CN106389395A

CN106389395A - Use of resveratrol in preparation of drug for treating irritable bowel syndrome (IBS)

Info

Publication number: CN106389395A
Application number: CN201610919143.1A
Authority: CN
Inventors: 余颖聪; 徐英
Original assignee: Wenzhou Peoples Hospital
Current assignee: Wenzhou Peoples Hospital
Priority date: 2016-10-21
Filing date: 2016-10-21
Publication date: 2017-02-15

Abstract

The invention discloses a use of resveratrol in preparation of a drug for treating irritable bowel syndrome (IBS). A study finds that an IBS patient has symptoms of high intestinal sensitivity and intestinal motility reduction and resveratrol can significantly reduce high intestinal sensitivity of IBS patients and can significantly reverse the weakening of intestinal power. The invention provides a basis for use of resveratrol as a safe and effective drug for treating IBS.

Description

Application in preparation treatment irritable bowel syndrome medicine for the resveratrol

Technical field

The invention belongs to field of medicaments, it is related to a kind of new medicine use of resveratrol and in particular to resveratrol is in system Application in standby treatment irritable bowel syndrome medicine.

Background technology

Irritable bowel syndrome (irritable bowel syndrome, IBS) is common gastrointestinal function disease, with Stomachache or abdominal discomfort, companion's bowl evacuation habit and stool are characterized extremely, and prevalence is about 11%.Because disease is in slow Property and repeated relapsing, IBS patients ' life quality is generally reduced, and its high prevalence and the very long course of disease cause great society And financial burden.So its pathogenesis not yet illustrates completely.Instestinal motility is abnormal and the high sensitivity of internal organs is two pathology main greatly Physiological foundation.

Resveratrol (Resveratrol) is a kind of polyphenols being present in natural plants, is mainly derived from Portugal The plants such as grape (red wine), Rhizoma Polygoni Cuspidati, Semen arachidis hypogaeae, Fructus Mori.Resveratrol has fat-soluble, the features such as easily pass through blood brain barrier.This Invention is tested to IBS animal model by resveratrol, in conjunction with the detection such as Instestinal motility and visceral sensitivity, finds white Herba chenopodii Reed alcohol can be used in the medicine of preparation treatment irritable bowel syndrome as effective ingredient.

Content of the invention

Present invention aims to the deficiencies in the prior art, provide a kind of new medicine use of resveratrol, that is,：In vain Application in preparation treatment irritable bowel syndrome medicine for the veratryl alcohol.

By application in preparation treatment irritable bowel syndrome medicine for the resveratrol, can effectively reduce the intestinal of IBS patient Road hypersensitivity simultaneously can significantly reverse weakening of Instestinal motility, and resveratrol can be applicable to the treatment IBS of brain intestinal double target spot effect Safely and effectively medicine.

Brief description

Fig. 1 is the impact to IBS model mice body weight for the resveratrol.

Fig. 2 is the impact to IBS model mice visceral sensitivity for the resveratrol.

Fig. 3 is the impact to IBS model mice food coloring ink propulsive rate for the resveratrol.

Specific embodiment

With reference to specific pharmacological evaluation and result, the present invention will be further described.

First, materials and methods

(1) experiment material

1. laboratory animal

Male ICR mouse, body weight 26-33g.Mouse feeder is in USA New York state university Buffalo school district Animal House.Raise Foster condition is as follows：23～25 DEG C of temperature, humidity 40%～50%, natural lighting, ad lib, water inlet.All mices are prior to raising Adapt to 3 days in foster environment, then start zoopery again.Zoopery obtains relevant medical Ethics Committee on request and agrees to.

2. experimental drug

Resveratrol (resveratrol), rolipram (rolipram), desmethylimipramine (despramin) He Dixi Dissolve (diazepam) and be purchased from Sigma Co., USA.All reagent are all dissolved in the physiology aqueous solution of 5%DMSO.Wherein, depressed Related positive control drug is imipramine, and the related positive control drug of anxiety is diazepam, and rolipram is phosphodiesterase 4 (PDE4) inhibitor, the latter will be further used as mechanism target spot and carry out preliminary comparative study.

3. experiment equipment

Catheter.

(2) experimental technique

A. animal selects and packet

ICR mice is randomly divided into 8 groups, every group 12.Normal group, slowly acute joint Stress model group, resveratrol Effect group (point basic, normal, high three various dose, respectively 2.5,5,10mg/kg, gavage) and positive controls (cough up Li Pu Orchid, 1.25mg/kg, lumbar injection, desmethylimipramine, 10mg/kg, lumbar injection, diazepam, 0.5mg/kg, lumbar injection). In addition to Normal group, remaining group all carry out slowly acute joint stress, that is, setting up classical acute slowly joint stress mice mould Intend IBS model.

