Effect of acupuncture and moxibustion on intestinal flora in the rats with diarrhea-predominant irritable bowel syndrome based on 16S rDNA technique / 中国针灸

OBJECTIVES@#To explore the effect of acupuncture and moxibustion on intestinal flora in the rats with diarrhea-predominant irritable bowel syndrome (IBS-D) based on 16S rDNA technique.@*METHODS@#Ten rats were randomized from 58 SPF-grade male SD rats to be the blank group. The remained 48 rats were prepared to be IBS-D models by the modified method of acetic acid enema combined with binding tail-clip stress. Forty successfully-modeled rats were randomly divided into a model group, an acupuncture group, a moxibustion group and a western medication group, with 10 rats in each one. In the acupuncture group, the needle was inserted at bilateral "Zusanli" (ST 36) and remained for 15 min in each rat. In the moxibustion group, the suspending moxibustion was delivered at bilateral "Zusanli" (ST 36) for 15 min. The rats in the western medication group were given pinaverium bromide suspension (10 mL/kg) by intragastric administration. The above interventions were performed once daily for consecutive 14 days. The body mass and the score of fecal trait were compared before and after modeling, as well as after intervention in each group. Fecal water content, diarrhea index and colon transit time (CTT) were measured after modeling and intervention in the rats of each group separately. After intervention, the colonic morphology of rats in each group was observed, and using 16S rDNA technique, the intestinal flora was detected.@*RESULTS@#After modeling, compared with the blank group, the body mass and CTT were reduced (P<0.01); fecal trait scores, fecal water contents and diarrhea index increased (P<0.01) in the other 4 groups. After intervention, the body mass and CTT of the rats decreased (P<0.01), and fecal trait score, fecal water content and diarrhea index increased (P<0.01) in the model group compared with those in the blank group. In the acupuncture group, the moxibustion group and the western medication group, when compared with the model group, the body mass and CTT were elevated (P<0.01), while fecal trait scores, fecal water contents and diarrhea index declined (P<0.01). Compared with the western medication group, fecal water content decreased in the acupuncture group and the moxibustion group (P<0.05), while CTT increased in the acupuncture group (P<0.01), the body mass increased and fecal trait score was dropped in the moxibustion group (P<0.05). The colonic mucosa structure was clear and complete, and there was no obvious inflammatory cell infiltration in the blank group. The mild interstitial edema of intestinal mucosa was presented with the infiltration of few inflammatory cells in the model group. There was the infiltration of few inflammatory cells in the mucosa of the acupuncture group, the moxibustion group and the western medication group. Compared with the blank group, the indexes of Richness, Chao1, ACE and Shannon decreased in the model group (P<0.05). Indexes of Richness, Chao1 and ACE increased in the acupuncture group and the moxibustion group (P<0.05), and the Richness index in the western medication group increased (P<0.05) when compared with those in the model group. The relative abundance of Bacteroidetes, Proteobacteria and Prevotella increased (P<0.05), and that of Firmicutes and Muribaculaceae decreased (P<0.05) in the model group compared with those in the blank group. When compared with the model group, the relative abundance of Bacteroidetes, Proteobacteria and Prevotella was reduced (P<0.05), while that of Firmicutes and Muribaculaceae increased (P<0.05) in the acupuncture group, the moxibustion group and the western medication group; and that of Actinobacteria and Bifidobacterium increased in the acupuncture group and the moxibustion group (P<0.05). Compared with the blank group, the relative abundance of lipopolysaccharide (LPS) biosynthesis was elevated (P<0.05), and that of folate biosynthesis, lipoic acid metabolism, zeatin biosynthesis, ubiquinone and other terpenoid quinone biosynthesis decreased (P<0.05) in the model group. The relative abundance of LPS biosynthesis was dropped (P<0.05), and that of folate biosynthesis, lipoic acid metabolism, zeatin biosynthesis, ubiquinone and other terpenoid quinone biosynthesis increased (P<0.05) in the acupuncture group, the moxibustion group and the western medication group compared with those of the model group.@*CONCLUSIONS@#Either acupuncture or moxibustion can relieve the symptoms of IBS-D and protect intestinal mucosa, which may be associated with regulating the structure of intestinal flora and promoting nutrient metabolism and biosynthesis.

