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@#Abstract: To explore the mechanism of the intestinal microecology regulation by polysaccharide prebiotics, ELISA, histopathologic analysis, immunohistochemical analysis, 16S rRNA high-throughput sequencing, and gas chromatography-mass spectrometry were applied to investigate the effects of fermented polysaccharides on changes in the intestinal microbiota and short-chain fatty acids (SCFAs) in mice with dextran sulfate sodium (DSS)-induced colitis model and their relationship with the level of intestinal inflammation and barrier protein expression. It was found that fermented Lycium barbarum polysaccharides (FLBP) significantly reduced intestinal inflammation level, improved colonic tissue structure, up-regulated the expression of tight junction proteins Claudin-1 and ZO-1, and significantly increased the content of intestinal SCFAs in mice. Gut bacteria analyses showed that FLBP enriched intestinal Dubosiella and Akkermansia in mice and decreased the abundance of Turicibacter, Faecalibaculum, and Escherichia-Shigella. Results showed that remodeled Dubosiella activated by FLBP played a dominant role in ameliorating colitis by significantly increasing SCFAs content, improving intestinal barrier and reducing intestinal inflammation. The study aimed to provide a safer and better option for the amelioration of colitis and to provide a theoretical basis for the development of functional foods with FLBP.
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Objective:To discuss the improvement effect of curcumin combined with fecal bacteria transplantation on the mice with dextran sulfate sodium(DSS)-induced ulcerative colitis(UC),and to clarify the related mechanism.Methods:Fifty mice were randomly divided into control,model,curcumin,fecal bacteria transplantation,and combination groups.Except for the mice in control group(given distilled water),the mice in the other groups were given distilled water containing 2%DSS to establish the UC models.The mice in curcumin group were gavaged with 0.4 mL of 60 mg·kg-1 curcumin solution once per day for 10 d;the mice in fecal bacteria transplantation group underwent enema with 0.2 mL of fecal bacteria suspension once per day for 10 d;the mice in combination group received the enema of 0.2 mL fecal bacteria suspension and gavaged with 0.4 mL of 60 mg·kg-1 curcumin solution.At the end of the experiment,the disease activity index(DAI)and colon macroscopic damage index(CDMI)of the mice in various groups were calculated;the morphology of colon tissue of the mice in various groups was detected by HE staining;the levels of interleukin(IL)-1β,tumor necrosis factor-α(TNF-α),IL-4,and IL-10 in colon tissue of the mice in various groups were detected by enzyme-linked immunosorbent assay(ELISA)method;the expression levels of occludin and zonula occludens-1(ZO-1)mRNA and proteins in colon tissue of the mice in various groups were detected by real-time fluorescence quantitative(RT-qPCR)and Western blotting methods.Results:The intestinal mucosal epithelial structure of the mice in control group was intact and continuous with regular glandular arrangement and without inflammatory cell infiltration or ulceration;the intestinal mucosal epithelial structure of the mice in model group exhibited loss of colonic mucosal epithelium,disordered glandular arrangement,reduced goblet cells,congestion and edema in mucosal and submucosal layers,and extensive infiltration of inflammatory cells with widespread small ulcers;the intestinal mucosal epithelial structure of the mice in curcumin,fecal bacteria transplantation,and combination groups exhibited relatively intact colonic mucosal epithelial structures with reduced inflammatory cell infiltration and ameliorated mucosal and submucosal congestion and edema.Compared with control group,the DAI and CDMI of the mice in model group were increased(P<0.05),the levels of IL-1β and TNF-α were increased(P<0.05),the levels of IL-4 and IL-10 were decreased(P<0.05),and the expression levels of occludin and ZO-1 mRNA and proteins were decreased(P<0.05);compared with model group,the DAI and CDMI of the mice in curcumin,fecal bacteria transplantation,and combination groups were decreased(P<0.05),the levels of IL-1β and TNF-α were decreased(P<0.05),the levels of IL-4 and IL-10 were increased(P<0.05),and the expression levels of occludin and ZO-1 mRNA and proteins were increased(P<0.05).Compared with curcumin group and fecal bacteria transplantation group,the DAI and CDMI of the mice in combination group were decreased(P<0.05),the levels of IL-1β and TNF-α were decreased(P<0.05),the levels of IL-4 and IL-10 were increased(P<0.05),and the expression levels of occludin and ZO-1 mRNA and proteins were increased(P<0.05).Conclusion:Curcumin combined with fecal bacteria transplantation can ameliorate the pathological damage in colonic tissue of the UC mice,inhibit the secretion of inflammatory factors,and promote the repaiment of intestin mucosa.
