Effects of a Combination of Fu Zi and Rou Gui on Intestinal Neurotransmitters and Microflora in Rats with Slow Transit Constipation

[Objectives] This study was carried out to explore the combined effects of Fu Zi(Radix Aconiti Lateralis Praeparata, the secondary root of perennial herbaceous plant Acontium carmichaeli Dehx. of family Ranunculaceae) and Rou Gui(Cortex Cinnamomi, the bark of Cinnamamunz cassia Presl of family Lauraceae) on intestinal neurotransmitters and microflora in rats with slow transit constipation(STC). [Methods] Experimental rats were given loperamide hydrochloride by gavage to induce STC, and then treated with Fu Zi alone, Rou Gui alone, a combination of Fu Zi and Rou Gui(2:1 w/w), and prucalopride, respectively, for 14 days. Meanwhile, the general condition, the time to first black stool and the rate of intestinal propulsion of rats in each group were observed after STC was induced and after drug treatment, and the pathological changes in rat colon were observed via hematoxylin-eosin(HE) staining, and the levels of colonic 5-hydroxytryptamine(HT), vasoactive intestinal peptide(VIP) and substance P(SP) were detected by ELISA, and the changes in intestinal flora were detected by 16S rRNA Real-time PCR. [Results] Compared with healthy rats, the time to first black stool and the rate of intestinal propulsion, colonic 5-HT and SP levels significantly decreased(p<0.01), while their colonic VIP level significantly increased(p<0.01). Compared with STC rats, the time to first black stool, the rate of intestinal propulsion, colonic 5-HT and SP levels in Fu Zi-Rou Gui(2:1) treated rats and prucalopride treated rats significantly increased(p<0.01), while their colonic VIP level significantly decreased(p<0.01). There was no significant difference in alpha diversity between healthy rats and STC rats. However, analysis on beta diversity revealed that there were differences in microflora structure and composition between them. Compared with healthy rats, the relative abundance of Firmicutes and Proteobacteria in STC rats significantly increased, while that of Bacteroidetes decreased. Compared with STC rats, the relative abundance of Proteobacteria decreased and that of Bacteroidetes and Firmicutes increased in Fu Zi-Rou Gui(2:1) treated rats;the relative abundance of Bacteroidetes and Proteobacteria decreased while that of Firmicutes increased in Fu Zi treated rats;the relative abundance of Proteobacteria decreased while that of Bacteroidetes increased in Rou Gui treated rats;the relative abundance of Firmicutes and Proteobacteria decreased while that of Bacteroidetes increased in prucalopride treated rats. The intestinal flora in rats of all groups was dominated by Lactobacillus spp. and other genera of anaerobic bacteria. Compared with healthy rats, the relative abundance of Lactobacillus spp. and Clostridium spp. in STC rats decreased, while those of Blautia spp. and Ruminococcus spp. and Allobaculum spp. increased. Compared with STC rats, the relative abundance of Lactobacillus spp. in all rats treated with drugs increased. [Conclusions] The combination of Fu Zi and Rou Gui(2:1) can effectively improve intestinal motility in STC rats by regulating intestinal microbial community and the levels of colonic neurotransmitters.

Cinnamic acid regulates the intestinal microbiome and short-chain fatty acids to treat slow transit constipation

BACKGROUND Slow transit constipation(STC)is a disorder with delayed colonic transit.Cinnamic acid(CA)is an organic acid in natural plants,such as Radix Scrophulariae(Xuan Shen),with low toxicity and biological activities to modulate the intestinal microbiome.AIM To explore the potential effects of CA on the intestinal microbiome and the primary endogenous metabolites-short-chain fatty acids(SCFAs)and evaluate the therapeutic effects of CA in STC.METHODS Loperamide was applied to induce STC in mice.The treatment effects of CA on STC mice were assessed from the 24 h defecations,fecal moisture and intestinal transit rate.The enteric neurotransmitters:5-hydroxytryptamine(5-HT)and vasoactive intestinal peptide(VIP)were determined by the enzyme-linked immunosorbent assay.Hematoxylin-eosin and Alcian blue and Periodic acid Schiff staining were used to evaluate intestinal mucosa's histopathological performance and secretory function.16S rDNA was employed to analyze the composition and abundance of the intestinal microbiome.The SCFAs in stool samples were quantitatively detected by gas chromatography-mass spectrometry.RESULTS CA ameliorated the symptoms of STC and treated STC effectively.CA ameliorated the infiltration of neutrophils and lymphocytes,increased the number of goblet cells and acidic mucus secretion of the mucosa.In addition,CA significantly increased the concentration of 5-HT and reduced VIP.CA significantly improved the diversity and abundance of the beneficial microbiome.Furthermore,the production of SCFAs[including acetic acid(AA),butyric acid(BA),propionic acid(PA)and valeric acid(VA)]was significantly promoted by CA.The changed abundance of Firmicutes,Akkermansia,Lachnoclostridium,Monoglobus,UCG.005,Paenalcaligenes,Psychrobacter and Acinetobacter were involved in the production of AA,BA,PA and VA.CONCLUSION CA could treat STC effectively by ameliorating the composition and abundance of the intestinal microbiome to regulate the production of SCFAs.