Hunyuan moxibustion for diarrhea-predominant irritable bowel syndrome of spleen and kidney yang deficiency: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the therapeutic effect between Hunyuan moxibustion and oral western medication on diarrhea-predominant irritable bowel syndrome（IBS-D）of spleen and kidney yang deficiency.@*METHODS@#Sixty patients with IBS-D of spleen and kidney yang deficiency were randomly divided into a Hunyuan moxibustion group and a western medication group, 30 cases each group. The Hunyuan moxibustion group was treated with Hunyuan moxibustion at Guanyuan（CV 4），40 min each time, once a day; in the western medication group，loperamide hydrochloride capsules (2 mg each time, 3 times a day) and bacillus licheniformis live capsules (0.5 g each time, 3 times a day) were given orally.Both groups were treated for 20 days. The scores of irritable bowel syndrome（IBS）symptom severity scale（IBS-SSS）, IBS quality of life scale (IBS-QOL) and TCM symptom grading quantitative were observed before and after treatment, and the clinical efficacy and safety were evaluated in the two groups.@*RESULTS@#After treatment，each item scores and total scores of IBS-SSS in the two groups were lower than those before treatment（P<0.05）, and the total scores of IBS-QOL were higher than those before treatment (P<0.05)；each item score and total score of IBS-SSS in the Hunyuan moxibustion group were lower than those in the western medication group (P<0.05), and the total score of IBS-QOL in the Hunyuan moxibustion group was higher than that in the western medication group (P<0.05).After treatment, each item score and total score of TCM symptom grading quantitative in the Hunyuan moxibustion group were lower than those before treatment (P<0.05), the abdominal pain, diarrhea, lack of appetite scores and total score in the western medication group were lower than those before treatment (P<0.05)；and the abdominal pain, soreness and weakness of waist and knees, fear to cold and cold limbs scores and total score in the Hunyuan moxibustion group were lower than those in the western medication group (P<0.05).The total effective rate was 90.0%（27/30）in the Hunyuan moxibustion group, which was higher than 73.3%（22/30）in the western medication group (P<0.05). No adverse reactions occurred in both groups during treatment.@*CONCLUSION@#Hunyuan moxibustion can effectively improve the symptom severity and quality of life in patients with IBS-D of spleen and kidney yang deficiency, especially in improving the symptoms of abdominal pain, soreness and weakness of waist and knees, fear to cold and cold limbs.Its therapeutic effect is superior to western medication.

Linderae Radix water extract treats diarrhea-predominant irritable bowel syndrome in rats: a serum metabolomics study / 中国中药杂志

This study aims to investigate the mechanism of Linderae Radix water extract(LRWE) in the prevention and treatment of diarrhea-predominant irritable bowel syndrome(IBS-D) based on serum metabolomics. Eighteen 2-week-old male SD rats were randomized into control, IBS-D model, and LRWE groups. The rats in other groups except the control group received gavage of senna concentrate combined with restraint stress for the modeling of IBS-D. The rats in the LRWE group were administrated with LRWE(5.4 g·kg~(-1)) by gavage, and those in the control and IBS-D model groups with an equal volume of distilled water for a total of 14 days. The visceral sensitivity was evaluated by the abdominal withdrawal reflex(AWR) score, and the degree of diarrhea was assessed by the fecal water content(FWC). The morphological changes of the colon and the morphology and number of goblet cells were observed by hematoxylin-eosin(HE) and periodic acid-schiff(PAS) staining, respectively. Ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was used for the screening of the potential biomarkers in the rat serum and their related metabolic pathways. The results showed that LRWE reduced the AWR score, decreased FWC, and alleviated visceral sensitivity and diarrhea symptoms in IBS-D rats. HE and PAS staining showed that LRWE mitigated low-grade intestinal inflammation and increased the number of mature secretory goblet cells in the colonic epithelium of IBS-D rats. A total of 25 potential biomarkers of LRWE in treating IBS-D were screened out in this study, which were mainly involved in riboflavin, tryptophan, glycine, serine and threonine metabolism, glyoxylate and dicarboxylate metabolism, and cysteine and methionine metabolism. The regulatory effects were the most significant on the riboflavin and tryptophan metabolism pathways. LRWE may alleviate the visceral hypersensitivity by promoting energy metabolism and amino acid metabolism, enhancing intestinal barrier function, and improving intestinal immune function in IBS-D rats.

Syndrome Differentiation-based Treatment of Diarrhea-predominant Irritable Bowel Syndrome with Chinese Medicine via 5-HT Signaling Pathway： A Review / 中国实验方剂学杂志

Diarrhea-predominant irritable bowel syndrome （IBS-D） is a chronic functional bowel disorder characterized by abdominal pain and diarrhea， with visceral hypersensitivity and abnormal gastrointestinal dynamics as the pathophysiological basis. The brain-gut interaction plays a role in pain-related functional gastrointestinal disorders， especially IBS-D. 5-Hydroxytryptamine （5-HT）， as an important brain-gut peptide regulating gastrointestinal function， affects brain activity， gastrointestinal motility， pain perception， mucosal inflammation， and immune response through brain-gut interaction and is associated with the occurrence and development of IBS-D. In recent years， traditional Chinese medicine （TCM） has shown great potential to mitigate gastrointestinal symptoms and improve the quality of life with its holistic view and treatment based on syndrome differentiation. Studies have shown that TCM treats IBS-D by regulating the 5-HT signaling pathway. With a focus on syndrome differentiation in TCM， this paper systematically describes the efficacy and mechanism of TCM in treating different TCM syndromes of IBS-D via the 5-HT signaling pathway， aiming to provide a scientific basis for TCM treatment of this disease.

