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1.
BMC Complement Med Ther ; 22(1): 129, 2022 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-35534842

RESUMO

BACKGROUND: Jianpi Qinghua Fomula (JPQHF), a clinically proven prescription,has been applied to cure insulin resistance(IR) and type 2 diabetes (T2DM) for more than 20 years. Here, we will unravel the underlying molecular mechanisms relevant to the therapeutic actions of JPQHF. METHODS: High-fat(HF)diet-induced obesity(DIO)mouse were established in our research, along with insulin resistance. After the administration of JPQHF 5 or 6 weeks, the parameters of the glucose and lipid metabolism were measured. Flow cytometry and Luminex were utilized to assess the inflammation in small intestine,whilst Western blot was used to determine the relative expression levels of the MAPK pathway-related proteins. The glucose and lipid transporter of small intestine was assessed by immunofluorescence and ELISA, and the expression of insulin signaling pathway was detected by Western blot. RESULTS: The metabolic phenotypes of DIO mouse were ameliorated after 6-week oral administration of JPQHF; Meanwhile,JPQHF downregulated levels of IL-1ß,IL-6, TNF-α and IFN-γ but upregulated the ratio of M2/M1 macrophages in the small intestine. The elevated expressions of p-P38 MAPK/P38 MAPK、p-JNK/JNK and p-ERK1/2/ERK1/2 were reversed by JPQHF. Moreover, JPQHF enhanced expression of PI3K,p-AKT/AKT, p-IRS1/ IRS1, p-IRS2/ IRS2 and apoB48 in small intestine, and facilitated the translocation of GLUT2 to the basal side of small intestine epithelial cells. CONCLUSION: JPQHF alleviates insulin resistance in DIO mice, and this effect may be associated with its restraining of inflammation of small intestine via attenuating MAPK pathway, and then diminishes small intestinal glucose and lipid absorption.


Assuntos
Diabetes Mellitus Tipo 2 , Resistência à Insulina , Animais , Diabetes Mellitus Tipo 2/tratamento farmacológico , Glucose/metabolismo , Inflamação/tratamento farmacológico , Intestino Delgado/metabolismo , Lipídeos , Camundongos , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
2.
Physiol Rep ; 10(7): e15247, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35385223

RESUMO

In the present study, we examined the systemic and direct effects of parathyroid hormone (PTH) and fibroblast growth factor-23 (FGF-23) on duodenal, jejunal, and ileal Mg2+ absorption. The rats were injected with FGF-23 or PTH for 5 h before collecting the duodenum, jejunum, and ileum for Mg2+ transport analysis in Ussing chambers. The duodenum, jejunum, and ileum were directly exposed to FGF-23, PTH, or FGF-23 plus PTH with or without cell signaling inhibitors for 150 min in Ussing chambers prior to performing the Mg2+ transport study. The small intestinal tissues were also subjected to western blot analyses for FGF receptor (FGFR), PTH receptor (PTHR), Klotho, transient receptor potential melastatin 6 (TRPM6), and cyclin as well as the cystathionine ß-synthase domain divalent metal cation transport mediator 4 (CNNM4) expression. The small intestine abundantly expressed FGFR and PTHR proteins, whereas, Klotho was not expressed in rat small intestine. Systemic PTH or FGF-23 injection significantly suppressed transcellular Mg2+ transport in the duodenum and jejunum. Direct FGF-23-, PTH-, or FGF-23 plus PTH exposure also suppressed transcellular Mg2+ absorption in the duodenum and jejunum. There was no additional inhibitory effect of PTH and FGF-23 on intestinal Mg2+ absorption. The inhibitory effect of PTH, FGF-23, or FGF-23 plus PTH was abolished by Gö 6850. Systemic PTH- or FGF-23-injection significantly decreased membranous TRPM6 expression, but increased cytosolic CNNM4 expression in the duodenum, jejunum, and ileum. In the present study, we propose a novel magnesiotropic action of PTH and FGF-23 by modulating small intestinal Mg2+ absorption.


