Clinical significance of the determination of fecal short-chain fatty acids in patients with nonalcoholic fatty liver disease / 临床肝胆病杂志

Objective To investigate the association of the metabolism of intestinal short-chain fatty acids (SCFAs) with the development and progression of the disease spectrum of nonalcoholic fatty liver disease (NAFLD) by determining the content of fecal SCFAs in patients with different NAFLD diseases and the change in the content of fecal SCFAs after treatment in patients at a high risk of nonalcoholic steatohepatitis (NASH). Methods A total of 90 patients who were diagnosed with NAFLD in The Affiliated Hospital of Qinghai University from July 2020 to July 2021 were enrolled and divided into simple nonalcoholic fatty liver (NAFL) group with 30 patients, NASH group with 30 patients, and nonalcoholic fatty liver fibrosis group with 30 patients, and 40 individuals who underwent physical examination during the same period of time were enrolled as control group. Related case data and fecal SCFAs content were collected for the four groups, and related clinical indices and fecal SCFAs content were collected for 10 patients at a high risk of NASH after 3 months of intervention. The analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the paired samples t -test was used for comparison within each group; the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the paired samples Wilcoxon signed rank sum test was used for comparison within each group; a Spearman correlation analysis was used to investigate the correlation between variables; the receiver operating characteristic (ROC) curve analysis was used for diagnostic evaluation. Results Compared with the control group, the nonalcoholic fatty liver fibrosis group had significantly higher contents of valeric acid and caproic acid, and the NAFL group had significantly lower contents of valeric acid and caproic acid (all P < 0.05); the nonalcoholic fatty liver fibrosis group had significantly higher contents of valeric acid and caproic acid than the NAFL group ( P < 0.05); the nonalcoholic fatty liver fibrosis group had a significantly higher content of valeric acid than the NASH group ( P < 0.05); the NASH group had a significantly higher content of caproic acid than the NAFL group ( P < 0.05). After treatment, the high-risk patients in the NASH group had significant reductions in HbA1c, fasting plasma glucose (FPG), triglyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), total bile acid (TBA), prothrombin time (PT), uric acid (UA), controlled attenuation parameter (CAP), and liver stiffness measurement (LSM) ( Z =-2.805, -2.703, -2.193, -2.599, -2.805, -2.701, -2.803, -1.988, -2.807, -2.803, -2.803, and -2.668, all P < 0.05); for these patients, the contents of acetic acid and propionic acid after treatment were significantly higher than those before treatment ( Z =-2.803 and -2.803, both P < 0.05), and the content of isobutyric acid after treatment was significantly lower than that before treatment ( Z =-2.803, P < 0.05). In the diagnosis of nonalcoholic fatty liver fibrosis, valeric acid had an area under the ROC curve (AUC) of 0.842, with a sensitivity of 86.7% and a specificity of 70% at the optimal cut-off value of 141.42 μg/g; caproic acid had an AUC of 0.819, with a sensitivity of 70% and a specificity of 85% at the optimal cut-off value of 6.93 μg/g. Conclusion Valeric acid and caproic acid may promote the development of NAFLD disease spectrum. Acetic acid and propionic acid may have a certain protective effect on the liver of NAFLD patients, and isobutyric acid may promote the development and progression of NASH. The protective effect of acetic acid and propionic acid on the liver may further lead to the reductions in HbA1c, FPG, TG, TC, ALT, AST, GGT, TBA, PT, UA, CAP, and LSM. Valeric acid and caproic acid have an inferior diagnostic value to PIIIP N-P and a superior diagnostic value to type IV collagen and hyaluronic acid. Valeric acid with the optimal cut-off value of 141.42 μg/g and caproic acid with the optimal cut-off value of 6.93 μg/g can be used as the auxiliary diagnostic indicators for the early diagnosis of nonalcoholic fatty liver fibrosis.

Effects of acupuncture on nutritional status in patients in a persistent vegetative state: a prospective randomized controlled study / 针灸推拿医学（英文版）

Objective: To explore the effects of acupuncture on nutritional status in patients in a persistent vegetative state. Methods: A prospective randomized controlled trial was designed. A total of 66 patients in a persistent vegetative state were randomized into a control group and an observation group, with 33 cases in each group. The control group was given conventional treatment plus enteral nutrition support. The observation group was treated with additional Tiao Shen Jian Pi acupuncture therapy (acupuncture for spirit-regulating and spleen-invigorating) based on the same interventions in the control group. Both groups were treated for 8 weeks. The levels of total protein (TP), prealbumin (PA), albumin (Alb), and hemoglobin (Hb) were measured before and after treatment. The upper arm circumference and skinfold thickness of triceps brachii were measured. And the intestinal flora and fecal short-chain fatty acids contents were determined.Results: After treatment, the levels of TP, PA, Alb, and Hb in the control group were decreased (P<0.05), while in the observation group, compared with those before treatment, the levels of TP, PA, Alb, and Hb had no statistical differences (P>0.05), and the levels were all higher than those in the control group (P<0.05). The upper arm circumference and skinfold thickness of triceps brachii in both groups decreased (P<0.05), and the values of these two items in the observation group were higher than those in the control group (P<0.05). In the control group, the contents of Bifidobacterium and Lactobacillus in feces decreased (P<0.05), and the content of Enterococcus increased (P<0.05). In the observation group, the contents of Bifidobacterium and Lactobacillus in feces increased (P<0.05), and the content of Enterococcus decreased (P<0.05). The differences between the two groups were statistically significant (P<0.05). In the control group, the total content of fecal short-chain fatty acids and the contents of acetic acid and butyric acid in feces decreased (P<0.05). In the observation group, the total content of fecal short-chain fatty acids and the contents of acetic acid and butyric acid in feces increased (P<0.05) and were all higher than those in the control group (P<0.05). Conclusion: Acupuncture can improve nutrition-related blood indicators in patients in a persistent vegetative state and delay the decrease of upper arm circumference and skinfold thickness of triceps brachii, which may be related to the regulation of intestinal flora and fecal short-chain fatty acids contents.