Clinical Efficacy of Tongxie Yaofang on Diarrhea-predominant Irritable Bowel Syndrome （IBS-D） Patients with Liver Depression and Spleen Deficiency / 中国实验方剂学杂志

ObjectiveTo explore the clinical efficacy and mechanism of Tongxie Yaofang in treating diarrhea-predominant irritable bowel syndrome（IBS-D） patients with liver depression and spleen deficiency. MethodA total of 168 IBS-D patients with liver depression and spleen deficiency who were treated from August 2017 to June 2021 were divided into observation group and control group by random number table，84 in each group. The observation group was administrated with Tongxie Yaofang decoction-free granules orally，and the control group received oral treatment of pinaverium bromide，both for 4 weeks. The main symptoms of IBS were compared before and after treatment，such as the degree of abdominal pain，stool changes，traditional Chinese medicine pattern curative effect scoring system（TCM-PES），IBS quality of life questionnaire （IBS-QOL），IBS symptom severity scale（IBS-SSS），self-rating anxiety scale （SAS），and self-rating depression scale（SDS）. Nimodipine was used to evaluate the efficacy based on TCM syndrome score of liver depression and Qi stagnation. Enzyme-linked immunosorbent assay（ELISA） was conducted to detect the plasma interleukin-10（IL-10）and IL-12 before and after treatment. ResultAfter 4 weeks of treatment， the response rate of abdominal pain in observation group was 92.86% （78/84）， higher than that in control group （82.14%， 69/84）（χ2=6.254，P<0.05）. The response rates of diarrhea in observation group and control group were 91.67% （77/84）and 77.38% （65/84）， respectively（χ2=8.214，P<0.01）. TCM-PES and IBS-QOL scores of observation group after treatment were higher and IBS-SSS score was lower than those of control group （P<0.05）. The efficacy rate of TCM syndromes in observation group was higher than that of control group （P<0.05）. Additionally， after treatment， the observation group had lower SAS and SDS scores （P<0.05）and IL-12 level（P<0.05）and higher plasma IL-10 level than the control group （P<0.05）. ConclusionTongxie Yaofang can relieve abdominal pain and diarrhea in IBS-D patients with liver depression and spleen deficiency，reduce negative emotion，and improve the quality of life of patients，which may be related to alleviating the visceral hypersensitivity.

Anti-inflammation effect of moxibustion for rats with diarrhea-predominant irritable bowel syndrome based on multiple miRNAs regulating NF-κB signal pathway / 中国针灸

OBJECTIVE@#To observe the effect of moxibustion on the regulation of nuclear factor-kappa B (NF-κB) and inflammatory factors by multiple microRNAs (miRNAs) in rats with diarrhea-predominant irritable bowel syndrome (IBS-D), and to explore the anti-inflammatory mechanism of moxibustion on IBS-D.@*METHODS@#Twelve of 52 newborn rats were randomly selected into a normal group. The remaining rats were made into IBS-D model. A total of 36 rats with successful model were randomly divided into a model group, a medication group and a moxibustion group, 12 rats in each group. The rats in the medication group were intraperitoneally injected with pyrrolidine dithiocarbamate (PDTC). The rats in the moxibustion group were treated with moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for 20 min each time. All the intervention was given once a day for 7 days. Before and after modeling as well as after intervention, the body mass, loose stool rate and the minimum volume threshold of abdominal withdrawal reflex (AWR) were measured. After intervention, the contents of serum tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-8 were detected by ELISA method; the morphology of colon tissues was observed by HE staining, and the expressions of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues were detected by real-time PCR. The expressions of NF-κB p65, TNF-α, IL-1β and IL-8 protein in colon tissues were detected by immunofluorescence.@*RESULTS@#After modeling, the body mass and the minimum volume threshold of AWR in the model group were lower than those in the normal group (P<0.01); the rates of loose stool in the model group were higher than those in the normal group (P<0.01); after intervention, in the model group, the inflammatory infiltration of colon tissues was obvious, and the serum levels of TNF-α, IL-1 β, IL-8 were higher than those in the normal group (P<0.05); the expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in colon tissues was higher than that in the normal group (P<0.05); the protein expression of NF-κB p65, TNF-α, IL-1β, IL-8 was also higher than that in the normal group (P<0.01). After intervention, the body mass and the minimum volume threshold of AWR in the medication group and the moxibustion group were both higher than those in the model group (P<0.05); the loose stool rate in the medication group and the moxibustion group were lower than those in model group (P<0.05); the inflammatory cells infiltration in the colon tissues was less, the serum levels of TNF-α, IL-1β and IL-8 as well as the protein expression of NF-κB p65, TNF-α, IL-1β and IL-8 in the colon tissues in the medication group and the moxibustion group were lower than those in the model group (P<0.05, P<0.01). The expression of miR-125b, miR-31, miR-18a and NF-κB p65 mRNA in the medication group were lower than those in the model group (P<0.05). The expression of miR-155, miR-125b, miR-29b, miR-31, miR-18a and NF-κB p65 mRNA in the moxibustion group were lower than those in the model group (P<0.05). The miR-155, miR-125b, miR-29b, miR-31, miR-18a were positively correlated with NF-κB p65 mRNA (0<r<1, P<0.01).@*CONCLUSION@#The anti-inflammatory mechanism of moxibustion at "Tianshu" (ST 25) and "Shangjuxu" (ST 37) for IBS-D rats may be related to regulating multiple miRNAs to inhibit NF-κB signal pathway and reduce the expression of inflammatory factors.

