电针对大鼠肠易激综合征及抑郁情绪的改善作用及机制研究

Effect and Mechanism of EA in Improving IBS Accompanied by Depression

目的:观察中医针刺疗法中电针“天枢”“上巨虚”方法改善肠易激综合征(IBS)伴抑郁症的作用以及对该模型大鼠结肠组织中瞬时受体电位香草酸亚型1(TRPV1)、酪氨酸激酶受体A(TrkA)、神经生长因子(NGF)、Rho相关激酶ROCK、Ras同源基因家族蛋白A(RHOA)、海马组织中NFκB和SIRT2蛋白表达的影响,探究其作用机制。方法:36只SPF级雄性SD大鼠随机分为健康空白组、模型组、电针组和西药组4组,每组9只。后3组通过二硝基苯磺酸(DNBS)灌肠和慢性束缚应激方法建立IBS伴抑郁症模型。电针组用电针刺激大鼠双侧“天枢”“上巨虚”穴位,西药组用匹维溴铵混悬液(18 mg/kg)灌胃处理,分别持续7 d,1次/d。造模前后及治疗干预后记录各组大鼠粪便含水量、糖水偏嗜程度以及高架十字迷宫测试。ELISA技术检测血清炎症因子的指标。采用免疫组化技术检测大鼠海马组织NFκB、SIRT2蛋白以及结肠组织BDNF、SIRT2、NFκB、CREB蛋白的表达和分布情况,Western Blot技术检测结肠组织中TRPV1、TrkA、NGF、ROCK及RHOA的蛋白表达变化。结果:干预后与模型组比较,电针组和西药组大鼠粪便含水量减少,差异具有统计学意义(P<0.01或P<0.05),糖水消耗增加,差异具有统计学意义(P<0.01),开放臂停留时间百分比增加,差异具有统计学意义(P<0.05),血清炎症因子降低,结肠组织TRPV1、TrkA、NGF、ROCK及RHOA蛋白表达降低,差异具有统计学意义(P<0.01或P<0.05),海马组织NFκB阳性表达降低,差异具有统计学意义(P<0.05),SIRT2阳性表达升高,差异具有统计学意义(P<0.05)。结论:电针“天枢”“上巨虚”可有效改善IBS-D模型大鼠的胃肠症状,缓解大鼠伴随的抑郁情绪和炎性因子水平,其机制可能与降低模型大鼠结肠组织TRPV1、TrkA、NGF、ROCK及RHOA蛋白表达有关。

Objective:To observe the effect of electro-needling Tianshu(ST25)and Shangjuxu(ST37)in improving irritable bowel syndrome(IBS)accompanied by depression and its influence to protein expressions of TRPV1,TrkA,NGF,ROCK and RHOA in IBS accompanied by depression,and to explore its action mechanism.Methods:36 male SD rats were randomly divided into the blank group,the model group,the electro-acupuncture(EA)group and the Western medication group,with 9 rats in each group.IBS accompanied by depression model was established by Dinitrobenzene Sulfonic Acid(DNBS)enema combined with chronic restraint stress.The EA group was treated with electro-needling bilateral ST25 and ST37.The Western medication group was treated with Pinaverium Bromide Suspension(18 mg/kg)by gavage,once per day for 7 days.Fecal water content and sucrose consumption test were recorded.For the depressive behavior assessment,the elevated plus maze test was used and the open-arm residence time ratio was recorded.ELISA assay was used for measuring inflammatory factor levels.Immunohistochemistry was utilized to detect the protein expressions and distributions of NFκB and SIRT2 in hippocampus and the proteins of BDNF,SIRT2,NFκB and CREB in rat colon tissues.Western Blot technology was used to detect the protein expressions of TRPV1,TrkA,NGF,ROCK and RHOA in colon tissues.Results:Compared with those in the model group,the water content of the feces was decreased(P<0.01,P<0.05),the consumption of sugar water was increased(P<0.01),the open-arm residence time ratio was increased(P<0.05),the serum levels of inflammatory factors and the protein expressions of TRPV1,TrkA,NGF,ROCK and RHOA in colon tissues were reduced(P<0.01,P<0.05),the positive expression of NFκB in hippocampus was decreased(P<0.05),and the positive expression of SIRT2 was increased in the EA group and the Western medication group(P<0.05).Conclusion:Electro-needling ST25 and ST37 can effectively improve the gastrointestinal symptoms of IBS accompanied by depression,and relieve the accompanied depression and reduce inflammatory factor levels,which may be related to the regulation on reducing the protein expressions of TRPV1,TrkA,NGF,ROCK and RHOA in colon tissues in rats.