Assuntos
Proteínas de Transporte de Cátions , Hormônio Paratireóideo , Animais , Proteínas de Transporte de Cátions/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Absorção Intestinal , Intestino Delgado/metabolismo , Magnésio/metabolismo , Hormônio Paratireóideo/metabolismo , Hormônio Paratireóideo/farmacologia , Ratos
3.
Nutrients ; 14(8)2022 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-35458125

RESUMO

Vitamin A (VA) deficiency and diarrheal diseases are both serious public health issues worldwide. VA deficiency is associated with impaired intestinal barrier function and increased risk of mucosal infection-related mortality. The bioactive form of VA, retinoic acid, is a well-known regulator of mucosal integrity. Using Citrobacter rodentium-infected mice as a model for diarrheal diseases in humans, previous studies showed that VA-deficient (VAD) mice failed to clear C. rodentium as compared to their VA-sufficient (VAS) counterparts. However, the distinct intestinal gene responses that are dependent on the host's VA status still need to be discovered. The mRNAs extracted from the small intestine (SI) and the colon were sequenced and analyzed on three levels: differential gene expression, enrichment, and co-expression. C. rodentium infection interacted differentially with VA status to alter colon gene expression. Novel functional categories downregulated by this pathogen were identified, highlighted by genes related to the metabolism of VA, vitamin D, and ion transport, including improper upregulation of Cl- secretion and disrupted HCO3- metabolism. Our results suggest that derangement of micronutrient metabolism and ion transport, together with the compromised immune responses in VAD hosts, may be responsible for the higher mortality to C. rodentium under conditions of inadequate VA.


Assuntos
Infecções por Enterobacteriaceae , Deficiência de Vitamina A , Animais , Citrobacter rodentium , Colo/metabolismo , Diarreia/complicações , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Vitamina A/metabolismo , Deficiência de Vitamina A/complicações
4.
Cell Rep ; 38(9): 110438, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35235783

RESUMO

Intestinal epithelial cells derive from stem cells at the crypt base and travel along the crypt-villus axis to die at the villus tip. The two dominant villus epithelial cell types, absorptive enterocytes and mucous-secreting goblet cells, are mature when they exit crypts. Murine enterocytes switch functional cell states during migration along the villus. Here, we ask whether this zonation is driven by the bone morphogenetic protein (BMP) gradient, which increases toward the villus. Using human intestinal organoids, we show that BMP signaling controls the expression of zonated genes in enterocytes. We find that goblet cells display similar zonation involving antimicrobial genes. Using an inducible Bmpr1a knockout mouse model, we confirm that BMP controls these zonated genes in vivo. Our findings imply that local manipulation of BMP signal strength may be used to reset the enterocyte "rheostat" of carbohydrate versus lipid uptake and to control the antimicrobial response through goblet cells.


Assuntos
Enterócitos , Células Caliciformes , Animais , Proteínas Morfogenéticas Ósseas/metabolismo , Diferenciação Celular , Enterócitos/metabolismo , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Camundongos
5.
Pediatr Surg Int ; 38(5): 755-759, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35235011

RESUMO

PURPOSE: Cell therapy is a promising approach to treat enteric neuropathies such as Hirschsprung disease (HD). Recent studies have reported that enteric neurons derived from stem cells (ENCCs) can be grafted into the HD colon. Thus, we investigated the migration and generation of enteric neurospheres from SOX10-VENUS+ mice after transplantation into control or Ednrb KO mice, which are a model of HD. METHODS: Single-cell suspensions were isolated from the fetal guts of SOX10-VENUS+ mice E13.5 and dissociated. These cells were cultured for 7 days under non-adherent conditions to generate neurospheres, which were co-cultured with dissociated control or SOX10-VENUS- Ednrb KO mouse gut on E13.5. 4 days later, these cells were fixed and the expression of the neuronal marker, Tuj1, was evaluated. RESULTS: Transplanted neurospheres had undergone abundant neuronal migration and differentiation of ENCCs in the control gut compared with the HD gut. The average length and intersections were significantly decreased in HD colon compared with controls (p < 0.05), and a similar pattern was observed in the HD small intestine (p < 0.05). CONCLUSIONS: We demonstrated that transplanted ENCCs did not differentiate properly in HD gut. These results highlight the importance of the neuronal environment in the recipient gut for enteric nervous system development.


Assuntos
Sistema Nervoso Entérico , Doença de Hirschsprung , Animais , Diferenciação Celular/fisiologia , Sistema Nervoso Entérico/metabolismo , Doença de Hirschsprung/cirurgia , Humanos , Intestino Delgado/metabolismo , Camundongos , Crista Neural/metabolismo
6.
J Proteome Res ; 21(4): 910-920, 2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35263542