CHU Hao-ran's experience in treatment of diarrhea-predominant irritable bowel syndrome with acupuncture and moxibustion / 中国针灸

Professor CHU Hao-ran believes that the clinical diagnosis and treatment of diarrhea-predominant irritable bowel syndrome with acupuncture and moxibustion should concentrate on the integration of disease and syndrome differentiation. According to the evolution of pathogenesis, this disease is divided into 4 stages, i.e. invasion of exogenous pathogens, liver stagnation and spleen deficiency, spleen and kidney yang deficiency and interaction of cold and heat. Hence, the treatment should be provided by stages. Focusing on regulating the spleen and stomach, a specific acupuncture-moxibustion prescription is proposed, including Tianshu (ST 25), Zhongwan (CV 12), Zusanli (ST 36) and Shangjuxu (ST 37). Additionally, the combination of distal and nearby acupoints is considered and an unique needling manipulation, named the fight between dragon and tiger, is exerted. On the base of syndrome and meridian differentiation, the modern medicine is combined, stressing on the mind regulation and the mutual application of acupuncture and moxibustion in treatment. Eventually, a distinctive treatment system with acupuncture and moxibustion is formed for "regulating the spleen and stomach and differentiating meridians".

Diarrhea-predominant irritable bowel syndrome of spleen deficiency and damp excess treated with fire needling therapy with filiform needle and acupoint application therapy: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the clinical therapeutic effect on diarrhea-predominant irritable bowel syndrome (IBS-D) of spleen deficiency and damp excess among fire needling therapy with filiform needle combined with acupoint application therapy, simple acupoint application therapy and pinaverium bromide tablets, and explore the mechanism on the improvements in IBS-D.@*METHODS@#A total of 150 patients with IBS-D of spleen deficiency and damp excess were randomized into a combined treatment group (50 cases, 14 cases dropped off), an acupoint application group (50 cases, 16 cases dropped off ) and a western medication group (50 cases, 13 cases dropped off ). In the western medication group, pinaverium bromide tablets were prescribed for oral administration, 50 mg per time, three times a day. In the acupoint application group, the herbal plaster of @*RESULTS@#Except the score for abdominal pain in the acupoint application group and the scores for abdominal pain and abdominal distention in the western medication group after 7 days of treatment, the scores for abdominal pain, abdominal distention, defecation frequency, stool form and incomplete bowel movement after 7 and 28 days of treatment were all reduced as compared with those before treatment in each group separately (@*CONCLUSION@#The combined treatment with the fire needling therapy with filiform needle and the acupoint application therapy effectively relieves the clinical symptoms, improves the quality of life and strengthens the immunity in the patients with IBS-D. The therapeutic effect of this combined regimen is better than either simple acupoint application therapy or the oral medication of pinaverium bromide. The mechanism may be related to the regulation of the levels of T lymphocyte subsets and the modulation of the expressions of 5-HT3R and 5-HT4R mRNA in colon tissue.

Correlation between curative effect and 5-HTTLPR polymorphism in treatment of diarrhea-predominant irritable bowel syndrome with acupuncture for regulating / 中国针灸

OBJECTIVE@#To compare the curative effect on diarrhea-predominant irritable bowel syndrome (IBS-D) between acupuncture for regulating @*METHODS@#A total of 231 patients with IBS-D were randomized into an acupuncture group (154 cases) and a western medication group (77 cases) at the ratio of 2 to 1. In the acupuncture group, acupuncture was applied to acupoint regimen for regulating @*RESULTS@#After treatment and in follow-up, the total scores of IBS-SSS in the patients of the two groups were all reduced as compared with those before treatment (@*CONCLUSION@#Acupuncture for regulating