A review on slow transit constipation

Slow transit constipation(STC),one of the most common chronic constipation in the clinic,is characterized by prolonged colonic transit time.Pathogenesis is unclear,and abnormalities can be found in enteric nervous system,intestinal neurotransmitters,intestinal smooth muscle,and Cajal stromal cell.Various methods have been used to improve the quality of life of patients with STC,including dietary fiber supplements,various cathartics,prokinetic drugs,and surgical intervention.However,there are still a large number of patients showing poor response to conservative treatment and intolerance to surgical treatment.Patients with STC suffer from severe constipation for long periods of time,faced with heavy economic burden and mental torture.Exploration the pathogenesis of STC to provide a clearer direction for future drug development is needed.

The efficacy and mechanism of fecal microflora transplantation in the treatment of slow transit constipation based on metabonomics anion mode

Background:Fecal microbiota transplantation(FMT)based on the positive ion mode of metabonomics has a good therapeutic benefit for slow transit constipation(STC)patients.However,a piece of comprehensive metabolomics information is yet to be established.The aim of the study was to explore the efficacy and mechanism of FMT in the treatment of STC under metabonomics.Methods:Eight STC patients meeting the set inclusion and exclusion criteria were enrolled and treated with FMT(three times).The Patient Assessment of Constipation-Symptoms(PAC-SYM),weekly total defecation times,and defecation frequency scores of these STC patients were compared before and after treatment.Feces and serum of STC patients before and after treatment were analyzed using 16SrDNA and metabolomics.Results:After FMT treatment,the PAC-SYM score of constipated patients decreased[(5.00±2.94)vs(5.20±2.87)],while the number of complete defecations per week increased[(2.00±1.79)vs(1.69±1.80)].The score of defecation frequency decreased[(0.83±1.03)vs(0.86±0.95)].The metabolites in the feces and serum of patients receiving FMT changed significantly(P<0.05).The results from 16SrDNA analysis showed that theαandβdiversity of the fecal microbiome changed significantly(P<0.05)after transplantation,and the contents of genera Lactobacillus,Bacillus,Succiniclasticum,Cellvibrio,and Escherichia increased in FMT treated patients.Conclusion:FMT may treat STC by increasing the beneficial intestinal flora and metabolites in the anion mode of metabolomics.

Role of Tenascin-X in regulating TGF-β/Smad signaling pathway in pathogenesis of slow transit constipation

BACKGROUND Chronic constipation is a gastrointestinal functional disease that seriously harms physical and mental health and impacts the quality of life of patients.Its incidence rate is 2%-27%.Slow transit constipation(STC)is a common type of chronic functional constipation,accounting for 10.3%-45.5%of such cases.Scholars have performed many studies on the pathogenesis of STC.These studies have indicated that the occurrence of STC may be related to multiple factors,such as dysfunction of the enteric nervous system,interstitial cells of Cajal(ICC)damage,and changes in neurotransmitters regulating intestinal peristalsis.AIM To investigate the role of Tenascin-X(TNX)in regulating the TGF-β/Smad signaling pathway in the pathogenesis of STC.METHODS This study included an experimental group and a control group.The experimental group included 28 patients with severe colonic STC,and the control group included 18 patients with normal colon tissues.Immunohistochemistry(IHC)was used to detect c-Kit,a specific marker of the ICC.Western blot,immunofluorescence,and IHC were used to detect the localization and expression of TNX and TGF-β/Smad.RESULTS IHC showed that the number of ICC with positive c-Kit expression was significantly reduced in the colon of STC patients(22.17±3.28 vs 28.69±3.53,P<0.05)and that the distribution was abnormal.Western blot results showed that c-Kit and Smad7 levels were significantly decreased in the colon of STC patients(c-kit:0.462±0.099 vs 0.783±0.178,P<0.01;Smad7:0.626±0.058 vs 0.799±0.03,P<0.01)and that TNX and Smad2/3 levels were higher in the STC group(TNX:0.868±0.028 vs 0.482±0.032,P<0.01).There was no significant difference in TGF-βbetween the two groups(0.476±0.028 vs 0.511±0.044,P=0.272).Pearson correlation analysis showed that the TNX protein exhibited a strong correlation with Smad2/3 and Smad7(P<0.05,|R|>0.8)and TGF-β(P<0.05,|R|=0.7).CONCLUSION The extracellular matrix protein TNX may activate the TGF-β/Smad signaling pathway by upregulating the Smad 2/3 signaling protein and thereby induce slight or complete epithelial stromal cell transformation,leading to an abnormal distribution and dysfunction of ICC in the diseased colon,which promotes the occurrence and development of STC.