Correlation between gut microbiota, short-chain fatty acids metabolic disorder and outcomes of patients with hypertensive intracerebral hemorrhage / 国际脑血管病杂志

Objective:To investigate the metabolic disorder of gut microbiota and short-chain fatty acids (SCFAs) in patients with hypertensive intracerebral hemorrhage and their correlations with the poor outcomes.Methods:Thirty-eight patients with hypertensive intracerebral hemorrhage within 7 d of onset and 32 healthy controls were enrolled prospectively. Fecal samples were collected for 16S rRNA sequencing and SCFAs levels detection. The outcome was evaluated by the modified Rankin Scale at 90 d after the onset, and >2 points were defined as a poor outcome. Multivariate logistic regression model was used to determine the correlations between the gut microbiota and the fecal SCFAs levels and outcomes. Results:The gut microbiota of patients with hypertensive intracerebral hemorrhage was significantly different from that of healthy control group. It is manifested as a decrease in α diversity, a difference in β diversity, an increase in the abundance of potential undesirable bacteria, a decrease in the abundance of common SCFA-producing bacteria and a decrease in the fecal SCFAs levels. In patients with hypertensive intracerebral hemorrhage, compared with the good outcome group, the α diversity of the gut microbiota, the abundance of SCFA-producing bacteria such as Lacetospirillum and Bacteroides, and the total SCFAs, acetic acid and propionic acid levels decreased in the poor outcome group. Multivariate logistic regression analysis showed that after adjusting for potential confounding factors, the decrease of fecal SCFAs levels after log2 conversion was significantly and independently correlated with the poor outcomes. Conclusion:Patients with hypertensive intracerebral hemorrhage have gut microbiota and SCFAs metabolic disorder, the latter is significantly correlated with the poor outcomes. Gut microbiota and SCFAs may become an outcome marker and treatment target for patients with hypertensive intracerebral hemorrhage

Sucralfate enemas reduce the oxidative tissue damage and preserves the contents of E-cadherin and Β-catenin in colonic mucosa without fecal stream

ABSTRACT Purpose: To evaluate the effects of sucralfate enemas in tissue contents of E-cadherin and ?-catenin in an experimental diversion colitis. Methods: Thirty-six male Wistar rats were submitted to a proximal colostomy and a distal mucous fistula. They were allocated into three groups: first group received daily saline enemas (2 mL/day) and the two other groups daily enemas with sucralfate at dosage of 1 or 2 g/kg/day, respectively. Six animals of each group were euthanized after two weeks and six animals after four weeks. The inflammation of the excluded mucosa was evaluated by histological analysis. The oxidative damage was quantified by measurement of malondialdehyde tissue levels. The expression of E-cadherin and ?-catenin was identified by immunohistochemistry, and its contents were quantified by computer-assisted image analysis. Results: Sucralfate enemas reduced inflammation in animals subjected to treatment with 2 g/kg/day by four weeks, and the levels of oxidative damage in mucosa without fecal stream irrespective of concentration and time of intervention. E-cadherin and ?-catenin content increased in segments without fecal stream in those animals subjected to treatment with sucralfate. Conclusions: Sucralfate reduces the inflammation and oxidative stress and increases the tissue content of E-cadherin and ?-catenin in colonic mucosa devoid to the fecal stream.

Differential Effects of Lactobacillus casei Strain Shirota on Patients With Constipation Regarding Stool Consistency in China

BACKGROUND/AIMS: Probiotics are expected to confer benefits on patients with constipation, but how probiotics act on constipated patients with variable stool consistencies remains unclear. We investigated the effect of Lactobacillus casei strain Shirota (LcS) on constipation-related symptoms, especially stool consistency, of constipated patients. METHODS: Constipated patients meeting the Rome III criteria were divided into 3 groups according to the Bristol Stool Form Scale (BSFS): hard (hard stool [HS], BSFS < 3), normal (normal stool [NS], ≤ 3 BSFS ≤ 4), and soft (soft stool [SS], 4 < BSFS ≤ 5) stools. Subjects in each group consumed a probiotic beverage containing 1010 colony-forming units of LcS daily for 28 days. RESULTS: LcS intervention significantly alleviated constipation-related symptoms and increased defecation frequency in all subjects. Four weeks of LcS supplementation softened the hard stools in HS, hardened the soft stools in SS, and did not alter the ideal stool consistency in NS. The short-chain fatty acid (SCFA) concentrations were highest in SS, followed by NS and HS. LcS intervention increased the stool SCFA levels in HS but reduced or did not alter the levels in NS and SS. LcS intervention increased the Pseudobutyrivibrio and Roseburia abundances in HS and decreased the Pseudobutyrivibrio abundance in SS. CONCLUSIONS: LcS supplementation improved the constipation-related symptoms in constipated subjects. Differences in baseline stool consistency could result in different anti-constipation effects of LcS intervention. LcS balanced the stool consistency—softened the HS and hardened the SS. These effects could be associated with modulation of the gut microbiota and SCFA production.