Effect of electroacupuncture on intestinal epithelial mucosal barrier function in rats with diarrhea-predominant irritable bowel syndrome / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) on intestinal epithelial mucosal barrier function in diarrhea-predominant irritable bowel syndrome(IBS-D) rats, so as to explore its mechanisms underlying improvement of IBS-D. METHODS: Forty SD rats (half male and half female) were randomly divided into control, model, EA and medication (Pinaverium Bromide, PB) groups, with 10 rats in each group. The IBS-D model was established by chronic unpredictable mild stress combined with gavage of Senna-leaf solution. EA (2 Hz/15 Hz，0.1-1 mA) was applied to unilateral "Zusanli"(ST36),"Tianshu" (ST25), "Sanyinjiao"(SP6) and "Taichong"(LR3) alternatively for 15 min, once daily for 14 days. Rats of the medication group was treated by gavage of PB (10 mL·kg-1·d-1) for 14 days. The visceral sensitivity (pain) was assessed by using the pressure threshold which the inserted rectal balloon catheter air-inflation (connected to a blood pressure gauge) induced stronger abdominal muscular contraction to force the rat's abdomen to lift the experimental stand surface. The diarrhea index was used to evaluate loose stool grade. The expression of Claudin-1 and Occludin (intestinal epithelial tight junction proteins) of colon tissue was detected by immunohistochemistry. The activity of plasma diamine oxidase (DAO) was assayed by using spectrophotometry. RESULTS: Compared with the control group, the diarrhea index and plasma DAO activity in the model group were significantly increased (P0.05). CONCLUSION: Electroacupuncture can significantly improve abdominal pain and diarrhea in IBS-D model rats, which may be closely associated with its effects in up-regulating the expression of intestinal epithelial tight junction proteins Claudin-1 and Occludin to restore the function of intestinal epithelial mucosal barrier.

Effect of Intestinal Microbiota in Regulating Visceral Hypersensitivity of IBS-D Based on Brain-gut Interacting Mechanism / 中国实验方剂学杂志

Diarrhea-predominant irritable bowel syndrome (IBS-D) is not only a biological mental disorder, but also a kind of chronic functional bowel disease induced by many factors. The pathogenesis of IBS-D is related to visceral sensory abnormalities, intestinal dynamics abnormalities, intestinal mucosal micro-inflammatory reactions, heredity, dietary intolerance and other factors. The visceral hypersensitivity is one of the main pathophysiology, and refers to an unknown cause of intestinal hypersensitivity to cold, bad mood and other stimuli. However, its mechanism remains unclear. The pathogenesis of IBS-D is closely related to the disturbance of brain and intestinal interaction. IBS-D patients often suffer from anxiety, depression and other psychiatric symptoms due to repeated illness, but long-term chronic mental stress can induce and aggravate IBS-D visceral hypersensitivity. Brain-derived neurotrophic factor (BDNF) and its high-affinity receptor tyrosine kinase B (TrkB) are hotspots in studies about brain-gut axis. BDNF is a highly expressed cytokine in the central nervous system and gastrointestinal tract, and can promote the development of the nervous system, maintain the normal function of mature nerve cells and regulate the gastrointestinal motility and visceral sensitivity. Intestinal microbiota is the key link of brain-gut interaction. Mental disorders are closely related to intestinal symptoms caused by changes in intestinal microbial environment. Repeated mental stimulation can lead to changes in intestinal flora; on the other hand, changes in intestinal flora structure are closely related to the development of the nervous system and the function of the brain. With intestinal microbiota as the study object, this article mainly discusses the effect of intestinal microbiota in regulating visceral hypersensitivity of IBS-D based on brain-gut axis (BDNF/TrkB signaling pathway).

Effect of Changji'an Prescription on Intestinal Permeability of IBS-D Rats / 中国实验方剂学杂志

Objective:To observe the effect of Changji'an prescription on intestinal permeability in diarrhea-predominant irritable bowel syndrome (IBS-D) rats and explore its mechanism for treatment of IBS-D. Method:Male SD neonatal rats were randomly divided into five groups:normal group, model group, pinaverium bromide group(0.018 g·kg-1), high-dose(33.48 g·kg-1) and low-dose (16.74 g·kg-1)Changji'an prescription groups. Except for the normal group, the IBS-D model was established by the combination of maternal and infant separation+acetic acid stimulation+restraint stress. After drug treatment, the ultrastructure of rat intestinal mucosa was observed by using transmission electron microscopy and the plasma D-lactate level was detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of tight junction proteins Occludin and Claudin-1 were detected by Western blot. The mRNA expression levels of Occludin,Claudin-1 and zonula occluden(ZO)-1 were detected by real time polymerase chain reaction (Real-time PCR). Result:As compared with the normal group, the intestinal mucosal epithelial cells were damaged in IBS-D model group, and the microvilli arrangement was sparse and tight junction was widened, and some were not obvious,and the plasma D-lactate level in IBS-D rats was increased significantly (PPD-lactate level in pinaverium bromide group and high-dose Changji'an prescription group was significantly decreased (PD-lactate level in the low-dose group Changji'an prescription group had a tendency to decrease with no statistical difference. The mRNA and protein expression levels of Occludin and Claudin-1 and the mRNA expression of ZO-1 in the colon of rats in each administration group were higher than those in the model group (PConclusion:The therapeutic effect of Changji'an prescription on IBS-D may be achieved by improving the intestinal permeability.