Subtotal colonic bypass plus colostomy with antiperistaltic cecoproctostomy for the treatment of slow transit constipation in an aged population:A retrospective control study

AIM To compare the efficacy,improved quality of life,and prognosis in patients undergoing either subtotal colonic bypass with antiperistaltic cecoproctostomy(SCBAC) or subtotal colonic bypass plus colostomy with antiperistaltic cecoproctostomy(SCBCAC) for the treatment of slow transit constipation.METHODS Between October 2010 and October 2014,aged patients with slow transit constipation who were hospitalized and underwent laparoscopic surgery in our institute weredivided into two groups: the bypass group,15 patients underwent SCBAC,and the bypass plus colostomy group,14 patients underwent SCBCAC. The following preoperative and postoperative clinical data were collected: gender,age,body mass index,operative time,first flatus time,length of hospital stay,bowel movements(BMs),Wexner fecal incontinence scale,Wexner constipation scale(WCS),gastrointestinal quality of life index(GIQLI),numerical rating scale for pain intensity(NRS),abdominal bloating score(ABS),and ClavienDindo classification of surgical complications(CD) before surgery and at 3,6,12,and 24 mo after surgery.RESULTS All patients successfully underwent laparoscopic surgery without open surgery conversion or surgeryrelated death. The operative time and blood loss were significantly less in the bypass group than in the bypass plus colostomy group(P = 0.007). No significant differences were observed in first flatus time,length of hospital stay,or complications with CD > 1 between the two groups. No patients had fecal incontinence after surgery. At 3,6,and 12 mo after surgery,the number of BMs was significantly less in the bypass plus colostomy group than in the bypass group. The parameters at 3,6,12,and 24 mo after surgery in both groups significantly improved compared with the preoperative conditions(P < 0.05),except NRS at 3,6 mo after surgery in both groups,ABS at 12,24 mo after surgery and NRS at 12,24 mo after surgery in the bypass group. WCS,GIQLI,NRS,and ABS significantly improved in the bypass plus colostomy group compared with the bypass group at 3,6,12,and 24 mo after surgery(P < 0.05) except WCS,NRS at 3,6 mo after surgery and ABS at 3 mo after surgery. At 1 year after surgery,a barium enema examination showed that the emptying time was significantly better in the bypass plus colostomy group than in the bypass group(P = 0.007).CONCLUSION Laparoscopic SCBCAC is an effective and safe procedure for the treatment of slow transit constipation in an aged population and can significantly improve the prognosis. Its clinical efficacy is more favorable compared with that of SCBAC. Laparoscopic SCBCAC is a better procedure for the treatment of slow transit constipation in an aged population.

Slow transit constipation: A functional disorder becomes an enteric neuropathy

慢运输便秘传统地被看作了并且分类机能紊乱。然而，临床、压力计的证据一直在积累那建议大多数在 STC 的活动性改变怎么可能神经病的类型被考虑。另外，进一步的调查显示出那伤寒神经系统的微妙的改变，不是对常规组织学的检查明显，可能在这些病人在场。在现在的文章，我们将讨论这些证据，并且将试着与处于这个病理学的条件记录的大肠的反常运动机能把他们放在关系。

Normal aspects of colorectal motility and abnormalities in slow transit constipation

Human colonic motility is a relatively difficult topic to investigate. However, the refinement of manometric techniques in recent years enabled us to study both the proximal and distal segments of the viscus. The present paper reviews our knowledge about normal aspects of colorectal motility in man and the abnormalities found in slow transit constipation (STC), one of the most frequent and difficult to treat subtypes of constipation. An internetbased search strategy of the Medline and Science Citation Index was performed using the keywords colon, colonic,colorectal, constipation, slow transit, motility, recal, rectum in various combinations with the Boolean operators AND, OR and NOT. Only articles related to human studies were used, and manual cross-referencing was also performed.Most of colonic motor activity is represented by single nonpropagated contractions, rarely organized in bursts; this activity is maximal during the day, especially after waking and following meals. In addition, a specialized propagated activity with propulsive features is detectable, represented by high- and low-amplitude propagated contractions. In the severe form of constipation represented by the slow transit type, the above motor activity is completely deranged. In fact, both basal segmental activity (especially in response to meals) and propagated activity (especially that of high amplitude) are usually decreased, and this may represent a physiologic marker of this disorder. Human colonic motor activity is quite a complex issue, still only partly understood and investigated, due to anatomic and physiological difficulties. In recent years, however, some more data have been obtained, even in proximal segments. These data have helped in elucidating, althoughonly in part, some pathophysiological mechanisms of chronic constipation, and especially of the STC subtype.

