Effect of electroacupuncture of "Hegu" (LI4) and "Zusanli" (ST36) on intestinal sensitivity and motility in irritable bowel syndrome rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Hegu"(LI4) and "Zusanli"(ST36)on changes of intestinal sensitivity and colonic motility and expression of colonic 5-hydroxytryptamine 3A receptor (5-HT3AR) in irritable bowel syndrome (IBS) rats, so as to reveal its mechanism underlying improvement of IBS. METHODS: A total of 40 neonatal Wistar rats were randomly and equally divided into normal control, model, LI4 and ST36 groups (n＝10). The IBS model was induced by mother-infant separation, acetic acid enema and colorectal distension (CRD). EA (2 Hz/100 Hz, a tolerable strength) was applied to bilateral LI4 and ST36 for 20 min, once every other day for 5 times. The Bristol stool form scale was used to assess the gastrointestinal function, and the latency and number of abdominal muscular contraction waves of abdominal withdrawal reflex (AWR) were used to evaluate the intestinal sensitivity and motility respectively. The immunoactivity of 5-HT3AR of the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the score of Bristol fecal form scale, number of muscular contraction waves and expression levels of colonic 5-HT3AR in the myometrium and mucosal layers were significantly increased (P<0.01), and the latency of muscular initial contraction wave was obviously shortened in the model group relevant to the normal control group (P<0.01). After the intervention, the increased Bristol fecal form score, number of muscular contraction waves and expression levels of 5-HT3AR in the myometrium and mucosal layers as well as the decreased latency of muscular contraction were reversed in both LI4 and ST36 groups (P<0.01, P<0.05). The effect of EA of ST36 was significantly superior to those of EA-LI4 in lowering Bristol fecal scale score and 5-HT3AR expression in the muscular layer (P<0.01), but obviously inferior to those of EA-LI4 in increasing the latency of of muscular initial contraction wave and down-regulating muscular contraction waves and 5-HT3AR expression in the mucosal layer (P<0.05, P<0.01). CONCLUSION: Both EA-LI4 and EA-ST36 can significantly improve the symptoms of abdominal pain and diarrhea, but EA-LI4 is better in suppressing intestinal high sensitivity, and EA-ST36 is better in promoting intestinal motility, suggesting a specificity of effect of acupoints of different meridians.

Effect of protease activated receptor 2 on colonic motility in diabetes mice / 上海交通大学学报（医学版）

Objective • To observe the effect of protease activated receptor 2 (PAR2) on the colonic motility in diabetic mice and investigate the mechanism. Methods • The mouse model of type 1 diabetes mellitus was established by intraperitoneal injection of streptozotocin. The smooth muscle strips and segments of colons were isolated. The effects of PAR2 agonist on colonic motility were observed by muscle strip tension contraction and colonic migrating motor complex experiments. The effect of small conductance calcium-activated potassium channel (SK3 channel) antagonist on it was also observed. Results • PAR2 agonist inhibited colonic motility and colonic smooth muscle was more sensitive to PAR2 agonist in diabetic mice. PAR2 agonist-induced inhibition was inhibited by SK3 channel antagonist. Conclusion • PAR2 activity in diabetic mice colons is significantly enhanced, which may inhibit colonic motility through SK3 channel.

Fluoroscopic Characterization of Colonic Dysmotility Associated to Opioid and Cannabinoid Agonists in Conscious Rats