Exprossion of TLR4 and NF-κB in rats model induced with IBS-D and its mechanism / 解放军医学杂志

Objective: To analyze the dynamic changes of inflammatory factors in rats model induced with IBS-D. Methods: Thirty Wistar rats were randomly divided into the blank group, the IBS-D group, and the TAK-242 group. All rats received acute and chronic stress method, followed by no further treatment for the blank group. To induce IBS-D, before performing acute and chronic stress, the IBS-D group received 3 mg/kg saline. To explore the potential mechanism of TLR4 in IBS-D, TAK-242, an antagonist of TLR4 was given to 3 mg/kg TAK-242 group after they received acute and chronic stress. Fecal traits were evaluated by Bristol classification at 0, 7, 14, 21, 28 days, and the levels of MyD88, IL-1β, IL-6 were quantified by Elisa assays; and the levels of TLR4, NF-κB were detected by Western blot. Results: In the blank group, there was no significant change in the scores of Bristol stool and expression levels of MyD88, IL-1β, IL-6, TLR4 and NF-κB at the 7th, 14th, 21st and 28th day. Compared with the blank group, in the IBS-D rats and TAK-242 rats, the scores of Bristolian stools (P<0.05), the levels of MyD88 and serum inflammatory factors (P<0.05), TLR4, NF-κB were all increased (P<0.05). However, the scores of Bristolian stools and the expression levels of MyD88, IL-1β, IL-6, TLR4, and NF-κB in the TAK-242 group were lower than those in the IBS-D group, suggesting therapeutic effects of TAK-242 in IBS-D. Conclusions: IBS-D may increase inflammatory factors through activating the TLR4/MyD88/NF-κB signaling pathway, resulting in disease progression.

Improvement and Evaluation of Rat Model of Diarrhea-predominant Irritable Bowel Syndrome Induced by Chronic Restrain Stress / 广州中医药大学学报

Objective To modify the rat model of diarrhea-predominant irritable bowel syndrome(IBS-D) induced by chronic restrain stress,and to evaluate the modified model. Methods Thirty specific-pathogen free Sprague Dawley rats were randomly divided into control group,one-hour model group and 2-hour model group, 10 rats in each group and half being male. One-hour model group and 2-hour model group were given restrain stress one, 2 hour (s)per day respectively, and the treatment lasted 14 days. On day 7, 14, 21, 28 after modeling,the number of feces grains within 4 hours,area of the excrement and body mass were monitored. On modeling day 28 , abdominal withdrawal reflex (AWR) scores for visceral sensitivity were calculated , the pathological features of gastric antrum, duodenal, ileac, colonic tissues were observed by hematoxylin-eosin staining, and the expression level of 5-hydroxytryptamine-3A receptor (5-HT3AR) in rat colon tissue was detected by real time quantitative polymerase chain reaction(qPCR). Results Compared with control group,the number of feces grains in one-hour model group showed no obvious changes on day 7, 14, 21, 28, but the area of the excrement was obviously increased and body mass was decreased on day 14 (P < 0.05 or P < 0.01);in 2-hour model group,the number of feces grains and area of the excrement were increased on day 21,28, and body mass was decreased obviously on day 7,14,21,28 (P < 0.05 or P < 0.01). One-hour model group and 2-hour model group had higher AWR scores than the control group (P < 0.05),but the difference between the two model groups was insignificant(P > 0.05). The optical microscopy results showed that no various changes in gastric, duodenal, ileac, colonic tissues in the two model groups as compared with those in the control group. QPCR results showed that the expression level of 5-HT3AR in rat colonic tissues of the two model groups was higher than that of the control group (P < 0.05), but the expression level of 5-HT3AR in rat colonic tissues of model was not significantly different from that of the control group(P > 0.05). Conclusion IBS-D model can be successfully established with chronic restrain stress for 2 hours per day and lasting for 14 days. Excrement area and AWR scores can be used as the objective indexes for the evaluation of IBS-D model.