RESUMO

Extracellular vesicles (EVs) mediate communication in physiological and pathological conditions. In the pathogenesis of type 2 diabetes, inter-organ communication plays an important role in its progress and metabolic surgery leads to its remission. Moreover, gut dysbiosis is emerging as a diabetogenic factor. However, it remains unclear how the gut senses metabolic alterations and whether this is transmitted to other tissues via EVs. Using a diet-induced prediabetic mouse model, we observed that protein packaging in gut-derived EVs (GDE), specifically the small intestine, is altered in prediabetes. Proteins related to lipid metabolism and to oxidative stress management were more abundant in prediabetic GDE compared to healthy controls. On the other hand, proteins related to glycolytic activity, as well as those responsible for the degradation of polyubiquitinated composites, were depleted in prediabetic GDE. Together, our findings show that protein packaging in GDE is markedly modified during prediabetes pathogenesis, thus suggesting that prediabetic alterations in the small intestine are translated into modified GDE proteomes, which are dispersed into the circulation where they can interact with and influence the metabolic status of other tissues. This study highlights the importance of the small intestine as a tissue that propagates prediabetic metabolic dysfunction throughout the body and the importance of GDE as the messengers. Data are available via ProteomeXchange with identifier PXD028338.


Assuntos
Diabetes Mellitus Tipo 2 , Vesículas Extracelulares , Estado Pré-Diabético , Animais , Diabetes Mellitus Tipo 2/metabolismo , Vesículas Extracelulares/metabolismo , Intestino Delgado/metabolismo , Camundongos , Estado Pré-Diabético/metabolismo , Proteoma/genética , Proteoma/metabolismo , Proteômica
7.
Eur J Pharm Biopharm ; 173: 92-102, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35227857

RESUMO

Gastrointestinal (GI) mucus is continuously secreted and lines the entire length of the GI tract. Essential for health, it keeps the noxious luminal content away from the epithelium. Our aim was to characterize the composition and structure of mucus throughout the various GI segments in dog. Mucus was collected from the stomach, small intestine (duodenum, jejunum, ileum), and large intestine (cecum, proximal and distal colon) from dogs. Composition was determined by multi-omics. Structural properties were investigated using cryoSEM and rheology. GI mucus contained 74-95% water and maintained a pH around 6.5. The proteome was similar across the different GI segments. The highest abundant secreted gel-forming mucin in the gastric mucus was mucin 5AC, whether mucin 2 had highest abundance in the intestinal mucus. Lipid and metabolite abundance was generally higher in the jejunal mucus than the colonic mucus. CryoSEM microscopy revealed smaller pore size in small intestinal mucus, which increased in the large intestine. All mucus samples showed shear-thinning behavior and characteristics of gel-like structure. In conclusion, the mucus is a highly viscous and hydrated material. These data provide an important baseline for future studies on human and canine intestinal diseases and the dog model in drug absorption.


Assuntos
Intestino Delgado , Muco , Animais , Colo/metabolismo , Cães , Trato Gastrointestinal/metabolismo , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Muco/metabolismo , Estômago
8.
J Anim Sci ; 100(4)2022 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-35323927

RESUMO

The impact of individual amino acids (AA) on gut hormone secretion and appetite regulation in pigs remains largely unknown. The aim of the present study was to determine the effect of the 20 proteinogenic AA on the release of the anorexigenic hormones cholecystokinin (CCK) and glucagon-like peptide 1 (GLP-1) in postweaning pigs. Six 25-d-old male piglets (Domestic Landrace × Large White; body weight = 6.94 ± 0.29 kg) were humanely killed for the collection of intestinal segments from the duodenum, jejunum, and ileum. Tissue samples from the three intestinal segments were used to determine which of the regions were more relevant for the analysis of gut peptides. Only the segments with the highest CCK and GLP-1 secretion and expression levels were evaluated with the 20 individual AA. Tissue segments were cut open, cleaned, and stripped of their muscle layer before identical circular samples were collected and incubated in 24-well plates for 1 h (37 °C, 5% v/v CO2). The culture broth consisted of a glucose-free KRB buffer containing no added AA (control) or with the addition of 10 mM of 1 of the 20 proteinogenic AA. Following incubation, tissues and supernatant were collected for gene expression and secretion analysis of CCK and GLP-1 levels. CCK secretion and mRNA expression were higher (P < 0.05) in duodenum when compared with proximal jejunum or ileum, whereas GLP-1/proglucagon levels were higher in ileum vs. duodenum (P < 0.05) and jejunum (P < 0.05, for GLP-1 only) in postweaning pigs. Based on these results, the effect of AA on CCK and GLP-1 secretion was studied in the duodenum and ileum, respectively. None of the AA tested stimulated both anorexigenic hormones. Of all the essential AA, Ile, Leu, Met, and Trp significantly (P < 0.05) stimulated GLP-1 from the ileum, while only Phe stimulated CCK from the duodenum. Of the nonessential AA, amide AA (Gln and Asn) caused the release of CCK, while Glu and Arg increased the release of GLP-1 from the ileum. Interpreting the results in the context of the digestion and absorption dynamics, non-bound AA are quickly absorbed and have their effect on gut peptide secretion limited to the proximal small intestine (i.e., duodenum), thus, mainly CCK. In contrast, protein-bound AA would only stimulate CCK release from the duodenum through feedback mechanisms (such as through GLP-1 secreted mainly in the ileum).


