录仪描记大鼠结肠运动曲线,计算结肠运动指数;采用直肠扩张腹壁撤退反射(AWR)评分评价大鼠的内脏敏感性;采用酶联免疫吸附测定法(ELISA),免疫组化法检测5-HT,CRH的含量;应用逆转录聚合酶链式反应(reverse transcription-PCR,RT-PCR)检测5-HT,CRH mRNA的表达量。结果:与建模前比较,建模第14,21,28天组大鼠结肠运动指数,AWR评分为3时的注水量、结肠和海马中5-HT,CRH及5-HT,CRH mRNA的表达均有显著差异(P<0.05,P<0.01);与建模第7天组比较,建模第21,28天组大鼠结肠运动指数、结肠运动指数变化率,AWR评分为3时的注水量、结肠和海马中5-HT,CRH及5-HT,CRH mRNA的表达差异显著(P<0.05,P<0.01);与建模第14天组比较,建模第21,28天组大鼠AWR评分为3时的注水量、结肠和海马中5-HT,CRH及5-HT,CRH mRNA的表达差异显著(P<0.05,P<0.01),建模第28天组大鼠结肠运动指数、结肠运动指数变化率有显著差异(P<0.05,P<0.01)。结论:5-HT,CRH与D-IBS脾虚肝郁证候密切相关,可能为D-IBS脾虚肝郁证病机的物质基础。

Objective： To evaluate the correlation between spleen deficiency and liver stagnation,and brain gut peptides in rats with diarrhea-predominant irritable bowel syndrome（ D-IBS）. Method： A rat model of D-IBS was established through intragastric administration with Sennae Folium decoction combined with restraint stress. Rats were divided into several groups. The modeling groups were given Sennae Folium decoction combined with restraint stress. The colonic motility,visceral sensitivity and the expression of brain gut peptides were measured at 0 d,7thd,14 thd,21std,28 thd after modeling. Maclab/4e 4-channel physiology recorder was used to record colon motion curves,and evaluate colonic motility index. The visceral sensitivity of rats was evaluated by colorectal distention,and water injection rate was recorded when the abdominal withdrawal reflex（ AWR） was scored at 3. Enzyme linked immunosorbent assay（ ELISA）,immunohistochemistry assay and reverse transcriptionpolymerase chain reaction（ RT-PCR） were used to detect the expression of brain gut peptides. Result： In 14 th d,21 std,28thd groups,the colonic motility index,the water injection rate when AWR was scored at 3,the expressions of 5-HT,CRH,5-HT and CRH mRNA in colon and hippocampus were significantly different（ P〈0. 05,P〈0. 01） from before modelling. In 21 std,28thd group,the colonic motility index,the motility index change rate,the water injection rate when AWR was scored at 3,the expressions of 5-HT,CRH,5-HT and CRH mRNA in colon and hippocampus were significantly different（ P〈0. 05,P〈0. 01） from those in the 7thd group.In 21 std,28thd groups,the water injection rate when abdominal withdrawal reflex（ AWR） was scored at 3,the expressions of 5-HT,CRH,5-HT and CRH mRNA in colon and hippocampus were significantly different（ P〈0. 05,P〈0. 01） from those in the 14 thd group. The colonic motility index and the motility index change rate in28 thd group were significantly different from those the 14 thd group（ P〈0. 05,P〈0. 01）. Conclusion： The 5-HT and CRH are closely correlated with CRH and D-IBS with spleen deficiency and liver stagnation symptoms,which might be the material basis of spleen deficiency and liver stagnation syndrome in D-IBS.