Preservation of superior rectal artery in laparoscopically assisted subtotal colectomy with ileorectal anastomosis for slow transit constipation

BACKGROUND Slow transit constipation(STC)has traditionally been considered as a functional disorder.However,evidence is accumulating that suggests that most of the motility alterations in STC might be of a neuropathic etiology.If the patient does not meet the diagnosis of pelvic outlet obstruction and poorly response to conservative treatment,surgical intervention with subtotal colectomy may be effective.The most unwanted complication of the procedure is anastomotic leakage,however,preservation of the superior rectal artery(SRA)may reduce its incidence.AIM To evaluate the preservation of the SRA in laparoscopically assisted subtotal colectomy with ileorectal anastomosis in STC patients.METHODS This was a single-center retrospective observational study.STC was diagnosed after a series of examinations which included a colonic transit test,anal manometry,a balloon expulsion test,and a barium enema.Eligible patients underwent laparoscopically assisted total colectomy with ileorectal anastomosis and were examined between January 2016 and January 2018.The operation time,blood loss,time to first flatus,length of hospital days,and incidence of minor or major complications were recorded.RESULTS A total of 32 patients(mean age,42.6 years)who had received laparoscopic assisted subtotal colectomy with ileorectal artery anastomosis and preservation of the SRA.All patients were diagnosed with STC after a series of examinations.The mean operative time was 151 min and the mean blood loss was 119 mL.The mean day of first time to flatus was 3.0 d,and the mean hospital stay was 10.6 d.There were no any patients conversions to laparotomy.Post-operative minor complications including 1 wound infection and 1 case of ileus.There was no surgical mortality.No anastomosis leakage was noted in any of the patients.CONCLUSION Laparoscopically assisted subtotal colectomy with ileorectal anastomosis and preservation of the SRA can significantly improve bowel function with careful patient selection.Sparing the SRA may protect against anastomosis leakage.

STUDY ON GASTROINTESTINAL MOTILITY IN SLOW TRANSIT CONSTIPATION

Objective To investigate motor activity of gastrointestinal tract in patients with slow transit constipation. Methods 42 patients with STC and 20 healthy controls were included in the study. Each subject underwent colonic transit test, gastric emptying, orocecal transit time, electromyography and anorectal manometry. Results According to transit index, 42 STC patients were divided into 3 types: ①0.5<TI≤1.0, slow rectosigmoid transit 15 cases; ②TI=0.5, colorectal stasis 10 cases; ③0<TI≤0.5, slow colonic transit 17 cases. Gastric emptying rate at the fourth hour was decreased in STC patients than in controls . The mean orocaecal transit time was significantly delayed in the patients . Paradoxical anal sphincter contraction with defecation effort was higher prevalent in patients with slow rectosigmoid transit and colorectal stasis than patients with slow colonic transit. Minimum relaxation volume and maximal rectal tolerable volume were significantly increased in STC patients . Conclusion STC displayed colonic abnormality, and/or pelvic floor dysfunction. In addition, it showed motor abnormalities of upper gut, might be part of a pan enteric motor disorders.

CLINICAL OBSERVATION ON THE TREATMENT OF 34 CASES OF SLOW TRANSIT CONSTIPATION WITH ELECTROACUPUNCTURE

Objective: To explore effective modality for slow transit constipation (STC) and provide evidence for standardization of pattern identification of STC. Methods: Sixty eight cases of STC patients were randomly and evenly assigned into acupuncture group (n=34) and medication (control) group (n=34). In acupuncture group, Tianshu (天枢ST 25) and Zhigou (支沟TE 6) were punctured and stimulated electrically by setting the parameters of 1 Hz, continuous waves, suitable strength and duration of 30 min. Patients of the control group were ordered to take Tongbianling (bowel-movement promotion) Capsule, 5 capsules/time, once daily. Results: After 28 days’ treatment, symptom scores in both groups were decreased significantly and colonic ejection rates within 72 hours increased considerably in comparison with pre-treatment (P<0.01); and the therapeutic effects of acupuncture group were remarkably better than those of control group in lowering symptom score and raising colonic ejection rate (P<0.01). Conclusion: Acupuncture is a simple and practical method for the treatment of STC and its therapeutic effect is reliable.

