石斛碱对肝郁脾虚型肠易激综合征小鼠GFAP、NGF、BDNF 表达及内脏敏感性的影响

Effects of dendrobine on visceral sensitivity associated GFAP,NGF,BDNF ex-pressions in mice with irriTable bowel syndrome of liver Qi-Stagnation&Spleen-Deficiency Pattern

目的探讨石斛碱对肝郁脾虚型肠易激综合征(IBS)小鼠胶质纤维酸性蛋白(GFAP)、神经生长因子(NGF)、脑源性神经营养因子(BDNF)表达及对内脏敏感性的影响。方法C57BL/6小鼠随机分为正常对照组、模型组、石斛碱各剂量组、匹维溴铵阳性组,除正常对照组外,其余各组小鼠均采用番泻叶+束缚应激法进行IBS造模;用烘干法测小鼠粪便含水率;采用直肠气囊扩张(CRD)+腹壁撤退反射(AWR)评分法行小鼠内脏敏感性测量;采用酶联免疫吸附法(ELISA)检测小鼠血清5-羟色胺(5-HT)和P物质(SP)水平,血清NGF、BDNF、GFAP表达;以蛋白免疫印迹法(Western blot)检测结肠组织中NGF、BDNF表达。结果与正常对照组比较,模型组小鼠粪便含水率、AWR评分、血清5-HT、SP水平、血清NGF、BDNF、GFAP表达、结肠组织NGF、BDNF表达均升高;与模型组比较,石斛碱各剂量组和匹维溴铵阳性组小鼠粪便含水率、AWR评分、血清5-HT、SP水平、血清NGF、BDNF、GFAP表达、结肠组织NGF、BDNF蛋白表达均降低;石斛碱各组呈剂量依赖性,石斛碱高剂量组与匹维溴铵阳性组各指标比较,差异无统计学意义。结论石斛碱可能通过减少5-HT、SP释放,抑制GFAP、NGF、BDNF表达,降低内脏敏感性,改善肝郁脾虚型IBS模型小鼠疼痛、腹泻等症状。

AIM To investigate the effects of dendrobine on the visceral sensitivity associated expressions of glial fibrillary acidic protein(GFAP),nerve growth factor(NGF)and brain-derived neurotrophic factor(BDNF)in mice with irriTable bowel syndrome(IBS)of liver Qi-Stagnation&Spleen-Deficiency Pattern.METHODS C57BL/6 mice randomly divided into normal control group,model group,low-dose,medium-dose and high-dose dendrobine groups,and pinaverium bromide positive control group were induced IBS model by folium sennae+restraint stress method except the control group.Subsequently saline was dosed upon the normal control group and the model group,while other corresponding doses of drugs were administered to the other groups.Mice were subjec-ted to the fecal water content measurement by drying method;and the determination of visceral sensitivity by color-ectal distension(CRD)+abdominal withdrawal reflex(AWR)scoring method.Serum and colon tissues were pro-cured for the detections of serum contents of 5-hydroxytryptamine(5-HT),substance P(SP),NGF,BDNF and GFAP by enzyme-linked immunosorbent assay(ELISA),and colon expressions of NGF and BDNF by Western blot.RESULTS Compared with the normal control group,the model group shared significantly higher fecal water content,AWR score,contents of serum 5-HT,SP,and increased expressions of NGF,BDNF and GFAP,protein expressions of NGF and BDNF in colon tissues;while dendrobine groups and positive pinaverium bromide group significantly reversed the aforementioned results.Dendrobine performed in a dose-dependent manner,and no significant difference was observed between the high dose dendrobine and pinaverium bromide.CONCLUSION Dendrobine may reduce the release of nociceptive neurotransmitters,inhibit the visceral sensitivity associated expressions of GFAP,NGF and BDNF,and thus improve the pain and diarrhea of mice with IBS of liver Qi-Stagnation&Spleen-Deficiency Pattern.