Understanding which dietary amino acids (AA) may impact the release of gut hormones involved in the modulation of feed intake, such as cholecystokinin (CCK) and glucagon-like peptide 1 (GLP-1), can help improve pig feed formulations. The series of studies presented assessed the effect of the 20 proteinogenic non-bound AA on the secretion of CCK and/or GLP-1 by duodenum, jejunum, and/or ileum samples from postweaning piglets. None of the AA tested stimulated the secretion of both CCK and GLP-1. Among the essential AA (EAA), Ile, Leu, Met, and Trp significantly stimulated GLP-1 from the ileum, while Phe stimulated CCK from the duodenum. Of the non-essential AA (NEAA), AA amides Gln and Asn caused the release of CCK, while Glu and Arg increased the release of GLP-1 from the ileum. The results suggest that both non-bound EAA and NEAA participate in appetite control via the release of gut peptides in pigs. Given that CCK was mainly released from duodenum samples (in the pre-enzymatic section of the small intestine), protein-bound AA could only influence CCK release through feedback mechanisms such as through the presence of GLP-1 receptors.


Assuntos
Colecistocinina , Peptídeo 1 Semelhante ao Glucagon , Aminoácidos/metabolismo , Animais , Duodeno/metabolismo , Intestino Delgado/metabolismo , Masculino , Suínos
9.
Biochim Biophys Acta Mol Cell Biol Lipids ; 1867(6): 159151, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35296424

RESUMO

Triacylglycerols are a major source of stored energy that are obtained either from the diet or can be synthesized to some extent by most tissues. Alterations in pathways of triacylglycerol metabolism can result in their excessive accumulation leading to obesity, insulin resistance, cardiovascular disease and nonalcoholic fatty liver disease. Most tissues in mammals synthesize triacylglycerols via the glycerol 3-phosphate pathway. However, in the small intestine the monoacylglycerol acyltransferase pathway is the predominant pathway for triacylglycerol biosynthesis where it participates in the absorption of dietary triacylglycerol. In this review, the enzymes that are part of both the glycerol 3-phosphate and monoacylglycerol acyltransferase pathways and their contributions to intestinal triacylglycerol metabolism are reviewed. The potential of some of the enzymes involved in triacylglycerol synthesis in the small intestine as possible therapeutic targets for treating metabolic disorders associated with elevated triacylglycerol is briefly discussed.


Assuntos
Intestinos , Metabolismo dos Lipídeos , Animais , Intestino Delgado/metabolismo , Mamíferos/metabolismo , Obesidade/metabolismo , Triglicerídeos/metabolismo
10.
PLoS One ; 17(3): e0264977, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35324937

RESUMO

In humans, celiac disease (CeD) is a T-cell-driven gluten-sensitive enteropathy (GSE) localized to the small bowel (duodenum). The presence of antibodies specific for gluten- and self-antigens are commonly used diagnostic biomarkers of CeD and are considered to play a role in GSE pathogenesis. Previously, we have described an apparent T-cell-mediated GSE in CD19-/- mice, which develop weak and abnormal B cell responses. Here, we expand on this observation and use a mouse model of complete B cell deficiency (JH-/- mice), to show that absence of a humoral immune response also promotes development of a GSE. Furthermore, 16S analysis of microbial communities in the small intestine demonstrates that a gluten-free diet suppresses the expansion of anaerobic bacteria in the small intestine and colonization of the small intestine by a specific pathobiont. Finally, we also observe that SI enteropathy in mice fed a gluten-rich diet is positively correlated with the abundance of several microbial peptidase genes, which supports that bacterial metabolism of gluten may be an important driver of GSE in our model. Collectively, results from our experiments indicate that JH-/- mice will be a useful resource to investigators seeking to empirically delineate the contribution of humoral immunity on GSE pathogenesis, and support the hypothesis that humoral immunity promotes tolerance to gluten.