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Objective To induce an NLRP3-/- mouse model of ulcerative colitis(UC)using different concentrations of dextran sulfate sodium(DSS)and different administration times,and to analyze and evaluate the advantages and disadvantages of the preparations to provide a more suitable animal model for the study of UC pathogenesis in humans and the development of therapeutic drugs.Methods Forty-eight male NLRP3-/- specific-pathogen-free mice were divided randomly into blank,2.5％7 d,3％7 d,and 3％5 d groups(n=12 mice per group).UC mouse models were induced using combinations of different concentrations and administration times of DSS.Body weight,DAI(disease activity index)score,hematoxylin and eosin(HE)staining,colon length,and related indicators(interleukin IL-6,tumor necrosis factor(TNF)-α,and tight junction protein(ZO-1))were observed and evaluated.Results(1)UC membrane type was induced in each group with different concentrations and administration times.(2)Mouse body weight decreased,the fecal occult blood became more positive,the DAI score increased,and more mice died with increasing DSS concentration and administration time.(3)Longer administration time and higher concentration of DSS were also associated with more severe damage to the intestinal mucosa,as shown by HE staining.(4)Immunohistochemistry showed that the inflammatory factors TNF-α and IL-6 were increased in the model group compared with the blank control group,while expression of ZO-1 was decreased compared with the blank group.Conclusions(1)Administration of 2.5％or 3％DSS for 7 days or 3％DSS for 5 days can induce UC in NLRP3-/- mice.(2)The combination of DAI score,HE staining,the detection of related indicators,and mouse survival rate indicated that NLRP3-/- mice treated with 3％DSS for 5 days produced the most suitable UC model to study the clinical manifestations and drug treatment of UC.
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Objective @#To compare acute ulcerative colitis (UC) model induced by dextran sulfate sodium ( DSS) at 1. 5% and 3. 0% concentrations and obtain a cost-effective modeling scheme with low mortality．@*Methods@#C57BL /6J mice were randomly divided into control group，1. 5% and 3. 0% DSS group．The mice in control group drank sterilized water and the mice in each group of DSS drank the corresponding concentration of DSS solution，all mice were treated for 7 days．And all mice drank sterilized water on day 8 to 14．The blood stool of mice was ob- served and the disease activity index，the percentage of body weight loss and the percent of survival of mice were calculated on day 1 to 14．Mice were sacrificed on day 8．The spleen index was calculated and the length of colon was measured ; HE staining was used to observe the histopathological changes of colon ; The expressions of tight junc- tion protein Claudin-1 and Occludin in colon tissue were detected by immunohistochemistry ; Immunofluorescence detected the ratio of macrophage in colon tissue ; Levels of tumor necrosis factor-α (TNF-α) in serum and colon tis- sue homogenates were detected by ELISA. @*Results @#Compared with the control group，both mice of 1. 5% DSS group and mice of 3. 0% DSS group showed similar clinical and pathological features to human UC ; The colon lengths were both shortened (P＜0. 01) ; The expressions of Claudin-1 and Occludin in colon tissue both decreased (P＜0. 01) ; Spleen index increased (P＜0. 01) ; The proportion of macrophages in colon tissue increased (P＜0. 01) ; Levels of TNF-α in serum and colon tissue homogenates both increased (P ＜0. 01 ) ．The percent of survival of 1. 5% DSS mice within 14 days was higher than that of 3. 0% DSS mice．@*Conclusion @#The acute UC model can be established efficiently and economically by giving 1. 5% concentration of DSS solution in mice for 7 days.