Enemas with mesalazine increase the tissue contents of mucins in the colonic mucosa devoid of fecal stream

Abstract Purpose: To evaluate the inflammatory reaction and measure the content of mucins, in the colonic mucosa without fecal stream submit to intervention with mesalazine. Methods: Twenty-four rats were submitted to a left colostomy and a distal mucous fistula and divided into two groups according to euthanasia to be performed two or four weeks. Each group was divided into two subgroups according daily application of enemas containing saline or mesalazine at 1.0 g/kg/day. Colitis was diagnosed by histological analysis and the inflammatory reaction by validated score. Acidic mucins and neutral mucins were determined with the alcian-blue and periodic acid of Schiff techniques, respectively. Sulfomucin and sialomucin were identified by high iron diamine-alcian blue technique. The tissue contents of mucins were quantified by computer-assisted image analysis. Mann-Whitney test was used to analyze the results establishing the level of significance of 5%. Results: Enemas with mesalazine in colonic segments without fecal stream decreased the inflammation score and increased the tissue content of all subtypes of mucins. The increase of tissue content of neutral, acid and sulfomucin was related to the time of intervention. Conclusion: Mesalazine enemas reduce the inflammatory process and preserve the content of mucins in colonic mucosa devoid of fecal stream.

Evaluation of enemas containing sucralfate in tissue content of muc-2 protein in experimental model of diversion colitis / Avaliação de enemas com sucralfato no conteúdo tecidual da proteina muc-2 em colite de exclusão experimental

ABSTRACT Background: The effects of topical application of sucralfate (SCF) on the tissue content of MUC-2 protein have not yet been evaluated in experimental models of diversion colitis. Aim: To measure the tissue content of MUC-2 protein in the colonic mucosa diverted from fecal stream submitted to the SCF intervention. Methods: Thirty-six rats underwent derivation of intestinal transit through proximal colostomy and distal mucous fistula. The animals were divided into three groups which were submitted application of enemas with saline, SCF 1 g/kg/day and SCF 2 g/kg/day. Each group was divided into two subgroups, according to euthanasia was done after two or four weeks. The colitis diagnosis was established by histopathological study and the inflammatory intensity was evaluated by previously validated scale. The MUC-2 protein was identified by immunohistochemistry and the tissue content was measured computerized morphometry). Results: The application of enemas with SCF in the concentration of 2 g/kg/day reduced inflammatory score of the segments that were diverted from fecal stream. The content of MUC-2 in diverted colon of the animals submitted to the intervention with SCF, independently of intervention period and the used concentration, was significantly greater than animals submitted to the application of enemas containing saline (p< 0.01). The content of MUC-2 after the intervention with SCF in the concentration of 2 g/kg/day was significantly higher when compared to the animals submitted to the application containing SCF at concentration of 1.0 g/kg/day (p<0.01). The tissue content of MUC-2 reached the highest values after intervention with SCF in the concentration of 2 g/kg/day for four weeks (p<0.01). Conclusion: The preventive application of enemas containing SCF reduces the inflammatory score and avoids the reduction of tissue content of MUC-2, suggesting that the substance is a valid therapeutic strategy to preserve the mucus layer that covers the intestinal epithelium.

RESUMO Racional: Os efeitos da aplicação tópica de sucralfato (SCF) no conteúdo tecidual da proteína mucina-2 (MUC-2) ainda não foram avaliados em modelos experimentais de colite de exclusão. Objetivo: Mensurar o conteúdo tecidual da proteína MUC-2 na mucosa cólica sem trânsito intestinal submetida à intervenção com SCF. Método: Trinta e seis ratos foram submetidos à derivação intestinal por colostomia proximal terminal e fístula mucosa distal. Foram divididos em três grupos segundo recebessem clisteres contendo solução fisiológica (SF), SCF 1 g/kg/dia e SCF 2 g/kg/dia. Cada grupo foi dividido em dois subgrupos, segundo a eutanásia ser realizada após duas ou quatro semanas. O diagnóstico de colite foi estabelecido por estudo histopatológico e a intensidade inflamatória foi avaliada por escala validada. A expressão tecidual da MUC-2 foi identificada por imunoistoquímica e seu conteúdo mensurado por morfometria computadorizada. Resultados: A aplicação de clisteres com SCF na concentração de 2 g/kg/dia reduziu a intensidade inflamatória no cólon sem trânsito fecal. O conteúdo tecidual de MUC-2 no cólon sem trânsito dos animais submetidos à intervenção com SCF, independente do tempo de intervenção e da concentração utilizada, foi maior quando comparado aos animais tratados com SF (p<0,01). O conteúdo de MUC-2 após a intervenção com SCF na concentração de 2 g/kg/dia foi maior quando comparado aos animais submetidos à intervenção com concentração menor (p<0,01). O conteúdo de MUC-2 foi maior após intervenção com SCF na concentração de 2 g/kg/dia por quatro semanas (p<0,01). Conclusão: A aplicação preventiva de clisteres com SCF reduz o grau de inflamação e preserva o conteúdo tecidual de MUC-2, em segmentos desprovidos de trânsito intestinal, mostrando-se uma estratégia terapêutica válida para preservar a camada de muco que recobre o epitélio intestinal.

[retracted article]: evaluation of enemas containing sucralfate in tissue content of muc-2 protein in experimental model of diversion colitis / [artigo retratado]: avaliação de enemas com sucralfato no conteúdo tecidual da proteina muc-2 em colite de exclusão experimental

ABSTRACT Background: The effects of topical application of sucralfate (SCF) on the tissue content of MUC-2 protein have not yet been evaluated in experimental models of diversion colitis. Aim: To measure the tissue content of MUC-2 protein in the colonic mucosa diverted from fecal stream submitted to the SCF intervention. Methods: Thirty-six rats underwent derivation of intestinal transit through proximal colostomy and distal mucous fistula. The animals were divided into three groups which were submitted application of enemas with saline, SCF 1 g/kg/day and SCF 2 g/kg/day. Each group was divided into two subgroups, according to euthanasia was done after two or four weeks. The colitis diagnosis was established by histopathological study and the inflammatory intensity was evaluated by previously validated scale. The MUC-2 protein was identified by immunohistochemistry and the tissue content was measured computerized morphometry). Results: The application of enemas with SCF in the concentration of 2 g/kg/day reduced inflammatory score of the segments that were diverted from fecal stream. The content of MUC-2 in diverted colon of the animals submitted to the intervention with SCF, independently of intervention period and the used concentration, was significantly greater than animals submitted to the application of enemas containing saline (p< 0.01). The content of MUC-2 after the intervention with SCF in the concentration of 2 g/kg/day was significantly higher when compared to the animals submitted to the application containing SCF at concentration of 1.0 g/kg/day (p<0.01). The tissue content of MUC-2 reached the highest values after intervention with SCF in the concentration of 2 g/kg/day for four weeks (p<0.01). Conclusion: The preventive application of enemas containing SCF reduces the inflammatory score and avoids the reduction of tissue content of MUC-2, suggesting that the substance is a valid therapeutic strategy to preserve the mucus layer that covers the intestinal epithelium.