Effect of compound Lactobacillus acidophilus on serum SP, VIP, 5-HT, IL-6 and TNF-αin patients with diarrhea-predominant irritable bowel syndrome / 中国生化药物杂志

Objective To investigate the effect of compound Lactobacillus acidophilus on the levels of serum SP , VIP, 5-HT, IL-6 and TNF-αin patients with diarrhea-predominant irritable bowel syndrome ( IBS) .Methods 82 cases of patients with diarrhea -predominant irritable bowel syndrome ( IBS) from November 2015 to November 2016 in our hospital were selected,and randomly divided into two groups.30 patients in observation group were treated with compound Lactobacillus acidophilus tablets and 52 patients in control group were treated with trimebutine maleate capsule.The levels of serum SP, VIP, 5-HT, IL-6 and TNF-α,quality of life score,clinical curative effect and adverse reaction were observed and compared before and after treatment.Results After treatment,the diarrhea, abdominal distention, defecation frequency and abnormal rate of stool were significantly lower than those before treatment(P <0.05), the levels of SP,VIP,5-HT,IL-6 and TNF-αin serum were significantly lower than those before treatment(P <0.05),and the treatment group were lower than those of the control group.After treatment, the mental status, emotional control and work status scores of the observation group were higher than those of the control group,and the effective rate of the observation group 86.7%was significantly higher than the control group 61.5%,the differences between the two groups were statistically significant (P <0.05).There was no significant adverse events were observed during the treatment.Conclusion Compound Lactobacillus acidophilus is effective in the treatment of diarrhea-predominant irritable bowel syndrome.It can restore the levels of serum SP, VIP, 5-HT, IL-6 and TNF-αin patients with diarrhea-predominant irritable bowel syndrome and improve the quality of life .

Visceral sensitivity, gut barrier function and autonomic nerve function in patients with diarrhea-predominant irritable bowel syndrome / 中南大学学报(医学版)

Objective:To evaluate visceral sensitivity,gut barrier function and autonomic nerve function (ANF) in patients with diarrhea-predominant irritable bowel syndrome (IBS-D),and to explore their roles in IBS-D pathophysiology.Methods:A total of 46 IBS-D patients (IBS-D group) were selected from the Department of Gastroenterology of China-Japan Friendship Hospital from October 2015 to March 2016,and 20 healthy volunteer were served as a control group (HC group).Clinical and psychological symptoms were evaluated by questionnaire,and visceral sensitivity to rectal balloon distention,gut barrier function and autonomic nerve function (ANF) were examined.The difference in the abovementioned indexes were compared between the 2 groups,and the correlations in the parameters were analyzed in the IBS-D group.Results:The scores of IBS symptom severity scale (IBS-SSS),Hamilton anxiety scale (HAMA),Hamilton depression scale (HAMD) and visceral sensitivity index (VSI) were significantly higher in the IBS-D group than those in the HC group (P＜0.01).In the visceral sensitivity test,maximum tolerable threshold in the IBS-D group was significantly decreased compared to that in the HC group (P＜0.01);there was no significant difference in first sensation threshold and defecating sensation threshold between the two groups (P＞0.05).As gut barrier function markers,the serum diamine oxidase (DAO) and d-lactate were significantly increased in the IBS-D group (P＜0.05).In ANF test,the total score and parasympathetic score as well as the proportion of abnormal scores in the IBS-D group were significantly higher than those in the HC group (P＜0.05).In IBS-D group,the HAMA,VSI and serum DAO were positively correlated with IBS-SSS (r=0.528,0.575,0.507;P＜0.01),while the 3 visceral sensitivity thresholds were negatively correlated with IBS-SSS (r=-0.636,-0.476,-0.697;P＜0.01);in addition to the IBS-SSS,the HAMA,HAMD,VSI and serum DAO were also significant negatively correlated with the visceral sensitivity thresholds (all P＜0.05);no significant correlations were found between the ANF and the other parameters.Conclusion:IBS-D patients show psychological symptoms,visceral hypersensitivity,impaired gut barrier function and abnormal ANF characterized by parasympathetic dysfunction;the former 3 factors are all associated with disease severity,and thus may play vital roles in IBS-D pathophysiology.

Correlation between food intolerance and colon mucosal mast cells and substance P in 27 patients with diarrhea predominant irritable bowel syndrome / 中华消化杂志