BACKGROUND/AIMS: Gastrointestinal adverse effects have a major impact on health and quality of life in analgesics users. Non-invasive methods to study gastrointestinal motility are of high interest. Fluoroscopy has been previously used to study gastrointestinal motility in small experimental animals, but they were generally anesthetized and anesthesia itself may alter motility. In this study, our aim is to determine, in conscious rats, the effect of increasing doses of 2 opioid (morphine and loperamide) and 1 cannabinoid (WIN 55,212-2) agonists on colonic motility using fluoroscopic recordings and spatio-temporal maps. METHODS: Male Wistar rats received barium sulfate intragastrically, 20–22 hours before fluoroscopy, so that stained fecal pellets could be seen at the time of recording. Animals received an intraperitoneal administration of morphine, loperamide, or WIN 55,212-2 (at 0.1, 1, 5, or 10 mg/kg) or their corresponding vehicles (saline, Cremophor, and Tocrisolve, respectively), 30 minutes before fluoroscopy. Rats were conscious and placed within movement-restrainers for the length of fluoroscopic recordings (120 seconds). Spatio-temporal maps were built, and different parameters were analyzed from the fluoroscopic recordings in a blinded fashion to evaluate colonic propulsion of endogenous fecal pellets. RESULTS: The analgesic drugs inhibited propulsion of endogenous fecal pellets in a dose-dependent manner. CONCLUSIONS: Fluoroscopy allows studying colonic propulsion of endogenous fecal pellets in conscious rats. Our method may be applied to the noninvasive study of the effect of different drug treatments and pathologies.

Prediction of Delayed Colonic Transit Using Bristol Stool Form and Stool Frequency in Eastern Constipated Patients: A Difference From the West

BACKGROUND/AIMS: The correlation between the Bristol stool form scale (BSFS) and colonic transit time (CTT) has been reported in Western populations. Our study aims to study the relationship between BSFS, stool frequency, and CTT in Eastern patients with chronic constipation. METHODS: A total of 144 chronic functional constipation patients underwent colonic transit study by using radio-opaque markers, anorectal manometry, and balloon expulsion test. Stool diary including stool forms and frequency was recorded. Delayed CTT was defined as the retention of more than 20.0% of radio-opaque markers in the colon on day 5. RESULTS: Twenty-five patients (17.4%) had delayed colonic transit. Mean 5-day BSFS (OR, 0.51; 95% CI, 0.34–0.79; P = 0.021) and stool frequency (OR, 0.60; 95% CI, 0.44–0.83; P = 0.002) were independently associated with delayed CTT by logistic regression analysis. Mean 5-day BSFS (area under the curve [AUC], 0.73; 95% CI, 0.62–0.84; P < 0.001) and stool frequency (AUC, 0.75; 95% CI, 0.63–0.87; P < 0.001) fairly predicted delayed CTT. The optimal mean 5-day BSFS of ≤ 3 provided 68.0% sensitivity, 69.7% specificity, and 69.4% accuracy, and the optimal stool frequency ≤ 2 bowel movements in 5 days provided 64.0% sensitivity, 83.1% specificity, and 84.0% accuracy for predicting delayed CTT. CONCLUSIONS: Both stool form and frequency were significantly associated with delayed CTT. Stool frequency ≤ 2 and BSFS 1–3 rather than BSFS 1–2 that was used in the Westerners could be used as surrogate for delayed CTT in Eastern patients with constipation.

Role of purinergic signaling mediated by ATP in Alzheimer’ s disease-as-sociated colonic motility disorder / 中国病理生理杂志