RESUMO Racional: Os efeitos da aplicação tópica de sucralfato (SCF) no conteúdo tecidual da proteína mucina-2 (MUC-2) ainda não foram avaliados em modelos experimentais de colite de exclusão. Objetivo: Mensurar o conteúdo tecidual da proteína MUC-2 na mucosa cólica sem trânsito intestinal submetida à intervenção com SCF. Método: Trinta e seis ratos foram submetidos à derivação intestinal por colostomia proximal terminal e fístula mucosa distal. Foram divididos em três grupos segundo recebessem clisteres contendo solução fisiológica (SF), SCF 1 g/kg/dia e SCF 2 g/kg/dia. Cada grupo foi dividido em dois subgrupos, segundo a eutanásia ser realizada após duas ou quatro semanas. O diagnóstico de colite foi estabelecido por estudo histopatológico e a intensidade inflamatória foi avaliada por escala validada. A expressão tecidual da MUC-2 foi identificada por imunoistoquímica e seu conteúdo mensurado por morfometria computadorizada. Resultados: A aplicação de clisteres com SCF na concentração de 2 g/kg/dia reduziu a intensidade inflamatória no cólon sem trânsito fecal. O conteúdo tecidual de MUC-2 no cólon sem trânsito dos animais submetidos à intervenção com SCF, independente do tempo de intervenção e da concentração utilizada, foi maior quando comparado aos animais tratados com SF (p<0,01). O conteúdo de MUC-2 após a intervenção com SCF na concentração de 2 g/kg/dia foi maior quando comparado aos animais submetidos à intervenção com concentração menor (p<0,01). O conteúdo de MUC-2 foi maior após intervenção com SCF na concentração de 2 g/kg/dia por quatro semanas (p<0,01). Conclusão: A aplicação preventiva de clisteres com SCF reduz o grau de inflamação e preserva o conteúdo tecidual de MUC-2, em segmentos desprovidos de trânsito intestinal, mostrando-se uma estratégia terapêutica válida para preservar a camada de muco que recobre o epitélio intestinal.

Influence of the application of enemas with oily extract of curcumin (Curcuma longa) on the tissue content of neutral and acidic mucins in the colonic mucosa without fecal stream / Influência da aplicação de clisteres contendo extrato oleoso de curcumina (Curcuma longa) no conteúdo tecidual de mucinas neutras e ácidas na mucosa cólica sem trânsito intestinal

ABSTRACT Purpose: To verify if the application of enemas containing oily extracts of curcumin preserves the tissue content of mucins in the glands of the colonic mucosa without fecal stream. Method: Thirty-six Wistar rats were submitted to diversion of the fecal stream by proximal colostomy and distal mucous fistula. The animals were subdivided into three groups, and accordingly received enemas with saline and oily extract of curcumin at concentrations of 50 mg/kg/day or 200 mg/kg/day. After two or four weeks of intervention, the irrigated colic segments were removed. Neutral and acidic mucins were identified by Periodic-acid Schiff and Alcian-Blue techniques, respectively. The content of both mucin subtypes was measured by computerized morphometry. Mann-Whitney test was used to analyze the results, adopting a significance level of 5% (p ≤ 0.05). Results: There was an increase in the tissue content of neutral mucins in animals treated with curcumin at a concentration of 50 mg/kg/day for four weeks, whereas in the group treated with 200 mg/kg/day there was an increase independent of the time of intervention. The content of acidic mucins increased in animals treated with 200 mg/kg/day regardless of the intervention time, whereas in those treated with 50 mg/kg/day an increase was observed only after four weeks. Conclusion: Enemas with curcumin preserve the content of neutral and acidic mucins in the colonic epithelium without fecal stream.

RESUMO Objetivo: Verificar se a aplicação de clisteres com extrato oleoso de curcumina preserva o conteúdo de mucinas nas glândulas da mucosa cólica sem trânsito intestinal. Método: Trinta e seis ratos Wistar foram submetidos à derivação intestinal por colostomia proximal e fístula mucosa distal. Os animais foram subdivididos em três grupos, segundo receberem clisteres com soro fisiológico 0,9%, extrato oleoso de curcumina nas concentrações de 50 mg/kg/dia ou 200 mg/kg/dia. Após duas ou quatro semanas de intervenção foram removidos os segmentos cólicos irrigados. As mucinas neutras e ácidas foram identificadas pelas técnicas do PAS e Alcian-Blue, respectivamente. O conteúdo tecidual de ambos os subtipos de mucinas foi mensurado por morfometria computadorizada. Utilizou-se teste de Mann-Whitney para análise dos resultados adotando-se nível de significância de 5% (p ≤ 0,05). Resultados: Houve aumento no conteúdo de mucinas neutras nos animais tratados com curcumina na concentração de 50 mg/kg/dia por quatro semanas, enquanto nos tratados com 200 mg/kg/dia houve aumento independente do tempo de intervenção. O conteúdo de mucinas ácidas aumentou nos animais tratados com 200 mg/kg/dia independente do tempo de intervenção, enquanto nos tratados com 50 mg/kg/dia encontrou-se aumento apenas após quatro semanas. Conclusão: Clisteres com curcumina preservam o conteúdo de mucinas neutras e ácidas no epitélio cólico sem trânsito intestinal.