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AIM: To evaluate the clinical effect of Iron Dextran Dispersible Tablets on patients with chronic heart failure who reduced ejection fraction after 24 weeks. METHODS: From January 2020 to June 2022, forty-five patients with heart failure complicated with iron deficiency and reduced ejection fraction were selected as the research objects. According to the random number table, they were randomly divided into control group and observation group.The control group was given routine anti-heart failure treatment such as Sacubitril Calsartan sodium tablets, while the observation group was given iron dextran dispersible tablets 50 mg three times a day on the basis of the anti-heart failure treatment of the control group for 8 weeks. The 6-minute walking distance, Hemoglobin, Serum Ferritin, N-terminal B-type natriuretic peptide precursor, Left Ventricular Ejection Fraction, Left Ventricular end Diastolic Diameter and 12-item Kansas City Cardiomyopathy Questionnaire (KCCQ-12) overall summary score and clinical summary score were compared between the two groups. RESULTS: There was no significant difference in baseline data between the two groups (P > 0.05). After treatment, the 6-minute walking distance in the observation group was longer than that in the control group, while the serum ferritin level in the observation group was higher than that in the control group. The N-terminal pro-B-type natriuretic peptide level in the two groups was lower than that before treatment, and the left ventricular end diastolic diameter was shorter than that before treatment, and the left ventricular ejection fraction, clinical comprehensive score and symptom score were higher than that before treatment. The difference was statistically significant (P 0.05). CONCLUSION: Iron Dextran Dispersible Tablets can improve the exercise endurance and quality of life of patients with chronic heart failure who reduced ejection fraction after 24 weeks.
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ObjectiveTo explore the protective effect of Xielitang on ulcerative colitis （UC） mice induced by dextran sodium sulfate （DSS） and its possible mechanism. MethodSixty C57BL/6 mice were randomly divided into normal group， model group， sulfasalazine group and and low-， medium-， and high-dose Xielitang groups. Free drinking DSS solution to build the chronic UC model mice. Except for normal group， other groups were given 1.5% DSS for 3 cycles of drinking （days 1-7， days 22-28 and days 43-49） and distilled water for the rest of the time （days 8-21， days 29-42 and days 50-63）. After the first cycle， corresponding drugs were given for 42 days. The changes of general condition， body weight and disease activity index （DAI） score of mice were daily recorded during the experiment. At the end of the treatment， serum and colon tissue samples were collected， colon length was measured， intestinal weight index and colonic mucosal injury （CMDI） score were calculated. The pathological status of colon tissue was observed by hematoxylin-eosin （HE） staining. The levels of interleukin-6 （IL-6）， interleukin-10 （IL-10） and tumour necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay （ELISA）. The gene and protein expressions of Toll like receptor 4 （TLR4）， nuclear transcription factor-κB （NF-κB） and hypoxia inducible factor-1α （HIF-1α） in colon tissue was detected by Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultCompared with the normal group， the body weight， colon length and IL-10 content in the model group were significantly decreased （P<0.01）， DAI score， intestinal weight index， CMDI score， IL-6 and TNF-α contents， and mRNA and protein expression levels of TLR4， NF-κB and HIF-1α in the model group were significantly increased （P<0.01）. Moreover， the structure of colonic mucosa was destroyed and inflammatory cells infiltrated in the model group. Compared with model group， body weight， colon length and IL-10 content in each dose group of Xielitang were significantly increased （P<0.05， P<0.01）， DAI score， intestinal weight index and CMDI score， IL-6 and TNF-α contents， mRNA and protein expression levels of TLR4， NF-κB and HIF-1α were notably decreased （P<0.05， P<0.01）. The pathological injury of colon was obviously alleviated. ConclusionXielitang can significantly improve the inflammatory response of UC mice induced by DSS， and its mechanism may be related to the regulation of TLR4/NF-κB/HIF-1α signaling pathway.
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Objective:To investigate the effects of piperine on experimental colon carcinogenesis induced by 1, 2-dimethylhydrazine (DMH)/sodium dextran sulfate (DSS) and its mechanism.Methods:36 mice were divided into control group, model group and piperine group, 12 mice in each group. The control group was given normal saline by gavage; The model group and piperine group were given 3.6 mg/(kg·d) of normal saline and piperine respectively after establishing the experimental colon cancer model induced by DMH/DSS. Tumor load and volume were observed. Hematoxylin and eosin (HE) staining was used to observe the histological change of colon in mice. RAS/PI3K/AKT related pathway protein expression was detected by Western blot.Results:The body weight gain, protein expression levels of cleaved poly-ADP ribose polymerase (PARP), cleaved caspase-3 in model group were significantly lower than those in the control group (all P<0.05). The protein expression levels of Bcl-2, Bax, pan-Ras, p-MEK, p-ERK, PI3K, p-AKT, NF-κBP65, c-Myc and cyclin D1 in model group were significantly higher than those in the control group (all P<0.05). The body weight gain, protein expression levels of cleaved PARP and cleaved caspase-3 in piperine group were significantly higher than those in model group (all P<0.05). The protein expression levels of Bcl-2, Bax, pan-Ras, p-MEK, p-ERK, PI3K, p-AKT, NF-κBP65, c-Myc and cyclin D1 in piperine group were significantly lower than those in model group (all P<0.05). Conclusions:Piperine can inhibit the occurrence of experimental colon cancer induced by DMH/DSS, which may involve multiple components of RAS/PI3K/AKT signal axis.