Assuntos
Doença Celíaca , Animais , Dieta Livre de Glúten , Duodeno/metabolismo , Glutens/efeitos adversos , Intestino Delgado/metabolismo , Camundongos
11.
Cell Rep ; 38(13): 110560, 2022 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-35354041

RESUMO

It is not clear how the complex interactions between diet and intestinal immune cells protect the gut from infection. Neutral ceramidase (NcDase) plays a critical role in digesting dietary sphingolipids. We find that NcDase is an essential factor that controls intestinal immune cell dynamics. Mice lacking NcDase have reduced cluster of differentiation (CD) 8αß+ T cells and interferon (IFN)-γ+ T cells and increased macrophages in the intestine and fail to clear bacteria after Citrobacter rodentium infection. Mechanistically, cellular NcDase or extracellular vesicle (EV)-related NcDase generates sphingosine, which promotes macrophage-driven Th1 immunity. Loss of NcDase influences sphingosine-controlled glycolytic metabolism in macrophages, which regulates the bactericidal activity of macrophages. Importantly, administration of dietary sphingomyelin and genetic deletion or pharmacological inhibition of SphK1 can protect against C. rodentium infection. Our findings demonstrate that sphingosine profoundly alters macrophage glycolytic metabolism, leading to intestinal macrophage activation and T cell polarization, which prevent pathogen colonization of the gut.


Assuntos
Ceramidase Neutra , Esfingosina , Animais , Homeostase , Intestino Delgado/metabolismo , Macrófagos/metabolismo , Camundongos , Ceramidase Neutra/genética , Ceramidase Neutra/metabolismo , Esfingosina/metabolismo
12.
J Control Release ; 343: 584-599, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35149142

RESUMO

Micro- and nano-scale particulate formulations are widely investigated towards improving the oral bioavailability of both biologics and drugs with low solubility and/or low intestinal permeability. Particulate formulations harnessing physiological intestinal transport pathways have recently yielded remarkably high oral bioavailabilities, illustrating the need for better understanding the specific pathways underpinning particle small intestinal absorption and the relative role of intestinal cells. Mechanistic knowledge has been hampered by the well acknowledged limitations of current in vitro, in vivo and ex vivo models relevant to the human intestinal physiology and the lack of standardization in studies reporting absorption data. Here we review the relevant literature and critically discusses absorption pathways with a focus on the role of specific intestinal epithelial and immune cells. We conclude that while Microfold (M) cells are a valid target for oral vaccines, enterocytes play a greater role in the systemic bioavailability of orally administrated particulate formulations, particularly within the sub-micron size range. We also comment on less-reported mechanisms such as paracellular permeability of particles, persorption due to cell damage and uptake by migratory immune cells.


Assuntos
Absorção Intestinal , Intestino Delgado , Administração Oral , Disponibilidade Biológica , Transporte Biológico , Humanos , Intestino Delgado/metabolismo , Permeabilidade
13.
Cells ; 11(3)2022 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-35159145

RESUMO

Enteric glial cells (EGC) are known to regulate gastrointestinal functions; however, their role in Crohn's disease (CD) is elusive. Microscopic erosions over the ileal Peyer's patches are early signs of CD. The aim of this work was to assess the localization of EGC in the follicle and interfollicular region of the Peyer's patches and in the lamina propria and study the effects of EGC mediators on barrier function in CD patients and non-inflammatory bowel disease (non-IBD) controls. EGC markers, glial fibrillary acidic protein (GFAP), and S100 calcium-binding protein ß (S100ß) were quantified by immunofluorescence and Western blotting. Both markers showed significantly more EGC in the Peyer's patches and lamina propria of CD patients compared to the non-IBD controls. In CD patients there were significantly more EGC in Peyer's patches compared to lamina propria, while the opposite pattern was seen in controls. Barrier function studies using Ussing chambers showed increased paracellular permeability by EGC mediators in CD patients, whereas permeability decreased by the mediators in controls. We show the accumulation of EGC in Peyer's patches of CD patients. Moreover, EGC mediators induced barrier dysfunction in CD patients. Thus, EGC might have harmful impacts on ongoing inflammation and contribute to the pathophysiology of the disease.


Assuntos
Doença de Crohn , Doença de Crohn/metabolismo , Humanos , Intestino Delgado/metabolismo , Neuroglia , Permeabilidade , Nódulos Linfáticos Agregados
14.
Clin Pharmacol Ther ; 111(5): 1142-1154, 2022 05.
Artigo em Inglês | MEDLINE | ID: mdl-35158408