Evaluation of enemas containing sucralfate in tissue content of muc-2 protein in experimental model of diversion colitis / Avaliação de enemas com sucralfato no conteúdo tecidual da proteina muc-2 em colite de exclusão experimental

ABSTRACT Background: The effects of topical application of sucralfate (SCF) on the tissue content of MUC-2 protein have not yet been evaluated in experimental models of diversion colitis. Aim: To measure the tissue content of MUC-2 protein in the colonic mucosa diverted from fecal stream submitted to the SCF intervention. Methods: Thirty-six rats underwent derivation of intestinal transit through proximal colostomy and distal mucous fistula. The animals were divided into three groups which were submitted application of enemas with saline, SCF 1 g/kg/day and SCF 2 g/kg/day. Each group was divided into two subgroups, according to euthanasia was done after two or four weeks. The colitis diagnosis was established by histopathological study and the inflammatory intensity was evaluated by previously validated scale. The MUC-2 protein was identified by immunohistochemistry and the tissue content was measured computerized morphometry). Results: The application of enemas with SCF in the concentration of 2 g/kg/day reduced inflammatory score of the segments that were diverted from fecal stream. The content of MUC-2 in diverted colon of the animals submitted to the intervention with SCF, independently of intervention period and the used concentration, was significantly greater than animals submitted to the application of enemas containing saline (p< 0.01). The content of MUC-2 after the intervention with SCF in the concentration of 2 g/kg/day was significantly higher when compared to the animals submitted to the application containing SCF at concentration of 1.0 g/kg/day (p<0.01). The tissue content of MUC-2 reached the highest values after intervention with SCF in the concentration of 2 g/kg/day for four weeks (p<0.01). Conclusion: The preventive application of enemas containing SCF reduces the inflammatory score and avoids the reduction of tissue content of MUC-2, suggesting that the substance is a valid therapeutic strategy to preserve the mucus layer that covers the intestinal epithelium.

RESUMO Racional: Os efeitos da aplicação tópica de sucralfato (SCF) no conteúdo tecidual da proteína mucina-2 (MUC-2) ainda não foram avaliados em modelos experimentais de colite de exclusão. Objetivo: Mensurar o conteúdo tecidual da proteína MUC-2 na mucosa cólica sem trânsito intestinal submetida à intervenção com SCF. Método : Trinta e seis ratos foram submetidos à derivação intestinal por colostomia proximal terminal e fístula mucosa distal. Foram divididos em três grupos segundo recebessem clisteres contendo solução fisiológica (SF), SCF 1 g/kg/dia e SCF 2 g/kg/dia. Cada grupo foi dividido em dois subgrupos, segundo a eutanásia ser realizada após duas ou quatro semanas. O diagnóstico de colite foi estabelecido por estudo histopatológico e a intensidade inflamatória foi avaliada por escala validada. A expressão tecidual da MUC-2 foi identificada por imunoistoquímica e seu conteúdo mensurado por morfometria computadorizada. Resultados: A aplicação de clisteres com SCF na concentração de 2 g/kg/dia reduziu a intensidade inflamatória no cólon sem trânsito fecal. O conteúdo tecidual de MUC-2 no cólon sem trânsito dos animais submetidos à intervenção com SCF, independente do tempo de intervenção e da concentração utilizada, foi maior quando comparado aos animais tratados com SF (p<0,01). O conteúdo de MUC-2 após a intervenção com SCF na concentração de 2 g/kg/dia foi maior quando comparado aos animais submetidos à intervenção com concentração menor (p<0,01). O conteúdo de MUC-2 foi maior após intervenção com SCF na concentração de 2 g/kg/dia por quatro semanas (p<0,01). Conclusão: A aplicação preventiva de clisteres com SCF reduz o grau de inflamação e preserva o conteúdo tecidual de MUC-2, em segmentos desprovidos de trânsito intestinal, mostrando-se uma estratégia terapêutica válida para preservar a camada de muco que recobre o epitélio intestinal.

Evaluation of the application of enemas containing sucralfate in tissue content of neutral and acid mucins in experimental model of diversion colitis

PURPOSE: To evaluate the effects of sucralfate on tissue content of neutral and acids mucins in rats with diversion colitis. METHODS: Thirty-six rats were submitted to a proximal right colostomy and a distal mucous fistula. They were divided into two groups according to sacrifice to be performed two or four weeks after intervention. Each group was divided into three subgroups according daily application of enemas containing saline, sucralfate at 1.0 g/kg/day or 2.0 g/kg/day. Colitis was diagnosed by histological analysis and neutral and acid mucins by Periodic Acid Schiff and Alcian Blue techniques, respectively. The contents of mucins were quantified by computer-assisted image analysis. Student's t paired and ANOVA test were used to compare the contents of both types of mucins among groups, and to verify the variance with time, establishing level of signification of 5% for both (p<0.05). RESULTS: Enemas containing sucralfate improves the inflammation and increases the tissue contents of neutral and acid mucins. The content of neutral mucins does not change with the time or concentration of sucralfate used, while acid mucins increases with concentration and time of intervention. CONCLUSIONS: Sucralfate enemas improve the inflammatory process and increase the tissue content of neutral and acid mucins in colon without fecal stream. .

