| ObjectiveDepression is closely associated with Traditional Chinese Medicinelily disease, the intestinal abnormal reaction is one of lily disease symptoms, clinical depression is often associated with irritable bowel syndrome (IBS) gastrointestinal complications, brain-gut interaction theory reveals the causes of depression more associated with IBS, but its scientific theory has not yet been elucidated. This study based on the brain-gut interaction theory preliminarily study lily saponin/timosaponin/lily timosaponin effect on the treatment of depression accompanied IBS and its related mechanism.Methods1 Lily-Timosaponin treatment of depression accompanied IBS model rat external index detectionAdult male SD rats, according to the weight and the open-field experiment choose similar rats, adopts the method of half randomly divided into Normal control group, Model group, Lily saponin group, Timosaponingroup, Lily-Timosaponin group, depression positive medicine FluoxetineHydrochloridegroup, IBSpositive medicinePinaverium Bromide Dicetelgroup. Normal don’t do any processing, Model and the rest of the five groups of rats, to solitary, clamping tail stress, wrap-restraint stress, chronic unpredictable mild stress induction model, All the drugs were administered gastrogavage once daily, for six weeks. Testing different experimental rat body weight, the open-field experiment score, sucrose water consumption, AWR score, and the quantity of defecation.2 Lily-Timosaponin in depression accompanied IBS model rat Efficacy of indicators and its mechanism preliminarily studyUsing ELISA method to detect rats serum ACTH, DA, CGRP and VIP content;to detect plasmaCRF, SP, IL-1 beta, IL-6 content; to detect rats in brain tissue and colon tissue 5-HTcontent. Application HE conventional dyeing and aldehyde fuchsin-orange G special testing rats form and quantity of thecolon tissuemast cells change;Using the fluorescent quantitative PCR detection rats in the hypothalamus tissue CRF mRNA and its receptorCRFRlmRNA, CRFR2 mRNA relativeexpression changes.3 test and verify depression accompanied IBS model rats differentially expressed genesUsingagilent gene expression chipto detect rat’s blood differences gene expression, comparison of between Normal, Model, LS and LT gene difference expression. Screening drug intervention significant pullback two genes (Rgsl9 an Ormdl3), and review the relatedliteraturefind outdifferences genes functional that the use of real-time PCR technology to verify the blood, the hypothalamus tissue and colon tissue of Rgs19 mRNA and Ormd13 mRNA differentially expressionofreliability. Results1 modeling six weeks after the treatment, compared with the Normal, the weight of Model rats, Locomotor activity, sucrose waterconsumptionand visceral sensitivity have significant differences (P＜0.01), characterized byweightlow growth, behavior movementreduction,less the amount of sucrose water consumption, defecate number increase, visceral sensitivity increased significantly. Compared with the Model, external indicators the saponin group have significantly improved, among them Lily-timosaponin superior to single flavor saponins effect.2 compared with Normal, Model ratsthe concentration ofserum ACTH, CGRP and VIPandplasma CRF, SP, IL-1 beta, IL-6 levels increased significantly (P＜0.01); the saponin group can significantly reducerat’sthe content of serum ACTH, CGRP and VIP and plasma CRF, SP, IL-1 beta, IL-6 (P＜0.05 or P＜ 0.01);Compared with Normal, Model DA content in serum of rats decreased significantly (P＜0.01), the saponin group were significantly higher DA content in serum of rats (P＜0.05);Compared with the Normal, in colon tissue of Model rats had the content of 5-HT has obvious changes, and colon tissue of mast cells number and degranulation significantly increased;The saponin significantly reversed theabnormal changes of5-HT and mast cells.By real-time PCR detection result showed that compared with Normal, Model rat’s hypothalamus tissue tissue CRF mRNA and its receptor CRFR1 mRNA expression was significantly increased (P＜0.01), LT can significantly reduce the expression (P＜0.01); compared with Normal, Model rat’s hypothalamus tissue and colon tissue CRFR2 mRNA expression was significantly lower (P＜0.05), no statistically significant differences amongthe saponin groupwith Model.3 This study for the first time in depression accompanied IBS rats model use Agilent gene expression chip. Screening drugs callback significantly differentially expressed genes, found lily saponin and lily timosaponin common callback difference genes fold-change more than 1.50 times genes has eight:Ormd13/Rgsl9/Ptprn2/Cym/Tmem47/Rpl37/Zcrb/Ext13. This study verified the Model compared with Normaldifferencehigher multiples of two genes Rgs19 mRNA,Ormdl3 mRNA expression results consistent with the results of Agilent gene expression chip. ConclusionUsing chronic stressors can be successfully induced depression accompanied IBS rat model. The open-field test, sucrose water consumption test, AWR score test and defecation grains changes can evaluate the effectiveness of the model. Lily saponin, timosaponin and Lily timosaponin treatment of depression accompanied IBS may be related to the reversal of the brain-gut interaction mechanisms HAP axis abnormal ACTH, CRF levels, reversing monoamine neurotransmitters DA,5-HT content reverse nerve immune factors IL-1β, IL-6 levels, and can reduce the number of colon tissue mast cells; while its mechanism of action may also be associated with the reversal of the brain-gut peptide CGRP, VIP, SP,5-HT content and CRF mRNA and its receptor CRFR1 mRNA, CRFR2 mRNA expression related. Blood, hypothalamus and colon tissue Rgsl9 mRNA,Ormd13 mRNA differential expression may be involved in the pathogenesis of depression accompanied IBS. Lily saponin, timosaponin and lily timosaponin can effectively correct the abnormal state of the gene expression may be one of the mechanisms of the disorder medication. |
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