Colectomy for idiopathic slow transit constipation

Objective: To evaluate the intervention of colectomy on a group of patients with idiopathic slow transit constipation (STC).Methods: Thirty-four patients with STC, underwent colectomy during recent 10 years in our department, were subjected and followed for a mean length of 34 months, and their colon transits, defecograms, colonoscopic examination, sex hormone detection, and immunohistochemical studies were retrospectively reviewed.Results: The colonic transit time ranged from 96 to 240 h, with a mean time of 136 h.Eighty-five percent of patients (29/34) accompanied with outlet obstructed constipation, and 50% (17/34) showed abnormal sex hormone levels.Colectomy obtained satisfactory results in most patients, except one case of recurrence.Moreover, more neurons positive to nitric oxide synthase (NOS) and lesser to vasoactive intestinal polypeptide (VIP) were seen in the colonic myenteric plexus.Conclusion: Colectomy produces a satisfactory functional outcome in the majority of patients undergoing surgery for slow transit constipation, but accompanied pelvic dysfunction must be corrected simultaneously.

EVALUATION ON THE THERAPEUTIC EFFECT OF DEEP PUNCTURE OF TIANSHU (ST 25) FOR SLOW TRANSIT CONSTIPATION

Objective: To evaluate the short-term and middle-long-term therapeutic effects of deep puncture of Tianshu (天枢ST 25) for slow transit constipation (STC). Methods: Sixty cases of STC patients were evenly randomized into acupuncture group and medication group. In acupuncture group, electroacupuncture (EA, 20 Hz, continuous waves and tolerable strength) was applied to bilateral Tianshu (ST 25) for 30 min after deep puncture. The treatment was given once daily, 5 sessions every week, two weeks altogether. Patients of control group were ordered to take Lactulose (10 mL/time, b.i.d), two weeks altogether. Cleveland Constipation Score (CCS) and colonic transit time (CCT) were used to evaluate the therapeutic effect. Results: After treatment, both CCS and CCT showed that the therapeutic effect of acupuncture group was significantly superior to that of medication group (P<0.05). Six-months’ follow up showed that the therapeutic effect of acupuncture still maintained in 13 of the 22 cases visited, while that of control group only maintained in 1 of the 19 cases. Conclusion: The therapeutic effect of deep puncture of Tianshu (ST 25) for STC is definite and has a middle-long-term effect. The patients have no any unfavorable reactions.

Proteomic analysis of down-regulated proteins in colonic mucosa of chronic slow transit constipation rats

Objective:To investigate the alternations of proteins in the colonic mucosa of chronic slow transit constipation(STC) rats with a 2-DE-based proteomic method and analyze the function of these down-regulated proteins so as to provide theoretical basis for the pathogenesis of intestinal mucosa of chronic STC rats.Methods:STC model was established by feeding rats with 8 mg/(kg·d) diphenoxylate for 120 d.An experimental model of chronic STC rat was used for separation of proteomics from colonic mucosa using two-dimensional electrophoresis(2-DE).Proteins altered in expressional level were identified by Image Master 2DElite,mass spectrometry,and bibliometrics were applied to identify the differential protein expression and their clinical significance and function were analyzed.Results:Obvious differential protein expression was observed in the pathogenesis of STC,including mast cell protease(A1),non-specific dipeptidase(A2) and chondrosome succinate dehydrogenase precursor(A3).The expressions of A1,A2 and A3 were down-regulated in the gel graph of STC rats.Conclusion:The down-regulation of chondrosome succinate dehydrogenase,mast cell protease as well as non-specific dipeptidase in rat colon suggests the functional impairment of the oxidoreduction of mitochondrion is very important in the genesis and development of STC.The immunological reaction of STC rats is weakened,and the function of digesting and absorbing protein may be damaged to some extent.