Oxidative stress and changes in the content and pattern of tissue expression of ß-catenin protein in diversion colitis

OBJECTIVE: The aim of this study is to verify if oxidative stress is related to changes in content and pattern of β-catenin protein expression in an experimental model of diversion colitis. METHODS: Sixty Wistar rats were submitted to intestinal bypass. The animals were divided into three groups according to the sacrifice to take place in six, 12 and 18 weeks. For each group, five animals only underwent laparotomy (control). The presence of colitis was diagnosed by histological study, and its severity, by inflammation grading scale. Cellular oxidative stress was measured by comet assay. Tissue expression of β-catenin protein was analyzed by the immunohistochemistry and quantification of its tissue content by computerized morphometry. Statistical analysis was performed with the Student's t-test, median, Mann-Whitney, ANOVA and Kruskal-Wallis, adopting a significance level of 5% (p <0.05). RESULTS: Colon segments without fecal stream developed colitis, which worsened with time of exclusion. Segments without fecal stream suffer higher levels of oxidative stress when compared to those with stream, and it worsens with time of exclusion. The levels of cellular oxidative stress are directly related to the degree of inflammation. The total content of ß-catenin in segments without fecal stream reduces after six weeks, and does not vary thereafter. The content of β-catenin in the apical portion of the colon crypts decreases with time, whereas in the basal region, it increases. The total content of ß-catenin is inversely related to the degree of inflammation and levels of tissue oxidative stress levels. CONCLUSION: There are changes in tissue content of E-cadherin and increased expression of ß-catenin in proliferative regions of colonic crypts, related with oxidative tissue stress. (AU)

OBJETIVO: O objetivo do presente estudo é avaliar a relação entre estresse oxidativo e conteúdo tecidual de β-catenina em modelo experimental de colite de exclusão. MÉTODOS: Sessenta ratos Wistar foram submetidos à derivação intestinal e divididos em três grupos experimentais segundo o sacrifício ser realizado em 6, 12 e 18 semanas. Para cada grupo, cinco animais foram submetidos apenas a laparotomia (controle). A colite foi diagnosticada por estudo histológico, enquanto sua intensidade por escala de graduação inflamatória. Os níveis de estresse oxidativo foram mensurados pelo ensaio cometa, enquanto a expressão e o conteúdo tecidual de ß-catenina por imunoistoquímica e morfometria computadorizada, respectivamente. Os resultados foram analisados pelos testes t de Student, Mann Whitney, ANOVA e Kruskal-Wallis, estabelecendo-se nível de significância de 5% (p<0,05). RESULTADOS: Nos segmentos sem trânsito fecal ocorre desenvolvimento de colite que piora com o tempo de exclusão. Segmentos sem trânsito sofrem maiores níveis de estresse oxidativo quando comparados àqueles com trânsito, piorando com o tempo de exclusão. Os níveis de estresse oxidativo encontram-se diretamente relacionados a piora da inflamação. O conteúdo total de ß-catenina no cólon sem trânsito reduz após seis semanas de exclusão. O conteúdo de ß-catenina no ápice das criptas cólicas diminui com o tempo, enquanto na região basal, aumenta. O conteúdo total da β-catenina encontra-se inversamente relacionado ao grau de inflamação e aos níveis de estresse oxidativo. CONCLUSÃO: Existe redução no conteúdo de ß-catenina, principalmente no ápice das glândulas cólicas e aumento nas regiões basais, relacionadas à piora do estresse oxidativo. (AU)

Avaliação do número de células caliciformes nas criptas da mucosa colônica com e sem trânsito intestinal / Evaluation of the number of goblet cells in crypts of the colonic mucosa with and without fecal transit

OBJETIVO: Medir a espessura das criptas e quantificar o número de células caliciformes comparando a mucosa cólica com e sem trânsito intestinal, relacionando-as ao tempo de exclusão. MÉTODOS: Sessenta ratos Wistar, foram distribuídos em três grupos com 20 animais segundo a operação final para a retirada dos cólons, realizadas em seis, 12 ou 18 semanas. Em cada grupo, 15 animais foram submetidos à derivação do trânsito por colostomia proximal no cólon esquerdo e fístula mucosa distal e cinco apenas à laparotomia (controle). Os cólons com e sem trânsito fecal foram removidos, processados, submetidos a cortes histológicos corados pela hematoxilina-eosina. A altura das criptas colônicas e o número de células caliciformes foram mensurados por morfometria computadorizada. Foram utilizados os testes t de Student e Kruskal-Wallis para comparação e análise de variância, estabelecendo-se nível de significância de 5% (p<0,05). RESULTADOS: A altura das criptas diminui nos segmentos sem trânsito fecal (p=0,0001), reduzindo entre seis e 12 semanas de exclusão (p=0,0003), estabilizando-se após este período. O número de células caliciformes nas criptas é menor nos segmentos sem trânsito após 12 e 18 semanas (p=0,0001), porém aumenta com o decorrer do tempo de exclusão (p=0,04) CONCLUSÃO: A exclusão do trânsito intestinal diminui a espessura das criptas colônicas e o número de células caliciformes nos segmentos sem trânsito. Existe aumento do número de células caliciformes com o decorrer do tempo de exclusão.

OBJECTIVE: To measure the thickness of the crypts and quantify the number of goblet cells of the colonic mucosa with and without intestinal transit, relating them to exclusion time. METHODS: Sixty Wistar rats were divided into three groups of 20 animals each according to the time of the final operation for the removal of the colon, in six, 12 or 18 weeks. In each group 15 animals underwent colonic exclusion by left colon proximal colostomy and distal mucous fistula, and five underwent only laparotomy (control). The colons with and without fecal stream were removed, processed and submitted to histological sections stained with hematoxylin-eosin. The height of the colonic crypts and the number of goblet cells were measured by computerized morphometry. We used the Student t test and Kruskal-Wallis test for comparison and analysis of variance, using a significance level of 5% (p <0.05). RESULTS: The height of the crypts decreased in segments without fecal stream (p =0.0001), reducing from six to 12 weeks of exclusion (p = 0.0003), stabilizing thereafter. The number of goblet cells in the crypts was smaller in segments without transit after 12 and 18 weeks (p = 0.0001), but increased as the time of exclusion progressed (p = 0.04) CONCLUSION: The exclusion of intestinal transit decreases the thickness of the colonic crypts and the number of goblet cells in the segments without transit. There is an increased number of goblet cells in the course of time exclusion.