AIM: To explore the role of purinergic signaling mediated by ATP in the Alzheimer ’ s disease (AD)-related colon motility disorder and its related molecular mechanisms .METHODS:(1)Clinical trials:AD patients in our hospital were collected and studied .Radioimmunoassay was used for the determination of plasma motilin (MTL), cholecystokinin (CCK), vasoactive intestinal peptide (VIP) and nitric oxide (NO), and high-performance liquid chroma-tography ( HPLC) was applied to test the level of adenosine triphosphate ( ATP) .The patients were assessed by neuropsy-chology and scored accordingly .( 2 ) In animal experiments , AD mice received Morris water maze test , and the spatial learning and memory function were evaluated .The plasma levels of MTL , CCK, VIP and NO were examined by radioimmu-noassay , and the level of ATP was measured by HPLC .Choline acetyltransferase ( ChAT ) , VIP, nitric oxide synthase ( NOS) and ATP synthase were detected by immunohistochemistry .Western blot and immunohistochemistry were used to detect the expression of P2Y receptor.(3) In vitro, organ bath was applied to observe the effect of α,β-methylene ATP (α,β-MeATP), an agonist of P2Y receptor, on both spontaneous and electrically evoked contraction of colonic smooth muscle strip, and the technique of intracellular microelectrode was applied to observe the effect of α,β-MeATP on the membrane potential of colonic smooth muscle cells .RESULTS:Compared with control group , the levels of MTL and CCK were decreased (P<0.01), and the levels of NO and ATP were increased (P<0.05 or P<0.01), while the VIP level was not changed.Mini-Mental State Examination (MMSE) score was decreased (P<0.05), Alzheimer’s Disease Assess-ment Scale-Cognitive Subscale (ADAS-Cog) score, Neuropsychiatric Inventory (NPI) score and Alzheimer’s Disease Co-operative Study-Activities of Daily Living Scale ( ADCS-ADL ) were all increased as compared with control group ( P <0.01).The 4~6 d escape latency of APP/PS1 AD mice was significantly prolonged (P<0.05), and the space explora-tion ability distinctly reduced (P<0.05).In AD mice, the levels of MTL and CCK were decreased (P<0.01), and the levels of NO and ATP were increased (P<0.05 or P<0.01), while the VIP level was not changed .The protein expres-sion of colonic ATP synthase was significantly increased (P<0.05), but the expression of ChAT, VIP and NOS was not changed.The expression of P2Y receptor was increased (P<0.01).The results of in vitro experiment displayed that α,β-MeATP, from 20 μmol/L to 100 μmol/L, inhibited the spontaneous contraction of colonic smooth muscle strip in the nor-mal mice and AD mice ( P<0.05 or P<0.01 ) , and this inhibition was reversed by Na +channel inhibitor tetrodotoxin (TTX) (P<0.05 or P<0.01).In addition, the effect of α,β-MeATP at 100μmol/L on the AD mice was more obvious than that on the normal mice (P<0.05), and this inhibition was also antagonized by TTX (P<0.05 or P<0.01), pro-minent in AD group as compared with control group (P<0.05).In 10 Hz electrically evoked contraction of colonic smooth muscle strip,α,β-MeATP inhibited both the normal and AD mice (P<0.05 or P<0.01), while the inhibition was more obvious in the AD mice at the concentration of 40μmol/L or 100μmol/L (P<0.05 or P<0.01).CONCLUSION:AD patients and AD mice are accompanied by decreased MTL and CCK levels , and enhanced NO level , thus inducing colonic motor dysfunction along with AD .Meanwhile, ATP in plasma, purinergic neurons , and P2Y receptor expression are in-creased in the AD mice .Purinergic signaling mediated by ATP inhibits colonic smooth muscle strip contraction and further paralyzes the colonic movement function in AD .

Neurotensin Changes Propulsive Activity into a Segmental Motor Pattern in the Rat Colon

BACKGROUND/AIMS: Neurotensin is a gut-brain peptide with both inhibitory and excitatory actions on the colonic musculature; our objective was to understand the implications of this for motor patterns occurring in the intact colon of the rat. METHODS: The effects of neurotensin with concentrations ranging from 0.1-100 nM were studied in the intact rat colon in vitro, by investigating spatio-temporal maps created from video recordings of colonic motility before and after neurotensin. RESULTS: Low concentration of neurotensin (0.1-1 nM) inhibited propagating long distance contractions and rhythmic propagating motor complexes; in its place a slow propagating rhythmic segmental motor pattern developed. The neurotensin receptor 1 antagonist SR-48692 prevented the development of the segmental motor pattern. Higher concentrations of neurotensin (10 nM and 100 nM) were capable of restoring long distance contraction activity and inhibiting the segmental activity. The slow propagating segmental contraction showed a rhythmic contraction—relaxation cycle at the slow wave frequency originating from the interstitial cells of Cajal associated with the myenteric plexus pacemaker. High concentrations given without prior additions of low concentrations did not evoke the segmental motor pattern. These actions occurred when neurotensin was given in the bath solution or intraluminally. The segmental motor pattern evoked by neurotensin was inhibited by the neural conduction blocker lidocaine. CONCLUSIONS: Neurotensin (0.1-1 nM) inhibits the dominant propulsive motor patterns of the colon and a distinct motor pattern of rhythmic slow propagating segmental contractions develops. This motor pattern has the hallmarks of haustral boundary contractions.