RESUMO

Mathematical models, such as physiologically-based pharmacokinetic models, are used to predict, for example, drug disposition and toxicity. However, populations differ in the abundance of proteins involved in these processes. To improve the building and refinement of such models, they must take into account these interindividual variabilities. In this study, we used global proteomics to characterize the protein composition of jejunum and liver from 37 donors with obesity enrolled in the COCKTAIL study. Liver protein levels from the 37 donors were further compared with those from donors without obesity. We quantified thousands of proteins and could present the expression of several drug-metabolizing enzymes, for the first time, in jejunum, many of which belong to the cytochrome P450 (CYP) (e.g., CYP2U1) and the amine oxidase (flavin-containing) (e.g., monoamine oxidase A (MAOA)) families. Although we show that many metabolizing enzymes had greater expression in liver, others had higher expression in jejunum (such as, MAOA and CES2), indicating the role of the small intestine in extrahepatic drug metabolism. We further show that proteins involved in drug disposition are not correlated in the two donor-matched tissues. These proteins also do not correlate with physiological factors such as body mass index, age, and inflammation status in either tissue. Furthermore, the majority of these proteins are not differently expressed in donors with or without obesity. Nonetheless, interindividual differences were considerable, with implications for personalized prediction models and systems pharmacology.


Assuntos
Sistema Enzimático do Citocromo P-450 , Jejuno , Sistema Enzimático do Citocromo P-450/metabolismo , Família 2 do Citocromo P450/metabolismo , Humanos , Intestino Delgado/metabolismo , Jejuno/metabolismo , Fígado/metabolismo , Obesidade/metabolismo
15.
PLoS One ; 17(2): e0264459, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35213654

RESUMO

Overt hepatic encephalopathy (HE) is one of the complications of liver cirrhosis (LC), which negatively affects the prognosis and quality of life of patients. Small intestinal bacterial overgrowth (SIBO) is significantly associated with LC and its complications, including HE. We investigated the relationship between SIBO and LC, and the difference between hydrogen-producing and methane-producing SIBO (H-SIBO and M-SIBO, respectively). This is a prospective cohort study of 107 cases. Breath measurements of hydrogen and methane concentrations were performed for the diagnosis of SIBO. The study cohort included 81 males with a median age of 70 (40-86) years, and SIBO was detected in 31 cases (29.0%). There were no significant differences between the SIBO positive and SIBO negative groups. Reclassification into H-SIBO (16 cases) and others (91 cases) was performed, and the Child-Pugh score was only derived in the multivariate logistic analysis (P = 0.028, odds ratio 1.39, 95% confidence interval 1.04-1.85). Furthermore, H-SIBO was significantly associated with covert HE in chi-square test (50.0% vs. 24.2%, P = 0.034). In addition, we evaluated the therapeutic response on SIBO of rifaximin in eight covert HE patients. 20% patients with M-SIBO and 67% patients with H-SIBO showed an improvement of the breath test. In conclusion, H-SIBO, but not M-SIBO, is significantly associated with liver function, and rifaximin might be more effective for covert HE with H-SIBO. Therefore, the diagnosis of SIBO, including the classification as H-SIBO and M-SIBO, might help to determine the choice of treatment for HE.


Assuntos
Microbioma Gastrointestinal/efeitos dos fármacos , Encefalopatia Hepática , Hidrogênio/metabolismo , Intestino Delgado , Fígado/metabolismo , Rifaximina/administração & dosagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Testes Respiratórios , Feminino , Encefalopatia Hepática/diagnóstico , Encefalopatia Hepática/tratamento farmacológico , Encefalopatia Hepática/metabolismo , Encefalopatia Hepática/microbiologia , Humanos , Intestino Delgado/metabolismo , Intestino Delgado/microbiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
16.
Nat Commun ; 13(1): 715, 2022 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-35132078

RESUMO

Organs are anatomically compartmentalised to cater for specialised functions. In the small intestine (SI), regionalisation enables sequential processing of food and nutrient absorption. While several studies indicate the critical importance of non-epithelial cells during development and homeostasis, the extent to which these cells contribute to regionalisation during morphogenesis remains unexplored. Here, we identify a mesenchymal-epithelial crosstalk that shapes the developing SI during late morphogenesis. We find that subepithelial mesenchymal cells are characterised by gradients of factors supporting Wnt signalling and stimulate epithelial growth in vitro. Such a gradient impacts epithelial gene expression and regional villus formation along the anterior-posterior axis of the SI. Notably, we further provide evidence that Wnt signalling directly regulates epithelial expression of Sonic Hedgehog (SHH), which, in turn, acts on mesenchymal cells to drive villi formation. Taken together our results uncover a mechanistic link between Wnt and Hedgehog signalling across different cellular compartments that is central for anterior-posterior regionalisation and correct formation of the SI.