Efeitos do butirato nos níveis de peroxidação lipídica em células da mucosa cólica sem trânsito fecal: estudo experimental em ratos / Effects of butyrate on levels of lipid peroxidation in cells of the colonic mucosa without fecal stream: experimental study in rats

Os ácidos graxos de cadeia curta (AGCC) representam o principal substrato energético para células da mucosa cólica. A derivação intestinal, reduzindo suprimento de AGCC, responsabiliza-se pela colite de exclusão (CE). Aplicação retal de butirato tem sido eficaz no tratamento da doença. Então, o objetivo deste estudo foi avaliar os níveis de lipoperoxidação na mucosa cólica, após aplicação de butirato, em modelo de CE. Vinte seis ratos Wistar foram submetidos à colostomia proximal e fístula mucosa distal. Os animais foram divididos em dois grupos segundo sacrifício ser realizado em duas ou quatro semanas. Cada grupo foi subdividido em dois subgrups segundo intervenção com soro fisiológico ou butirato. O diagnóstico de CE foi estabelecido por estudo histopatológico e os níveis de lipoperoxidação pelos níveis de malondialdeído (MDA). Utilizaram-se os testes de Mann-Whitney e Kruskal-Wallis (significantes quando p<0,05). Após duas semanas, os níveis de MDA foram menores nos segmentos sem trânsito nos animais irrigados com butirato (p=0,006); porém, após quatro semanas foram semelhantes (p=0,08). No cólon sem trânsito irrigado com butirato, os níveis de MDA aumentaram com o tempo de exclusão (p=0,02); enquanto no cólon com trânsito não se modificaram (p=0,86). O butirato reduz os níveis de MDA na mucosa cólica sem trânsito fecal, após duas semanas de derivação; entretanto, a irrigação isolada não é capaz de reduzir os níveis de lipoperoxidação das células mucosas com o progredir do tempo de exclusão intestinal.

The short-chain fatty acids (SCFA) are the main energy substrate for the cells of the colonic mucosa. Diversion of the fecal stream reducing the supply of SCFA is responsible for diversion colitis (DC). Rectal application of butyrate has been demonstrated effective in the treatment of the disease. So the aim of this study was to evaluate the levels of lipid peroxidation in the colon mucosa after application of butyrate in model of DC. Twenty-six rats were submitted to proximal colostomy and distal mucous fistula. The animals were divided into two groups according sacrifice carried out in two or four weeks. Each group was divided into two subgroups according to intervention with saline solution or butyrate. The diagnosis of colitis was established by histopathology and the levels of lipid peroxidation by tissue levels of malondialdehyde (MDA). We used the Mann-Whitney and Kruskal-Wallis, establishing a significance level of 5 percent (significant with p<0.05). After two weeks, the levels of MDA were lower in the segments without fecal stream of animals irrigated with butyrate (p=0.006), but after four weeks were similar (p=0.08). In the colon without fecal stream irrigated with butyrate, MDA levels increased with the time (p=0.02), while in segments with fecal stream MDA not changed. (p=0.86). Butyrate reduces the levels of MDA in the colonic mucosa without fecal stream, after two weeks of derivation. However, the irrigation with the substance is not able to reduce the lipid peroxidation of mucosal cells with increasing time of intestinal exclusion.

The importance of oxygen free radicals in the etiopathogenesis of diversion colitis in rats / A importância dos radicais livres de oxigênio na etiopatogenia da colite de exclusão em ratos

PURPOSE: Quantify the levels of oxidative DNA damage of epithelial colon cells comparing segments with and without fecal stream. METHODS: Sixty Wistar rats were subjected to deviation of fecal stream by proximal colostomy and a distal mucosal fistula. Animals were divided into three experimental groups that were sacrificed 6, 12 and 24 weeks after surgery. In each experimental group, five animals underwent laparotomy without intestinal deviation (sham subgroup). The diagnosis of colitis was made by histopathological analysis and the inflammatory activity index by graduated scale. The neutrophil infiltration was determined by myeloperoxidase tissue levels and the intensity of oxidative DNA damage by comet assay. The Mann-Withney and Student t test were used to compare the results among experimental subgroups and the Kruskal-Wallis test for variance analysis, adopting a significance level of 5 percent (p<0.05). RESULTS: Colon segments without fecal stream was shown higher histological inflammatory score of the colon wall after 12 and 24 weeks (p=0.001) that increased with the time of diversion (p=0.01). The activity of myeloperoxidase in segments without fecal stream decreased with the time (p=0.001). Oxidative DNA damage levels were significantly higher in the segments without fecal stream, (p=0.0001), independent of time of colon diversion, and increase with the time (p=0.0007). CONCLUSIONS: Colon segments without fecal stream showed high levels of oxidative DNA damage related to histological alterations observed in diversion colitis. The levels of oxidative DNA damage in segments devoid of the fecal stream increase with the time of intestinal exclusion.

