A study of acupoint specificity and mechanism of electroacupuncture intervention on chronic colitis in rats based on PI3K/AKT/mTOR signaling pathway

Objective:This study aimed to elucidate the differences in effects and mechanisms of action of electric-needle therapy at Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)and Shang Ju Xu(ST37)acupoints on chronic experimental colitis in rats through the PI3K/AKT/mTOR signaling pathway.Methods:Sixty pathogen-free SD rats were randomly assigned to six groups:the normal,model,Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)and Shang Ju Xu(ST37)groups,each with 10 rats.Chronic colitis was induced in rats by combining immunization and local stimulation.After model establishment,electrical needle intervention combined with dispersing wave of 2 Hz/50 Hz with a current intensity of 2 mA once daily for 20 min was applied on acupoints of each group.Subsequently,the inflammation of colonic mucosa and serum levels of inflammatory factors(IL-23,IL-17,IL-10)were observed;ELISA was used to detect mRNA expressions of PI3K,Akt and mTOR in colitic tissues by RT-PCR as well as protein content of p-PI3k/PI3K,p-Akt/Akt,and p-mTOR/mTOR in colitic tissues by Western blotting.Result:Compared with the normal group,the model rats showed a poor general condition,serious damage to the colonic mucosa with a large number of inflammatory cells infiltration.The serum IL-23 and IL-17 expressions were significantly increased(P<0.01),while the serum IL-10 expression was significantly decreased(P<0.01);the mRNA and protein expressions of PI3K,Akt,mTOR and p-PI3K,p-Akt and p-mTOR were significantly increased(P<0.05,P<0.01).Compared with the model group,the pathological slices of rats in each acupoints intervention group showed obvious improvement of colitis inflammatory reaction and tissue damage;the serological levels of IL-23 and IL-17 were significantly reduced(P<0.01),while the serology level of IL-10 was significantly increased(P<0.01);the expressions of PI3K,Akt,mTOR mRNA and p-PI3K,p-Akt,p-mTOR proteins were significantly decreased(P<0.05,P<0.01).Compared with Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)groups,the recovery degree of mucosa layers in Shang Ju Xu(ST37)group was closer to that of normal group,and the curative effect was relatively the best;in terms of serological levels of IL-23 and IL-17,the Shang Ju Xu(ST37)group was significantly lower(P<0.05),while the level of IL-10 was significantly higher(P<0.01);the expressions of PI3K,Akt,mTOR mRNA and p-PI3K,p-Akt,p-mTOR proteins were significantly decreased(P<0.05,P<0.01).Conclusion:Results indicate that electrical acupuncture at Tian Shu(ST25),Da Chang Shu(BL25),Zu San Li(ST36)and Shang Ju Xu(ST37)show similar effects in relieving the colitis-induced damage in the mucosa of chronic colitis rats,as well as inflammatory response.Among them,Shang Ju Xu(ST25)has a superior overall effect in treating chronic colitis compared to Tian Shu(ST25),Da Chang Shu(BL25)and Zu San Li(ST36).The mechanism may be related to inhibition of PI3K/Akt/mTOR signaling pathway.

Sodium selenite ameliorates dextran sulfate sodiuminduced chronic colitis in mice by decreasing Th1, Th17, and γδT and increasing CD4(+)CD25(+) regulatory T-cell responses

AIM To assess the effect of sodium selenite on the severity of dextran sulfate sodium(DSS)-induced colitis in C57BL/6 mice.METHODS Mice were randomly divided into four groups(n = 10/group): normal group, selenium(Se) group, chronic colitis group, and Se + chronic colitis group. The mice were sacrificed on day 26. Survival rates, clinical symptoms, colon length, and histological changes were determined. The percentages and absolute numbers of immune system cells in the lamina propria lymphocytes(LPL) of the colon, the expression of m RNA in colon tissue, and the concentrations of Th1, Th17, and Treg cytokines in LPL from the large intestine, were measured.RESULTS Se significantly ameliorated the symptoms of colitis and histological injury(P < 0.05 each), increasing the proportions of neutrophils and CD4+ CD25+ T cells(P < 0.05 each) and decreasing the proportions of γδT cells, CD4+, CD4+CD44+, and CD4+ CD69+ T cells in LPL(P < 0.05 each). Moreover, Se reduced the expression of IL-6, IFN-γ, IL-17 A, IL-21, T-bet, and RORγt(P < 0.05 each), but enhanced the expression of IL-10 and Foxp3(P < 0.05 each). CONCLUSION These results suggest that Se protects against DSSinduced chronic colitis perhaps by increasing the number of CD4(+)CD25(+) Tregs that suppress the secretion of proinflammatory cytokines and populations of Th1, Th17, and γδT cells.