B. experimental procedure

1. set up slowly acute joint stress Model with Irritable Bowel Syndrome (chronic acute combining stress, CACS)

IBS rat model adopts acute slowly joint Stress model (the chronic acute that Qian Jiaming team sets up Combining stress, CACS), this model is to give acute constraint on the basis of chronic unpredictability Stress model to answer Swash, simulation IBS patient long-term chronic stress descend because indivedual accidents aggravation situation, compared to mother-baby separation etc. other Animal model, can the more preferable many-side IBS disease such as simulation Instestinal motility and Psychology and behavior.Set up that classical acute slowly combine should Sharp IBS model (CACS)：Giving chronic unpredictability gently stress totally 21 days, and following every 7 is a cycle.At the 13rd day and After chronic unpredictability gently stress terminate in 21 days the 2nd day, then give Restraint Stress 6h.Chronic unpredictability gently stress Program:(1) 4 DEG C of ice room 15 minutes；(2) food deprivation 4 hours；(3) tilt cage 4 hours；(4) water deprivation 4 hours；(5) tide Wet bedding and padding 2 hours；(6) 12 DEG C of water temperatures are swum 5 minutes；(7) Restraint Stress 2 hours；(8) pressure tail 5 minutes；(9) food and water stripping Take 6 hours by force；(10) shock by electricity 3 minutes；(11) 12 DEG C of water temperatures are swum 10 minutes；(12) illumination is overnight；(13) moist bedding and padding 4 hours； (14) pressure tail 10 minutes；(15) Restraint Stress 3 hours；(16) moist bedding and padding 6 hours；(17) shock by electricity 6 minutes；(18) Restraint Stress 4 hours；(19) moist bedding and padding are overnight.During modeling, Normal group and model group give the physiology aqueous solution of 5%DMSO, its His group gives corresponding medicine according to packet, continuous 26 days.

2. visceral sensitivity experiment

Visceral sensitivity detects：Expansion animal Colon and rectum, can cause its stomach wall to shrink, referred to as stomach wall withdraws reflection (abdominal withdrawal reflex,AWR).AWR is the involuntary movement reflection similar with visceromotor reflex, can Reflection internal organs sensitivity, is widely used for because it is non-invasive carrying out pluck sensitivity scoring.Also take stomach wall bag The method burying electrode is detected, although but this detection can obtain objective data, due to being invasive, invasive inspection Survey, for functional disease research, we are also intended to take AWR to be scored.

Before mouse experiment, fasting can't help water 12h so as to discharge remaining feces.Mice with after ether light anaesthesia, from anal Inserting induction stimulates proctectasia (CRD) expansion catheter (6Fr, 2 mm outer diameter).After mice revival adapts to one hour, pass through Syringe gas injection increases balloon pressure, gives 0.25ml by syringe respectively, and the air of 0.35ml, 0.5ml, 0.65ml enters The air bag of catheter, respectively in triplicate, the stomach wall observing and recording mice withdraws reflection to each different pressures, is shown in Table 1.

Table 1：Mouse web portion withdraws reflection grade form

3. Instestinal motility experiment

Instestinal motility detects：Because IBS often shows, Instestinal motility is abnormal, and we adopt carbon powder method to detect Instestinal motility Situation.

Before all mice detections, fasting 24 hours.Mice, with food coloring gavage, puts to death mice after 30 minutes.Open abdomen Chamber, cuts off mesentery, stretches intestinal tube, the length of measurement pylorus to rectum and food coloring displacement.

Food coloring ink propulsive rate (%)=(distance/small intestinal total length of food coloring front end and pylorus) × 100%.System Meter credit analysis carries out statistical procedures using GraphPad Prism 6.0 statistical package, and data is represented with mean ± SEM.Just Often adopt t check analyses between matched group and stress group, medicine effect and its group difference adopt in One-wayANOVA Dunnett ' st inspection is analyzed.

2nd, result

1. the impact to IBS model mice body weight for the resveratrol

Compared with Normal group, there was no significant difference for IBS model mice body weight (P ＞ 0.05).Give resveratrol After (2.5,5 and 10mg/kg), compared with stress group, there was no significant difference for medication group Mouse Weight, is shown in Table 1 and Fig. 1.