Barium-based Colonic Transit Detection and Individualized Treatment for Chronic Constipation / 胃肠病学

Background:Chronic constipation is a major cause of impaired quality of life in modern society. Reasonable and effective management of chronic constipation could be achieved based on the principle of evidence-based medicine and the modern concept of constipation,and this is a challenge facing the clinicians. Aims:To investigate the role of barium-based colonic transit detection in diagnosis and treatment of chronic constipation. Methods:Fifty patients with chronic constipation from Apr. 2013 to Oct. 2014 at the First Hospital of Shanxi Medical University were recruited and randomly allocated into two groups,control group and individualized treatment group. Patients in individualized treatment group received 20 barium markers orally and abdominal plain radiography was performed 48 and 72 hours later,respectively for calculating the colonic transit index. According to the type of colonic transition and the characteristics of colonic motility estimated by colonic transit index and clinical manifestations,an individualized therapeutic regimen was formulated and the efficacy was evaluated. Patients in control group were treated empirically according to the clinical manifestations. Results:Mosapride and lactulose or polyethylene glycol were administered orally in control group;when abdominal pain or abdominal distension was predominant,pinaverium bromide or trimebutine was used respectively instead of mosapride. Barium-based colonic transit detection revealed that 9 patients in individualized treatment group were slow transit constipation,6 were outlet obstructive constipation and 8 were the mixed type. After 2 weeks of empirical or individualized treatment,the defecation rates of the two groups were 24. 0%(6 / 25)and 52. 2%(12 / 23)within 24 hours and 64. 0%(16 / 25)and 87. 0%(20 / 23)within 48 hours,respectively(P all < 0. 05). Conclusions:Barium-based colonic transit detection is a simple,economical and practical modality for guiding the individualized treatment in patients with chronic constipation.

Study on colonic H2 S-producing enzyme CSE and colonic motility in diabetes mellitus / 重庆医学

Objective To investigate the relationsheep between colonic H2 S‐producing enzyme cystathionine‐γ‐lyase(CSE) and colonic motility in a rat model of diabetes mellitus(DM) .Methods To obtained diabetic rat models ,all rats were injected intra‐peritoneally (ip) 1% streptozotocin (STZ ,65 mg/kg) ,type 1 diabetes model was established ,nornmol conditions were observed pe‐riodically .Ten days after treatment ,production of colonic longitudinal smooth muscle strips ,organ bath recordings were used to test the colonic motility ;immunofluorescence and double immunofluorescence lableling method were performed to detect the distribution of CSE;Western blot were performed on rat colonic samples devoid of mucosa and submucosa to detect the expression of CSE .Re‐sults DM rats decreased the colonic motility(P0 .05);DM rats increased the expression of CSE in the colon devoid of mucosa and sub‐mucosa(P<0 .05) .Conclusion The decrease of colonic motility in diabetic rat may be associated with the increased production of H2 S‐producing enzyme CSE .

Role of Corticotropin-releasing Factor Signaling in Stress-related Alterations of Colonic Motility and Hyperalgesia