Assuntos
Proteínas Hedgehog/metabolismo , Mucosa Intestinal/metabolismo , Intestino Delgado/embriologia , Células-Tronco Mesenquimais/metabolismo , Via de Sinalização Wnt/fisiologia , Animais , Linhagem da Célula , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Hedgehog/genética , Mucosa Intestinal/citologia , Mucosa Intestinal/embriologia , Intestino Delgado/citologia , Intestino Delgado/metabolismo , Células-Tronco Mesenquimais/citologia , Camundongos , Morfogênese , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/metabolismo , Via de Sinalização Wnt/genética
17.
Mol Med Rep ; 25(3)2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35088892

RESUMO

Compared with the available drugs for the treatment of fibrosis in other organs, the development of intestinal anti­fibrosis drugs is limited. Therefore, it is of practical significance to examine novel drugs to delay or block the development of intestinal fibrosis. The present study aimed to investigate the effect of atractylenolide III (ATL­III) on intestinal fibrosis. An MTT assay was used to detect the effect of ATL­III on the activity of IEC­6 cells. The migration and invasion of fibrotic cells stimulated with TGF­ß were determined via wound healing and Transwell assays. An immunofluorescence assay and western blotting were conducted to assess the expression levels of protein associated with epithelial­mesenchymal transition (EMT). The role of the AMP­activated protein kinase (AMPK) pathway was verified using compound C (an AMPK inhibitor) treatment. The results of the present study indicated that ATL­III had no effect on the cells at a dose of 1­20 µmol/l. Moreover, ATL­III can inhibit the invasion and migration of cells induced by TGF­ß1, as well as block the EMT process. It was found that ATL­III could also activate the AMPK pathway. Furthermore, compound C reduced the inhibitory effect of ATL­III on stimulated cells, which indicated that the AMPK pathway plays a role in the inhibition process. In conclusion, ATL­III may inhibit the EMT of IEC­6 cells stimulated with TGF­ß1 by activating the AMPK signaling pathway.


Assuntos
Proteínas Quinases Ativadas por AMP , Transição Epitelial-Mesenquimal , Proteínas Quinases Ativadas por AMP/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Células Epiteliais/metabolismo , Intestino Delgado/metabolismo , Lactonas , Sesquiterpenos , Transdução de Sinais
18.
J Clin Endocrinol Metab ; 107(5): e1969-e1975, 2022 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-34999838

RESUMO

CONTEXT: Small-intestinal neuroendocrine tumors (SI-NETs) have a modest but significantly higher prevalence and worse prognosis in male patients. OBJECTIVE: This work aims to increase understanding of this sexual dimorphism in SI-NETs. PATIENTS AND METHODS: Retrospectively, SI-NET patients treated in a single tertiary center were included and analyzed for disease characteristics. Estrogen receptor 1 (ESR1) and 2 (ESR2), progesterone receptor (PGR), and androgen receptor (AR) messenger RNA (mRNA) expression was assessed in primary tumors and healthy intestine. Estrogen receptor alpha (ERα) and AR protein expression were analyzed by immunohistochemistry in primary tumors and mesenteric metastases. RESULTS: Of the 559 patients, 47% were female. Mesenteric metastasis/fibrosis was more prevalent in men (71% / 46%) than women (58% / 37%; P = 0.001 and P = 0.027, respectively). In women, prevalence of mesenteric metastases increased gradually with age from 41.1% in women <50 years to 71.7% in women >70 years. Increased expression of ESR1 and AR mRNA was observed in primary tumors compared to healthy intestine (both P < 0.001). ERα staining was observed in tumor cells and stroma with a strong correlation between tumor cells of primary tumors and mesenteric metastases (rho = 0.831, P = 0.02), but not in stroma (rho = -0.037, P = 0.91). AR expression was only found in stroma. CONCLUSION: Sexual dimorphism in SI-NETs was most pronounced in mesenteric disease, and the risk of mesenteric metastasis in women increased around menopause. The combination of increased ERα and AR expression in the SI-NET microenvironment suggests a modulating role of sex steroids in the development of the characteristic SI-NET mesenteric metastasis and associated fibrosis.