健脾理气润肠法治疗STC对患者临床症状及肠神经递质水平的影响

目的研究健脾理气润肠法应用于慢传输型便秘(STC)患者中的临床效果。方法选取2021年8月至2022年12月上海市静安区中医医院肛肠科收治的120例STC患者作为研究对象,采用随机数表法将所有患者分为两组各60例,跟踪随访1个月后,脱落5例,最终115例患者完成研究。其中58例接受乳果糖口服液治疗者设为对照组,57例接受健脾理气润肠汤联合乳果糖口服液治疗者设为观察组。比较两组患者治疗1个月后的治疗效果,以及入院时、治疗7 d、治疗1个月后的便秘主要性状评分、便秘患者症状自评量表(PAC-SYM)评分、Bristol评分、肠神经递质水平[生长抑素(SS)、血管活性肠肽(VIP)、P物质(SP)]和治疗24 h、48 h、72 h的结肠传输试验标志物残留率。结果观察组患者的治疗总有效率为98.25%,明显高于对照组的86.21%,差异有统计学意义(P<0.05);两组患者入院时的便秘主要症状评分比较差异均无统计学意义(P>0.05);治疗7 d、1个月后,观察组患者的各项便秘主要症状评分明显低于对照组,差异均有统计学意义(P<0.05);治疗7 d和1个月后,观察组患者的Bristol评分分别为(21.35±5.69)分、(15.63±3.88)分,明显低于对照组的(28.96±4.19)分、(20.20±4.52)分,PAC-SYM评分分别为(5.96±0.88)分、(6.33±0.76)分,明显高于对照组的(4.55±0.87)分、(5.69±0.74)分,差异均有统计学意义(P<0.05);治疗7 d、1个月后,观察组患者的SS水平分别为(18.52±3.44)ng/mL、(14.20±3.52)ng/mL,明显低于对照组的(21.20±3.69)ng/mL、(18.41±3.66)ng/mL,VIP水平分别为(36.52±5.58)ng/mL、(25.63±7.41)ng/mL,明显低于对照组的(40.10±5.20)ng/mL、(31.20±6.85)ng/mL,SP水平分别为(40.52±6.88)ng/mL、(55.52±7.41)ng/mL,明显高于对照组的(35.63±4.20)ng/mL、(44.52±8.52)ng/mL,差异均有统计学意义(P<0.05);治疗24 h、48 h、72 h后观察组患者的肠传输实验标志物残留率分别为(60.00±4.20)%、(30.00±5.10)%、(15.00±2.21)%,明显低于对照组的(68.75±5.52)%、(40.00±5.55)%、(20.00±2.55)%,差异均有统计学意义(P<0.05)。结论健脾理气润肠法应用于STC患者中,可明显改善肠神经递质水平,患者短时间内临床症状得以缓解,且传输实验标志物残留率低,利于减轻不适感。

Consensus statement AIGO/SICCR:Diagnosis and treatment of chronic constipation and obstructed defecation(partⅠ:Diagnosis)

Chronic constipation is a common and extremely troublesome disorder that significantly reduces the quality of life,and this fact is consistent with the high rate at which health care is sought for this condition.The aim of this project was to develop a consensus for the diagnosis and treatment of chronic constipation and obstructed defecation.The commission presents its results in a "Question-Answer" format,including a set of graded recommendations based on a systematic review of the literature and evidence-based medicine.This section represents the consensus for the diagnosis.The history includes information relating to the onset and duration of symptoms and may reveal secondary causes of constipation.The presence of alarm symptoms and risk factors requires investigation.The physical examination should assess the presence of lesions in the anal and perianal region.The evidence does not support the routine use of blood testing and colonoscopy or barium enema for constipation.Various scoring systems are available to quantify the severity of constipation;the Constipation Severity Instrument for constipation and the obstructed defecation syndrome score for obstructed defecation are the most reliable.The Constipation-Related Quality of Life is an excellent tool for evaluating the patient's quality of life.No single test provides a pathophysiological basis for constipation.Colonic transit and anorectal manometry define the pathophysiologic subtypes.Balloon expulsion is a simple screening test for defecatory disorders,but it does not define the mechanisms.Defecography detects structural abnormalities and assesses functional parameters.Magnetic resonance imaging and/or pelvic floor sonography can further complement defecography by providing information on the movement of the pelvic floor and the organs that it supports.All these investigations are indicated to differentiate between slow transit constipation and obstructed defecation because the treatments differ between these conditions.

Zhizhu Decoction Alleviates Intestinal Barrier Damage via Regulating SIRT1/FoxO1 Signaling Pathway in Slow Transit Constipation Model Mice