The corticotropin-releasing factor (CRF) signaling systems encompass CRF and the structurally related peptide urocortin (Ucn) 1, 2, and 3 along with 2 G-protein coupled receptors, CRF1 and CRF2. CRF binds with high and moderate affinity to CRF1 and CRF2 receptors, respectively while Ucn1 is a high-affinity agonist at both receptors, and Ucn2 and Ucn3 are selective CRF2 agonists. The CRF systems are expressed in both the brain and the colon at the gene and protein levels. Experimental studies established that the activation of CRF1 pathway in the brain or the colon recaptures cardinal features of diarrhea predominant irritable bowel syndrome (IBS) (stimulation of colonic motility, activation of mast cells and serotonin, defecation/watery diarrhea, and visceral hyperalgesia). Conversely, selective CRF1 antagonists or CRF1/CRF2 antagonists, abolished or reduced exogenous CRF and stress-induced stimulation of colonic motility, defecation, diarrhea and colonic mast cell activation and visceral hyperalgesia to colorectal distention. By contrast, the CRF2 signaling in the colon dampened the CRF1 mediated stimulation of colonic motor function and visceral hyperalgesia. These data provide a conceptual framework that sustained activation of the CRF1 system at central and/or peripheral sites may be one of the underlying basis of IBS-diarrhea symptoms. While targeting these mechanisms by CRF1 antagonists provided a relevant novel therapeutic venue, so far these promising preclinical data have not translated into therapeutic use of CRF1 antagonists. Whether the existing or newly developed CRF1 antagonists will progress to therapeutic benefits for stress-sensitive diseases including IBS for a subset of patients is still a work in progress.

Research Progress of Therapy for Chronic Constipation / 胃肠病学

Chronic constipation is a common clinical disorder,which has different influences on patients’quality of life and work. With the increase in pressure and pace of modern life,the aging of population,the changes in dietary pattern and the challenge of social psychological problems,the overall prevalence of chronic constipation in general population is increasing in recent decades. However,the therapeutic efficacy of chronic constipation is still not satisfactory. This article reviewed the advances in study on treatment of chronic constipation.

Autonomic Nerve Regulation of Colonic Peristalsis in Guinea Pigs

BACKGROUND/AIMS: Colonic peristalsis is mainly regulated via intrinsic neurons in guinea pigs. However, autonomic regulation of colonic motility is poorly understood. We explored a guinea pig model for the study of extrinsic nerve effects on the distal colon. METHODS: Guinea pigs were sacrificed, their distal colons isolated, preserving pelvic nerves (PN) and inferior mesenteric ganglia (IMG), and placed in a tissue bath. Fecal pellet propagation was conducted during PN and IMG stimulation at 10 Hz, 0.5 ms and 5 V. Distal colon was connected to a closed circuit system, and colonic motor responses were measured during PN and IMG stimulation. RESULTS: PN stimulation increased pellet velocity to 24.6 +/- 0.7 mm/sec (n = 20), while IMG stimulation decreased it to 2.0 +/- 0.2 mm/sec (n = 12), compared to controls (13.0 +/- 0.7 mm/sec, P < 0.01). In closed circuit experiments, PN stimulation increased the intraluminal pressure, which was abolished by atropine (10(-6) M) and hexamethonium (10(-4) M). PN stimulation reduced the incidence of non-coordinated contractions induced by NG-nitro-L-arginine methyl ester (L-NAME; 10(-4) M). IMG stimulation attenuated intraluminal pressure increase, which was partially reversed by alpha-2 adrenoceptor antagonist (yohimbine; 10(-6) M). CONCLUSIONS: PN and IMG input determine speed of pellet progression and peristaltic reflex of the guinea pig distal colon. The stimulatory effects of PN involve nicotinic, muscarinic and nitrergic pathways. The inhibitory effects of IMG stimulation involve alpha-2 adrenoceptors.

Effect of Pinaverium Bromide on different layers and segments of colonic smooth muscle strips in wrap restraint stress rats / 中国临床药理学与治疗学