Infliximab does not increase colonic cancer risk associated to murine chronic colitis

AIM To explore the influence of Infliximab(IFX) on cancer progression in a murine model of colonic cancer associated to chronic colitis.METHODS AOM/DSS model was induced in C57BL/6 mice. Mice were injected with IFX(5 mg/kg) during each DSS cycle while control mice received saline. Body weight, occult blood test and stool consistency were measured to calculate the disease activity index(DAI). Mice were sacrificed at week 10 and colons were analyzed macroscopically and microscopically for number of cancers and degree of inflammation. MTT assay was performed on CT26 to evaluate the potential IFX role on metabolic activity and proliferation. Cells were incubated with TNF-α or IFX or TNF-α plus IFX, and cell vitality was evaluated after 6, 24 and 48 h. The same setting was used after pre-incubation with TNF-α for 24 h.RESULTS IFX significantly reduced DAI and body weight loss in mice compared with controls, preserving also colon length at sacrifice. Histological score was also reduced in treated mice. At macroscopic analysis, IFX treated mice showed a lower number of tumor lesions compared to controls. This was confirmed at microscopic analysis, although differences were not statistically significant. In vitro, IFX treated CT26 maintained similar proliferation ability at MTT test, both when exposed to IFX alone and when associated to TNF-α.CONCLUSION IFX did not increase colonic cancer risk in AOM-DSS model of cancer on chronic colitis nor influence directly the proliferation of murine colon cancer epithelial cells.

Clinical Efficacy on the Treatment of Chronic Colitis with Feng-Liao-Chang-Wei-Kang Granules Combined with Mesalazine

[Objectives]To analyze the clinical effect of Feng-Liao-Chang-Wei-Kang Granule combined with mesalazine on chronic colitis.[Methods]120 patients with chronic enteritis admitted to Qionghai Hospital of Traditional Chinese Medicine from October 2020 to March 2022 were selected as the research objects,and were randomly divided into combined group(62 cases)and conventional group(58 cases).Patients in the conventional group were treated with oral mesalazine on the basis of conventional treatment,while patients in the combined group were treated with Feng-Liao-Chang-Wei-Kang Granule combined with mesalazine.The clinical efficacy,inflammatory factor level,colonic mucosal lesion degree,quality of life and TCM syndrome scores of the two groups were compared.[Results]The total clinical effective rate in the combined group was higher than that in the conventional group(P<0.05).After treatment,the levels of inflammatory factors in the two groups were lower than those before treatment,and the levels of interleukin-6(IL-6),C-reactive protein(CRP)and tumor necrosis factor-α(TNF-α)in the combined group were lower than those in the conventional group(P<0.001).After treatment,the Baron and Geboes scores of the two groups were lower than those before treatment,and the Baron and Geboes scores of the combined group were lower than those of the conventional group(P<0.001).After treatment,the scores of quality of life in the two groups were higher than those before treatment,while the scores of TCM syndromes were lower than those before treatment.The scores of quality of life in the combined group were higher than those in the conventional group(P<0.001).[Conclusions]Feng-Liao-Chang-Wei-Kang Granule combined with mesalazine can obviously improve the curative effect,reduce the level of inflammatory factors,improve the degree of colonic mucosal lesions and improve the quality of life of patients with chronic colitis,which is worthy of popularization and application in clinical practice.