Objective To investigate the significance and possible mechanism of food intolerance in diarrhea-predominant irritable bowel syndrome (IBS-D).Methods Twenty-seven IBS-D patients matched the Rome Ⅲ criteria were selected as IBS-D group.Twenty-seven healthy individuals without gastrointestinal symptoms were assigned to control group.Food intolerance situation of two groups were analyzed with food intolerance status evaluation questionnaire,and detection of 14 food specific IgG antibody.The severity of IBS symptoms of IBS-D group were scored by IBS-symptom severity scale (SSS).Two pieces of mucosal tissues of both ileocecal junction and sigmoid colon were obtained under colonoscopy.The content of mucosal substance P (SP) was determined by immunohistochemistry.The quantity of mast cells were detected by Giemsa staining.Chi square test and Fisher exact probability method were performed for rate comparison between two groups.Measurement data were compared with t test and Mann-Whitney rank sum test.The correlation between IBS-SSS score and mast cells and the expression of SP positive cells were analyzed by Spearman rank correlation analysis.Results There was no statistically significant difference in positive rates of food specific IgG between two groups (x2 =3.085,P=0.389),however according to food intolerance status evaluation questionnaire,the incidence of food intolerance of IBS-D group was 44.4 ％ (12/27) which was higher than that of control group (14.8 ％,4/ 27),the difference was statistically (x2 =5.684,P=0.017).Food intolerance severity index of IBS-D group (median:0(0,60)) was higher than that of control group (median:0(0,0)),and the difference was statistically significant (U=239.50,P =0.007).In foods that may cause intolerance,the percentage of foods rich in fermentable oligosaccharides and monosaccharides,disaccharides and polyol (FODMAP) such as milk,noodles,soybeans was up to 71.4％ (30/42).The expression tates of SP positive cells in the mucosa of ileocecal junction and sigmoid colon of patients with IBS-D were higher than those of the control group (x2-20.735 and 22.071,both P＜0.01).The numbers of mast cell in the mucosa of ileocecal junction and sigmoid colonic of patients with IBS D (2.40 ± 1.04/high power field (HPF) and 2.35±1.11/HPF) were more than those of the control group (0.97 ± 0.70/HPF and 0.89 ± 0.72/HPF),and the difference was statistically significant (t=-5.850 and-5.629,both P＜0.01).The severity of bowel symptom of patients with IBS-D was moderately correlated with the number of mast cells in the mucosa of sigmoid colon (r=0.576,P=0.002),and was moderately correlated with the expression of SP positive cells (r=0.691,P＜0.01).Conclusions There may be relation ship among low-grade inflammation of intestinal mucosa,food intolerance and severity of intestinal symptoms in patients with IBS-D.

Association between diarrhea-predominant irritable bowel syndrome and HTR3A, HTR3E gene polymorphism in Yangzhou, Jiangsu province, China / 中华流行病学杂志

Objective To investigate the relationship between diarrhea-predominant irritable bowel syndrome (D-IBS) and HTR3A,HTR3E gene polymorphism in Yangzhou,Jiangsu province.Methods Polymerase chain reaction (PCR) amplification and restriction fragment length pdymorphism (RFLP) technique were used to detect the 5′-UTR variant c.-42C＞T of HTR3A and 3′-UTR variant c.* 76G＞A of HTR3E on 300 healthy subjects and 450 patients with D-IBS.Results There was significant difference noticed between the D-IBS patients and the controls in the genotype of c.-42C＞T sites (P＜0.05),while the frequency of T allele was significantly higher in both female and male patients than that in the controls (P＜0.05).The frequencies of G/A genotype and A allele of c.*76G＞A sites were significantly higher in the female-patient group than that in the controls (P＜0.05),however,there was no significant difference between those male patients and controls (P＞0.05).When comparing to the CC genetype of rs1062613 site and GG genetype of rs62625044 site,the TT and GA genotype were closely related to the D-IBS,with the risk coefficients as 0.29 (95％ CI:0.14-0.61) and 0.62 (95％ CI:0.39-0.99),with statistically significant differences.Conclusion T allele and GA genetype might respectively serve as the predisposing factors of D-IBS and on the female D-IBS patients.

Clinical Research of Simotang Oral Liquid and Compound Glutamine Enteric Capsules for Treating Diarrhea-predominant Irritable Bowel Syndrome / 中国中医药信息杂志

Objective To observe the clinical efficacy of Simotang oral liquid combined with compound glutamine enteric capsules in the treatment of diarrhea-predominant irritable bowel syndrome (IBS-D) belonging to liver depression and spleen deficiency pattern, and to explore its possible mechanism. Methods Sixty patients were randomly divided into two groups. The treatment goup (30 cases) took Simotang oral liquid and compound glutamine enteric capsules, the control group took Cerekinon and Medilac-S. The treatment course was 4 weeks. Clinical symptoms, 5-hydroxytryptamine (5-HT) and substance P (SP) of both groups were evaluated respectively before and after treatment. Recurrence rate and adverse reactions were recorded. Results The total effective rate after treating 1, 2, 4 weeks between the two groups had no significant difference (P>0.05). Four weeks later, in the treatment group, total score of the symptoms, abdominal distension and defecation urgency were apparently lower than that of control group (P0.05). Obvious side effects were not found in the two groups. Conclusion Simotang oral liquid combined with compound glutamine enteric capsules has evident therapeutic effect on IBS-D of liver depression and spleen deficiency pattern, especially good at releasing abdominal distension and defecation urgency. The mechanism may be related with decreasing the levels of 5-HT and SP.