AIM: To evaluate effects of Pinaverium Bromide on different segments and layers of colonic smooth muscle in wrap restraint stress (WRS) rats and explore its possible therapeutic mechanism on different types of irritable bowel syndrome (IBS). METHODS: Adult SD rats were randomly divided into model group (wrap restraint stress group) and control group. Colonic smooth muscle strips were made from different segments and layers in two groups. The spontaneous contraction activities of colonic longitudinal/circular muscle (LM/CM) strips of rats were observed with organ bath system before and after addition of series concentrations of pinaverium. RESULTS: Pinaverium Bromide caused concentration-dependent inhibition of colonic smooth muscle, the inhibitory effect of pinaverium in model group was significantly stronger than that in control group(proximal colon: 28.54±4.82 vs 7.48±1.65,21.75±1.00 vs 12.56±3.15; distal colon: 15.71±5.27 vs 3.89±1.16, 20.16±3.16 vs 7.56±1.96 )(P<0.05). Compared with that of distal colon, inhibitory effect of pinaverium was significantly higher of proximal colon (P<0.05). For the inhibition of pinaverium, there was no significant difference between LM and CM strips in the same intestinal segments (P>0.05). CONCLUSION: Effects of Pinaverium Bromide on different colonic muscle layers and segments in WRS rats is probably related with its therapeutic mechanism on different types of IBS.

Effect and mechanism of Ghrelin on the colonic motility of diabetic mice / 中国现代普通外科进展

Objective:To investigate colonic motility effect and mechanism of Ghrelin in diabetic mice.Methods:In vivo,the effects of Ghrelin(50,100,200 ?g/kg)on colonic transit of diabetic mice were measured by charcoal suspension pushing test.The effects of Atropine,N?-nitro-L-arginine methylester hydrochloride(L-NAME)and D-Lys3-GHRP-6(GHS-R antagonist)on the colonic transit of Ghrelin(200 ?g/kg)were also investigated.In vitro,the effects of Ghrelin on spontaneous contraction of proximal colonic circular muscle strips of diabetic mice were studied.Results:Ghrelin accelerated colonic transit of diabetic mice with significant dose-response relationship(P

Assessment of Colonic Motility and Nutrients Intake in Adult with Cerebral Palsy

OBJECTIVE: To evaluate the colonic motility and nutrients intake in adults with cerebral palsy (CP) and to compare the results with those of normal adults. METHOD: Thirty-nine adults with CP were participated. They took the radioopaque markers for 3 successive days. Then, abdominal X-ray was taken on the fourth day. The total and segmental colon transits were estimated by the simplified assessment of a single-film technique by Metcalf et al. The amounts of nutrients intake for 3 days were recorded and nutritional factors were analyzed by ESHA(r) Food Processor. Then, daily intakes of the nutrients were compared with Recommended Dietary Allowance of Korean Nutrition Society. RESULT: Colon transit time were more than 2 times delayed in adults with CP as compared with those of normal controls. Colon transit time was significantly prolonged in non- functional ambulators (p<0.05). Colon transit time tended to be delayed in subjects with spastic type, but not related with the degree of cognitive function. On the evaluation of daily nutrients intake, several nutritional factors were inadequate in adults with CP. CONCLUSION: Adults with CP had the problems in colonic motility and nutritional intake. Also, delayed colon transit time was significantly related with poor mobility, and possibly related with spasticity.

THE COLONIC TRANSIT TEST IN THE ASSESSMENT OF CHRONIC CONSTIPATION / 药物分析学报

In order to assess colonic motility of chronic constipation, colonic transit test was carried out in 34 patients with chronic constipation and in 20 healthy subjects. 20 radiopaque markers are ingested at 8 am before the day test, and plain abdominal films were obtained at 24 h, 48 h and 72 h. The normal value of colonic transit test was 16(80%), or more markers passed after 72 h. By means of transit time study, 34 constipated patients were classified into 2 groups: 12 normal transit patients and 22 slow transit patients. There was no difference in colonic transit time between normal transit patients and controls (P＞0.05). Patients with slow transit had more markers left in right colon, left colon and rectosigmoid colon at 48 h (P＜0.01, respectively) and 72 h (P＜0.01, respectively). According to the transit index, 22 slow transit patients were divided into 3 types: 10 cases colonic stasis, 8 cases outlet obstruction and 4 cases colorectal stasis. The study suggests that chronic constipated patients have abnormalbilities of colonic transit.