The impact to IBS model mice body weight for table 1 resveratrol

Control

Stress

REV2.5

REV5

REV10

Rol

Despramin

Diazepam

33±0.7977

30.4±1.067

31.45±0.6791

32.45±0.5934

32.18±0.9612

32.27±0.6754

32.82±0.872

32.09±0.8029

Fig. 1 is the impact to IBS model mice body weight for the resveratrol, compared with Normal group, IBS model mice body weight There was no significant difference.IBS mice after resveratrol process, improved by body weight, but no difference of science of statistics.Rolipram, go What first imipramine and diazepam were processed stress be after mice administration, and its body weight is relative to IBS model mouse there was no significant difference (P ＞ 0.05).N=12, Mean ± SEM.

2. the impact to IBS model mice visceral sensitivity for the resveratrol

Result shows, compared with Normal group, the intestinal sensitivity of IBS model mice is giving 0.5ml and 0.65ml (P ＜ 0.0001, P ＜ 0.0001) is significantly raised during air.And giving IBS model mice resveratrol (2.5,5 and 10mg/ Kg, after), the visceral sensitivity of mice reduces.And when giving 0.35m, 0.5ml, 0.65ml air detection, resveratrol is each The effect of group significantly reduces the visceral sensitivity of IBS model mice.

The impact to IBS model mice visceral sensitivity for table 2. resveratrol

gas vol(ml)	Control	Stress	REV2.5	REV5	REV10
						0.25	0.3333±0.1231	0.6061±0.1609	0.3889±0.06901	0.3889±0.0902	0.3889±0.2203
0.35	1.083±0.0131	1.455±0.1369	0.9444±0.0803^*	0.9722±0.0643	1±0.1925
						0.5	1.722±0.0991	2.545±0.1293^####	1.722±0.135^***	1.833±0.0962^**	1.889±0.1319^*
0.65	2.389±0.0902	3.818±0.0525^####	3.417±0.1095	3±0.1589^***	3.056±0.0803^**

The impact (Continued) to IBS model mice visceral sensitivity for table 2. resveratrol

gas vol(ml)	Rol	Despramin	Diazepam
				0.25	0.5278±0.1714	0.1667±0.0649	0.3333±0.07107
0.35	1.139±0.215	0.75±0.1015^**	0.8611±0.0643^*
				0.5	2.472±0.2301	1.889±0.1553^*	1.778±0.06268^**
0.65	3.194±0.207^*	2.667±0.1692^****	2.75±0.117^****

Fig. 2 be the impact to IBS model mice visceral sensitivity for the resveratrol, A, B, C, D represent respectively give 0.25ml, During 0.35ml, 0.5ml, 0.65ml air, the intestinal sensitivity of mice.N=12, Mean ± SEM.^***P ＜ 0.001vs non- stressed vehicle group；^#P ＜ 0.05,^###P ＜ 0.001vs vehicle-treated stressed group.

3. the impact to IBS model mice Instestinal motility for the resveratrol

Result shows, compared with Normal group, in IBS model edible color food coloring ink propulsive rate significantly reduce (^## P ＜ 0.01).After giving resveratrol 23 days, the 5 IBS model mices processing with 10mg/kg resveratrol compare IBS model Mice, its food coloring ink propulsive rate dramatically increases (P ＜ 0.01, P ＜ 0.001).Through rolipram, desmethylimipramine, After process is dissolved in west, it dramatically increases (P ＜ 0.001, P ＜ 0.05, P ＜ 0.0001) using pigment ink propulsive rate.

The impact to IBS model mice food coloring ink propulsive rate for table 3. resveratrol

Control

Stress

REV2.5

REV5

REV10

Rol

Despramin

Diazepam

62.95±3.681

47.31±3.101^##

52.6±5.979

68.04±3.919^**

72.72±3.963^***

70.38±1.233^***

65.46±3.823^*

71.55±4.29^****

Fig. 3 is the impact to IBS model mice food coloring ink propulsive rate for the resveratrol, n=12, Mean ± SEM.^##P ＜ 0.01vs non-stressed vehicle group；^*P ＜ 0.05,^**P ＜ 0.01,^***P ＜ 0.001,^****P ＜ 0.0001vs vehicle-treated stressed group.

3rd, study brief summary

The studies above shows, in terms of visceral sensitivity, gives slightly significantly to raise after large quantity of air in intestinal, high, normal, basic three The resveratrol of individual dosage all can reduce the intestinal hypersensitivity of IBS model mice.In terms of Instestinal motility, IBS model mice Show weakening of Instestinal motility, middle high dose resveratrol significantly reverses weakening of Instestinal motility.Illustrate that IBS mice has intestinal Road hypersensitivity and Instestinal motility slow down, and resveratrol can reverse these changes above-mentioned.This is studied as resveratrol conduct The safely and effectively medicine for the treatment of IBS provides experimental basis.

Claims

1. application in preparation treatment irritable bowel syndrome medicine for the resveratrol.