IN SITU IMMUNOHISTOCHEMISTRY STUDY OF INTERCELLULAR ADHESION MOLECULE－1（ICAM－1） IN CHRONIC COLITIS AND COLON CANCER

Cell adhesion moleculaes(CAMs) are related to many pathologic processes.In this paper, in situ .expression of intereellular adhesion molecule-1 (ICAM-1,CD54) in chronic colitis and colon cancer- psmpared and investlgatfd using lmmunohistochemical method. The result showed that,tompared with normal colon mucosa,a lot of ICAM-1+ cells were seen in the mucosa of chronic colitis, but muck fewer ICAM-1+ cells were found in colon cancer tissue.Also,the degree of ICAM-1+ cell decrease was consistent with that or cancer cell differentiation. All the findings suggest that ICAM-1 may be related to the local immunological tolerance of colon cancer.

Efficacy and safety of stellate ganglion block in chronic ulcerative colitis

Sympathetic system modulation by stellate ganglion blockade may modulate immune dysfunction and significantly improve symptoms of chronic ulcerative colitis.

From the imbalance of tumor amino acid metabolism,the application of fuzheng detoxification method in interfering with the transformation of colitis cancer was discussed

Objective:From the perspective of tumor amino acid metabolism imbalance,the application of fuzheng detoxification method in intervening in the transformation of colitis cancer was discussed.Methods:By consulting CNKI's concept of tumor microenvironment,energy metabolism,amino acid metabolism,and colitis cancer transformation,it provides theoretical and data support for research purposes.Results:The study found that the transformation of colitis cancer refers to the metabolic imbalance of the tumor microenvironment under the condition of repeated stimulation of chronic inflammatory responses of colon tissue,which is mainly related to energy metabolism,sugar metabolism,fat metabolism,and amino acid metabolism imbalance,which promotes the transformation of the inflammatory microenvironment to the tumor microenvironment.Amino acids as an important source of nutrients for cells,its metabolic imbalance is an important cause of inflammation and cancer transformation.Conclusion:Chinese medicine believes that the transformation of inflammation and cancer is mainly related to the incurable treatment of evil in the internal organs,the loss of Qi,and the poison damage to the blood network.Colon cancer disease mechanism is always positive deficiency and stasis,the pathogenesis is based on the virtual standard,the virtual and real are mixed,and the clinical use of correct detoxification to adjust the balance of the body.

In-vivo characterization of DALM in ulcerative colitis with high-resolution probe-based confocal laser endomicroscopy

Recently,the use of confocal laser endomicroscopy(CLE) in the diagnosis of chronic ulcerative colitis(CUC) was reported.In this brief report we aimed to assess the application of probe-based CLE to characterize colonic mucosa and dysplasia in CUC.The study involved a patient presenting long-standing CUC.Confocal imaging of both the inflamed mucosa,a circumscribed lesion(dysplasiaassociated lesional mass),and adjacent colonic mucosa are demonstrated and the correlation between the CLE and histological images.Inflamed mucosa and dysplasia showed specific alteration of crypt architecture,cellular infiltration,and vessel architecture with an excellent correlation between CLE and standard histological examination.