Assuntos
Neoplasias Intestinais , Tumores Neuroendócrinos , Receptor alfa de Estrogênio/genética , Receptor alfa de Estrogênio/metabolismo , Feminino , Fibrose , Humanos , Neoplasias Intestinais/epidemiologia , Neoplasias Intestinais/genética , Neoplasias Intestinais/metabolismo , Intestino Delgado/metabolismo , Masculino , Pessoa de Meia-Idade , Tumores Neuroendócrinos/epidemiologia , Tumores Neuroendócrinos/genética , Tumores Neuroendócrinos/metabolismo , Prevalência , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Estudos Retrospectivos , Caracteres Sexuais , Microambiente Tumoral
19.
Biochem Biophys Res Commun ; 595: 7-13, 2022 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-35091109

RESUMO

The intestinal tract is an essential component of the body's immune system, and is extremely sensitive to exposure of ionizing radiation. While ionizing radiation can effectively induce multiple forms of cell death, whether it can also promote ferroptosis in intestinal cells and the possible interrelationship between ferroptosis and intestinal immune function has not been reported so far. Here, we found that radiation-induced major ultrastructural changes in mitochondria of small intestinal epithelial cells and the changes induced in iron content and MDA levels in the small intestine were consistent with that observed during cellular ferroptosis, thus suggesting occurrence of ferroptosis in radiation-induced intestinal damage. Moreover, radiation caused a substantial increase in the expression of ferroptosis-related factors such as LPCAT3 and ALOX15 mRNA, augmented the levels of immune-related factors INF-γ and TGF-ß mRNA, and decreased the levels of IL-17 mRNA thereby indicating that ionizing radiation induced ferroptosis and impairment of intestinal immune function. Liproxstatin-1 is a ferroptosis inhibitor that was found to ameliorate radiation-induced ferroptosis and promote the recovery from immune imbalances. These findings supported the role of ferroptosis in radiation-induced intestinal immune injury and provide novel strategies for protection against radiation injury through regulation of the ferroptosis pathway.


Assuntos
Ferroptose/fisiologia , Intestinos/patologia , Quinoxalinas/farmacologia , Lesões Experimentais por Radiação/prevenção & controle , Radiação Ionizante , Compostos de Espiro/farmacologia , 1-Acilglicerofosfocolina O-Aciltransferase/genética , 1-Acilglicerofosfocolina O-Aciltransferase/metabolismo , Animais , Araquidonato 12-Lipoxigenase/genética , Araquidonato 12-Lipoxigenase/metabolismo , Araquidonato 15-Lipoxigenase/genética , Araquidonato 15-Lipoxigenase/metabolismo , Ferroptose/efeitos dos fármacos , Ferroptose/efeitos da radiação , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Glutationa/metabolismo , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/metabolismo , Intestino Delgado/efeitos da radiação , Intestinos/efeitos dos fármacos , Intestinos/efeitos da radiação , Masculino , Malondialdeído/metabolismo , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Transmissão , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/efeitos da radiação , Mitocôndrias/ultraestrutura , Lesões Experimentais por Radiação/patologia , Lesões Experimentais por Radiação/fisiopatologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Superóxido Dismutase/metabolismo
20.
Biochem Biophys Res Commun ; 595: 14-21, 2022 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-35093635

RESUMO

Organoid cryopreservation method is one of key step in the organoid culture. We aimed to establish a simple and efficient cryopreservation method for mouse small intestinal organoids (MIOs) and colon organoids (MCOs) using various concentrations of cryoprotectant. Based on the theoretical simulation, we optimized the dimethyl sulfoxide (DMSO) concentration by pretreating the organoids with 5, 7.5, and 10% DMSO for 30 min at 4 °C to allow penetration into the organoids and evaluated their viability, proliferation, and function after cryopreservation. Gene expression in the MIOs and staining of lineage markers were examined real-time PCR. The organoids in the DMSO-treated groups as well as the control, expressed ChrgA, Ecad, Muc2, Lyz, villin, and Lgr5, and there are no significant. A forskolin-induced swelling assay for MIOs was performed to confirm normal cystic fibrosis transmembrane conductance regulator (CFTR) activity. Similar forskolin-induced swelling was observed in the DMSO-treated groups and the control. In addition, MCOs were transplanted into mouse colon for confirmation of regeneration therapy efficacy. Thawing organoids were cultured for two and four sequential passages after cryopreservation with 5% DMSO to confirm any changes in the gene expression of lineage markers after subculture. We developed a simple and efficient organoid freezing method using 5% DMSO with low potential toxicity and validated our findings with theoretical simulation.


Assuntos
Colo/metabolismo , Criopreservação/métodos , Intestino Delgado/metabolismo , Organoides/metabolismo , Medicina Regenerativa/métodos , Animais , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Crioprotetores/metabolismo , Crioprotetores/farmacologia , Dimetil Sulfóxido/metabolismo , Dimetil Sulfóxido/farmacologia , Expressão Gênica/efeitos dos fármacos , Camundongos , Organoides/citologia , Organoides/efeitos dos fármacos , Fatores de Tempo
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