Objective:To explore the possible effects and mechanism of Zhizhu Decoction(ZZD)on the pathophysiology of slow transit constipation(STC).Methods A total of 54 C57BL/6 mice was randomly divided into the following 6 groups by a random number table,including control,STC model(model),positive control,and low-,medium-and high-doses ZZD treatment groups(5,10,20 g/kg,namely L,M-,and H-ZZD,respectively),9 mice in each group.Following 2-week treatment,intestinal transport rate(ITR)and fecal water content were determined,and blood and colon tissue samples were collected.Hematoxylin-eosin and periodic acid-Schiff staining were performed to evaluate the morphology of colon tissues and calculate the number of goblet cells.To determine intestinal permeability,serum levels of lipopolysaccharide(LPS),low-density lipoprotein(LDL)and mannose were measured using enzyme-linked immunosorbent assay(ELISA).Western blot analysis was carried out to detect the expression levels of intestinal tight junction proteins zona-occludens-1(ZO-1),claudin-1,occludin and recombinant mucin 2(MUC2).The mRNA expression levels of inflammatory cytokines including tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,IL-4,IL-10 and IL-22 were determined using reverse transcription-quantitative reverse transcription reaction.Colon indexes of oxidative stress were measured by ELISA,and protein expression levels of colon silent information regulator 1/forkhead box O transcription factor 1(SIRT1/FoxO1)antioxidant signaling pathway were detected by Western blot.Results Compared with the model group,ITR and fecal moisture were significantly enhanced in STC mice in the M-ZZD and H-ZZD groups(P<0.01).Additionally,ZZD treatment notably increased the thickness of mucosal and muscular tissue,elevated the number of goblet cells in the colon of STC mice,reduced the secretion levels of LPS,LDL and mannose,and upregulated ZO-1,claudin-1,occludin and MUC2 expressions in the colon in a dose-dependent manner,compared with the model group(P<0.05 or P<0.01).In addition,ZZD significantly attenuated intestinal inflammation and oxidative stress and activated the SIRT1/FoxO1 signaling pathway(P<0.05 or P<0.01).Conclusion ZZD exhibited beneficial effects on the intestinal system of STC mice and alleviated intestinal inflammation and oxidative stress via activating SIRT1/FoxO1 antioxidant signaling pathway in the colon.

不同频率电针介导PINK1/Parkin通路治疗STC模型大鼠的机制研究

目的 探讨不同频率电针介导线粒体自噬相关蛋白(PINK1)/帕金蛋白(Parkin)通路治疗慢传输型便秘(STC)模型大鼠的作用。方法 取34只8周龄雄性健康SD大鼠,采用随机数字表分为正常对照组7只和建模组27只。建模组采用洛哌丁胺混悬液灌胃法制备STC模型大鼠,建模成功后采用随机数字表分为模型对照组,2 Hz、50 Hz电针组和假电针组。2 Hz、50 Hz电针组予以电子针疗仪频率2 Hz、50 Hz,强度10 mA刺激“后三里”,假电针组针刺“后三里”但不通电,模型对照组和正常对照组仅固定处理,每天30 min,每天1次,持续14 d。碳素墨汁灌胃法检测肠道推进率;苏木素-伊红(HE)染色观察结肠组织病理改变;免疫荧光共聚焦显微镜观察结肠Cajal间质细胞(ICC)形态;免疫组化及蛋白质免疫印迹(WB)法检测结肠组织原癌基因(C-Kit)、PINK1、Parkin、自噬效应蛋白1(Beclin1)、核孔糖蛋白(p62)、自噬相关蛋白3(LC3)蛋白表达。结果 与模型对照组、假电针组比较,50 Hz电针组、2 Hz电针组增加肠道推进率[(60.63±10.21)%、(55.84±9.04)%比(48.43±7.02)%、(49.60±8.19)%,P<0.05]、ICC数量[(29.78±4.19)个、(20.45±4.37)个比(10.50±2.11)个、(11.02±2.28)个,P<0.05]、免疫荧光积分光密度(IOD)值[(2101.35±320.33)、(1936.04±307.26)比(1615.82±276.33)、(1635.97±281.52),P<0.05]、C-kit蛋白表达[(1.26±0.21)、(0.92±0.17)比(0.45±0.07)、(0.48±0.09),P<0.05]、p62蛋白表达[(1.06±0.19)、(0.87±0.13)比(0.54±0.09)、(0.55±0.10),P<0.05]。与模型对照组、假电针组比较,50 Hz电针组、2 Hz电针组降低PINK1蛋白表达[(0.37±0.08)、(0.59±0.11)比(0.85±0.17)、(0.83±0.15),P<0.05]、Parkin蛋白表达[(0.55±0.09)、(0.78±0.12)比(1.02±0.21)、(1.04±0.22),P<0.05]、Beclin1蛋白表达[(0.61±0.11)、(0.90±0.13)比(1.07±0.21)、(1.09±0.23),P<0.05]、LC3蛋白表达[(0.69±0.12)、(0.82±0.15)比(1.14±0.18)、(1.11±0.20),P<0.05]。正常对照组大鼠结肠组织、ICC形态均正常,模型对照组和假电针组均呈显著改变,2 Hz电针组和50 Hz电针组均改善,且50 Hz电针组基本接近正常对照组。结论 电针“后三里”能够减轻STC大鼠结肠模型病理改变和ICC形态学改变,且可能可下调PINK1、Parkin、Beclin1、LC3表达,上调Ckit、p62表达,其中50 Hz电针作用更佳。