Juvenile ferric iron prevents microbiota dysbiosis and colitis in adult rodents

AIM: To assess whether juvenile chronic ferric iron ingestion limit colitis and dysbiosis at adulthood in rats and mice. METHODS: Two sets of experiments were designed. In the first set, recently weaned mice were either orally administered ferrous (Fe2+) iron salt or ferric (Fe3+) microencapsulated iron for 6 wk. The last week of experiments trinitrobenzene sulfonic acid (TNBS) colitis was induced. In the second set, juvenile rats received the microencapsulated ferric iron for 6 wk and were also submitted to TNBS colitis during the last week of experiments. In both sets of experiments, animals were sacrificed 7 d after TNBS instillation. Severity of the inflammation was assessed by scoring macroscopic lesions and quantifying colonic myeloperoxidase (MPO) activity. Alteration of the microflora profile was estimated usingquantitative polymerase chain reaction (qPCR) by measuring the evolution of total caecal microflora, Bacteroidetes, Firmicutes and enterobacteria. RESULTS: Neither ferrous nor ferric iron daily exposures at the juvenile period result in any effect in control animals at adulthood although ferrous iron repeated administration in infancy limited weight gain. Ferrous iron was unable to limit the experimental colitis (1.71 ± 0.27 MPO U/mg proteinvs 2.47 ± 0.22 MPO U/mg protein in colitic mice). In contrast, ferric iron significantly prevented the increase of MPO activity (1.64 ± 0.14 MPO U/mg protein) in TNBS-induced colitis. Moreover, this positive effect was observed at both the doses of ferric iron used (75 and 150 mg/kg per day po - 6 wk). In the study we also compared, in both rats and mice, the consequences of chronic repeated low level exposure to ferric iron (75 mg/kg per day po - 6 wk) on TNBS-induced colitis and its related dysbiosis. We confirmed that ferric iron limited the TNBS-induced increase of MPO activity in both the rodent species. Furthermore, we assessed the ferric iron incidence on TNBS-induced intestinal microbiota dysbiosis. At first, we needed to optimize the isolation and quantify DNA copy numbers using standard curves to perform by qPCR this interspecies comparison. Using this approach, we determined that total microflora was similar in control rats and mice and was mainly composed of Firmicutes and Bacteroidetes at a ratio of 10/1. Ferric juvenile administration did not modify the microflora profile in control animals. Total microflora numbers remained unchanged whichever experimental conditions studied. Following TNBS-induced colitis, the Firmicutes/Bacteroidetes ratio was altered resulting in a decrease of the Firmicutes numbers and an increase of the Bacteroidetes numbers typical of a gut inflammatory reaction. In parallel, the subdominant population, the enterobacteria was also increased. However, ferric iron supplementation for the juvenile period prevented the increase of Bacteroidetes and of enterobacteria numbers consecutive to the colitis in both the studied species at adulthood.CONCLUSION: Rats and mice juvenile chronic ferric iron ingestion prevents colitis and dysbiosis at adulthood as assessed by the first interspecies comparison.

健脾理肠片免疫作用的临床与实验研究

用健脾理肠片治疗慢性结肠炎及溃疡性结肠炎50例,并设对照组(26例);实验小白鼠60只,分健脾理肠片组及对照组,观察健脾理肠片对慢性结肠炎、溃疡性结肠炎患者免疫功能及对小白鼠巨噬细胞吞噬能力的影响。结果:50例病人的细胞免疫“E—玫瑰花试验,活性玫瑰花试验及淋巴细胞转化率”较治疗前有显著提高(P<0.01),IgA、IgM较治疗前显著降低(P<0.01),小白鼠巨噬细胞的吞噬百分率及吞噬指数明显提高(P<0.001),提示:健脾理肠片既能增强非特异性免疫功能,又能调节细胞和体液免疫的功能紊乱。

Clinical outcome of ileal pouch-anal anastomosis for chronic ulcerative colitis in China

Background The incidence of chronic ulcerative colitis （CUC） in China is remarkably increasing, while little information on surgical treatment has been reported. This study aimed to completely describe and analyze the clinical outcome of restorative proctocolectomy with ileal pouch-anal anastomosis （IPAA）for CUC in China. Methods Ninety-five consecutive patients, who suffered CUC and had surgical indications, were carefully selected. All patients underwent IPAA. Data on patient characteristics, surgical indications, surgical details, postoperative complications, functional outcome, and quality of life were collected. Results The mean patient age at the time of the operation was 32 years. Twenty-nine （31%） patients underwent an emergency operation, and 66 （69%） underwent elective procedures. Four patients with severe dysplasia underwent operations, but no carcinoma was histologically confirmed. A two-stage operation was performed in 87 （92%） patients, and a hand-sewn technique was applied in 88 （93%） patients. Sixteen patients （17.0%） experienced early complications, and there was a significant difference between the emergency surgery group and the elective group （31.0% vs. 10.6%, respectively; P 〈0.01）. Five （5.3%） patients developed pouchitis as a late complication. The mean stool frequency after the operation was 4.6 （2-11） during the first 24 hours and 1.5 （0-4） overnight. According to the Kirwan grading scale, 87 （91.8%） patients showed satisfactory anal continence function. The quality of life improved significantly from a preoperative mean value of 0.28-0.61 before ileostomy closure to 0.78 after ileostomy closure （P 〈0.01） according to the Cleveland Global Quality of Life index. Conclusions IPAA is an effective and safe surgical procedure for patients with CUC in China. However, some characteristics, such as the low incidence of pouchitis, require further study.