OBJETIVO: Quantificar os níveis de dano oxidativo ao DNA em células epiteliais da mucosa cólica comparando segmentos com e sem trânsito fecal. MÉTODOS: Sessenta ratos Wistar foram submetidos à derivação do trânsito intestinal por colostomia proximal e fístula mucosa distal. Os animais foram divididos em três grupos experimentais segundo terem sido sacrificados 6, 12 e 24 semanas após a cirurgia. Em cada grupo experimental, cinco animais foram submetidos à laparotomia isolada sem derivação fecal (grupo sham). O diagnóstico de colite foi estabelecido por análise histopatológica e o índice de atividade inflamatória por escala graduada. A infiltração neutrofílica foi determinada pelos níveis teciduais da mieloperoxidase e a intensidade do dano oxidativo ao DNA pelo ensaio em cometa. Utilizaram-se os testes de Mann-Withney e o teste t de Student para comparar os resultados encontrados entre os subgrupos experimentais e o teste de Kruskal-Wallis para análise de variância, adotando-se nível de significância de 5 por cento (p<0,05). RESULTADOS: Os segmentos cólicos, sem trânsito fecal apresentaram maior escore histológico de inflamação após 12 e 24 semanas (p=0,001), que aumentou com o tempo de derivação (p=0,01). A atividade da mieloperoxidase nos segmentos sem trânsito fecal diminuiu com o progredir do tempo (p=0,001). Os níveis de dano oxidativo ao DNA foram significativamente maiores nos segmentos sem trânsito fecal (p=0,0001), independente do tempo de exclusão considerado, aumentando com o progredir do tempo de exclusão (p = 0,0007). CONCLUSÕES: Segmentos cólicos desprovidos de trânsito fecal apresentam níveis elevados de dano oxidativo ao DNA relacionados às alterações histológicas observadas na colite de exclusão. Os níveis de dano oxidativo ao DNA nos segmentos desprovidos de trânsito fecal aumentam com o decorrer do tempo de exclusão.

Avaliação dos níveis de peroxidação lipídica em células da mucosa cólica após aplicação de enemas com peróxido de hidrogênio: estudo experimental em ratos / Evaluation of lipid peroxidation levels on mucosa colonic cells afther application of hydrogen peroxide in enemas: experimental study in rats

A aplicação de clisteres contendo peróxido de hidrogênio (H2O2) determina o aparecimento de quadros graves de colite, algumas vezes de evolução fatal. É possível que a colite induzida por H2O2 possa ocorrer pela quebra da barreira funcional do epitélio cólico por estresse oxidativo. Objetivo: Avaliar os níveis de peroxidação lipídica em células da mucosa cólica após instilação de H2O2 no reto excluso de trânsito fecal. Método: Vinte seis ratos Wistar machos foram submetidos a colostomia proximal terminal no cólon descendente e fístula mucosa distal. Os animais foram randomizados em dois grupos segundo o sacrifício ter sido realizado duas ou quatro semanas após a derivação intestinal. Cada grupo experimental foi dividido e dois subgrupos segundo aplicação de clisteres, em dias alternados, contendo solução fisiológica a 0,9 por cento ou H2O2 a 3 por cento. O diagnóstico de colite foi estabelecido por estudo histopatológico e os níveis de dano oxidativo tecidual pela dosagem de malondialdeído por espectrofotometria. Os resultados foram analisados com os testes de Mann-Whitney e Kruskal-Wallis, estabelecendo-se nível de significância de 5 por cento (p menor que0,05)

Resultados: Os níveis de malondialdeído nos irrigados com SF nos cólons com e sem trânsito fecal após duas e quatro semanas de irrigação foram de: 0,05 mais ou menos 0,006; 0,06 mais ou menos 0,006 e 0,05 mais ou menos 0,03, 0,08 mais ou menos 0,02, respectivamente. Os níveis de malondialdeído nos irrigados com H2O2, nos cólons com e sem trânsito, após duas e quatro semanas de irrigação foram de 0,070 mais ou menos 0,006; 0,077 mais ou menos 0,01 e 0,052 mais ou menos 0,01, 0,08 mais ou menos 0,04, respectivamente. Após duas semanas os níveis de malondialdeído foram maiores nos animais irrigados com H2O2 em relação ao grupo controle (p= 0,007 e p= 0,01, respectivamente). Após quatro semanas não houve diferenças significantes Não ocorreu variação nos níveis de malondialdeído com o decorrer tempo de irrigação. Conclusão: Clisteres com H2O2, podem determinar o aparecimento de colite por ocasionarem estresse oxidativo nas células epiteliais da mucosa intestinal.

The use of rectal enemas with hydrogen peroxide (H2O2) determines the onset of severe colitis, sometimes with fatal evolution. It is possible that H2O2-induced colitis can occur by damage to the functional epithelial barrier of the colon by oxidative stress. Objective: The aim of present study was evaluate the levels of lipid peroxidation in cells of the colonic mucosa after instillation of H2O2 into the rectum excluded from fecal transit. Method: Twenty six male Wistar rats were undergone to proximal terminal colostomy in the descending colon and distal mucous fistula. The animals were randomized in two experimental groups according to the sacrifice was made two or four weeks after diversion of the fecal stream. Each experimental group was divided into two subgroups second application of enemas containing saline solution 0.9 percent or 3 percent H2O2 on alternate days. The diagnosis of colitis was established by histopathology study and the oxidative damage by tissue levels of malondialdehyde quantified by spectrophotometry. The results were analyzed with the Mann-Whitney and Kruskal-Wallis test, adopting a significance level of 5 percent (p less than 0.05).

Results: The levels of malondialdehyde in colon segments irrigated with saline, with and without fecal stream after two and four weeks of irrigation were: 0.05 more or less 0.006, 0.06 more or less 0.006 and 0.05 more or less 0.03, 0.08 more or less 0.02, respectively. The levels of malondialdehyde in colon segments irrigated with H2O2, in the colon with and without fecal stream, after two and four weeks of irrigation were 0.070 more or less 0.006, 0.077 and 0.052 more or less 0.01 more or less 0.01, 0.08 more or less 0.04, respectively . After two weeks the levels of malondialdehyde were higher on animals irrigated with H2O2 than control group (p = 0.007 and p = 0.01, respectively). After four weeks there were no significant differences in malondialdehyde levels related with the time of irrigation. Conclusion: Rectal enemas with H2O2, may determine the onset of colitis by oxidative stress on epithelial cells of intestinal mucosa.