电针对慢传输型便秘大鼠肠道运动及促炎性因子表达影响

目的观察电针下合穴“足三里”“上巨虚”对慢传输型便秘(STC)大鼠肠道运动及促炎性因子表达的影响。方法24只健康清洁级SD大鼠随机分为空白组、模型组、药物组和电针组,每组各6只。采用复方地芬诺酯混悬液10 mg·kg^(-1)剂量灌胃制备STC大鼠模型,每天1次,连续14 d。空白组予等剂量0.9%氯化钠溶液灌胃。电针组给予双侧“足三里”“上巨虚”电针治疗,选择疏密波,频率2/50 Hz,电流大小以局部肌肉轻微抖动为度,每次治疗20 min,每天1次,连续治疗14 d。药物组予枸橼酸莫沙必利片混悬液按照1.37 mg·kg^(-1)、1 mL·100 g^(-1)剂量灌胃治疗,疗程同上。观察各组治疗后首粒黑便排出时间、小肠推进率,采用HE染色观察结肠组织病理形态学变化,采用Western-blot法检测肠道肿瘤坏死因子α(TNF-α)、白介素1β(IL-1β)、CD68蛋白表达。结果模型组大鼠首粒黑便排出时间较空白组显著加长,小肠推进率显著降低(P<0.05);药物组和电针组大鼠经治疗后,与模型组比较,首粒黑便排出时间显著减少、小肠推进率显著提高(P<0.05)。HE染色可见造模后大鼠结肠黏膜组织出血,组织形态学结构破坏,伴上皮细胞变性坏死及炎性细胞浸润;电针和药物治疗能够减少炎性细胞浸润,改善组织形态学变化。Western-blot结果显示,模型组大鼠TNF-α、IL-1β、CD68蛋白表达较空白组显著升高(P<0.05);药物组和电针组大鼠经治疗后,结肠组织TNF-α、IL-1β、CD68蛋白表达较模型组显著降低(P<0.05)。结论电针治疗能够改善STC大鼠肠道运动功能,修复结肠组织病理形态学改变,其机制可能与下调TNF-α、IL-1β、CD68蛋白表达,抑制炎性反应有关。

生血通便颗粒对血虚肠燥型慢传输型便秘大鼠结肠肌电及Ca^(2+)/CaM/MLCK信号通路的影响

目的观察生血通便颗粒对血虚肠燥型慢传输型便秘(STC)大鼠结肠肌电及Ca^(2+)/CaM/MLCK信号通路的影响,探讨其治疗STC的作用机制。方法采用洛哌丁胺灌胃结合尾部放血法建立血虚肠燥型STC大鼠模型。将大鼠分为对照组、模型组、枸橼酸莫沙必利组和生血通便颗粒组,每组8只,给药组分别予相应药物灌胃,连续14 d。观察大鼠治疗前后一般状况,检测大鼠粪便含水量,生物机能实验系统检测结肠肌电慢波频率、振幅及变异系数,测定肠道推进率,HE染色观察结肠组织病理变化,比色法检测结肠平滑肌细胞Ca^(2+)浓度,Western blot检测结肠平滑肌组织缝隙连接蛋白43(Cx43)、钙调蛋白(CaM)、肌球蛋白轻链激酶(MLCK)、磷酸化肌球蛋白轻链20(p-MLC20)蛋白表达。结果与对照组比较,模型组大鼠体质量、粪便含水量和肠道推进率显著降低(P<0.01),结肠肌电慢波频率减慢、频率变异系数增加(P<0.01),慢波振幅和振幅变异系数增加(P<0.01);结肠黏膜结构受损,可见炎性改变,糜烂明显,结肠平滑肌细胞内Ca^(2+)浓度及Cx43、CaM、MLCK、p-MLC20蛋白表达显著降低(P<0.01)。与模型组比较,枸橼酸莫沙必利组和生血通便颗粒组大鼠体质量、粪便含水量和肠道推进率显著升高(P<0.05,P<0.01),结肠肌电慢波频率加快、频率变异系数减小(P<0.01),慢波振幅和振幅变异系数减小(P<0.05,P<0.01);结肠黏膜结构较完整,糜烂情况改善,结肠平滑肌细胞内Ca^(2+)浓度及Cx43、CaM、MLCK、p-MLC20蛋白表达显著升高(P<0.01)。结论生血通便颗粒可改善血虚肠燥型STC大鼠症状,恢复结肠动力,其机制可能与调节结肠肌电节律性、激活Ca^(2+)/CaM/MLCK信号通路相关。