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【Objective】 To establish the optimal treatment model of Bletilla striata polysaccharide （BSP） on dextran sodium sulfate （DSS）-induced ulcerative colitis （UC） in mice. 【Methods】 We randomly divided 48 male C57BL/6 mice into normal control group， DSS model group （25 g/L DSS）， BSP low-， medium- and high-dose groups （25 g/L DSS + 95， 190， 380 mg/kg BSP）， and salazosulfapyridine （SASP） （25 g/L DSS + 320 mg/kg SASP， positive control） group. Mice in the normal control group drank distilled water freely， while the other groups were given 25 g/L DSS solution to drink freely for 7 days. From the second day， the low-， medium- and high-dose BSP groups and SASP （positive control）group were administered by gavage according to body mass. The normal control group and DSS model group were given the same amount of normal saline once a day for 7 consecutive days. The mice’s blood pressure was recorded every day. Mental state， body mass， stool characteristics and bloody stool were used to calculate the mice’s disease activity index （DAI）. The mice were killed on the 9th day， and their colonic tissues were taken for hematoxylin eosin （HE） staining and histopathological scoring. The expression of tight junction protein Claudin-1 in colonic tissues was detected by Western blotting. 【Results】 Compared with the normal control group， the DSS model group had obvious clinical manifestations， histopathological changes and reduced body weight， increased histopathological score and DAI score （P<0.05）， and decreased expression of tight junction protein Claudin-1 in colon tissue （P<0.05）. Compared with those in DSS model group， the clinical manifestations of UC and colonic mucosal injury in low-， medium- and high-dose BSP groups were improved in varying degrees. The high-dose （380 mg/kg） BSP group had the best effect. The degree of body weight reduction， histopathological score and DAI score in this group were significantly lower than those in DSS model group （P<0.05）， whereas the expression of Claudin-1 increased significantly （P<0.05）. 【Conclusion】 When BSP was administered by gavage at 380 mg/kg， the therapeutic effect on UC mice induced by 25 g/L DSS was the best. This model can be used as an effective one for further studies on Striata Bletilla polysaccharide in UC mice.
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n-3 polyunsaturated fatty acids (PUFAs) play an active role in controlling the progression of ulcerative colitis (UC), but its mechanism is not very clear. In this study, we compared the effects of fish oil (the main component is n-3 PUFAs) in the mouse model with acute and chronic colitis induced by dextran sulfate sodium (DSS). Male C57BL/6J mice were randomly divided into six groups, and each group had ten mice. The alleviating effect of fish oil on chronic colitis was significantly better than acute colitis as indicated by the following analysis: the weight loss of mice (P < 0. 05), decreased disease activity index (DAI) score (P<0. 05), colonic edema, colon length index and histopathological score (P < 0. 05), and serum pro-inflammatory factor levels like IL-1β, TNF-α, and IL-6 (P < 0. 01). Moreover, fish oil promoted the level of serum anti-inflammatory factor IL-10 (P<0. 05). The treatment of fish oil increased the n-3 PUFA concentration in the intestinal epithelium of mice (P < 0. 01), especially EPA (P<0. 05). 16S rRNA sequencing of feces revealed that fish oil significantly increased the relative abundance of butyrate-producing flora (Clostridiales) and probiotics (Bifidobacteriales) in the feces of the maintained remission model group, reduce the proportion of aerobic, parthenogenic anaerobic and pathogenic, and improved the disorder of glycan biosynthesis and metabolism and oxidative phosphorylation (P<0. 05). Compared with the induced remission fish oil group, fish oil treatment led to an elevated expression of mechanical barrier and energy metabolism pathway proteins in the maintained remission fish oil group. Our results showed that fish oil exerted a more potent inhibitory effect in the remission mice model, which may be related to effectively strengthening the mechanical barrier, improving the composition and function of intestinal microbiota and concentration of butyric acids and improving dysbiosis of host-microbial interaction.
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OBJECTIVES@#Ulcerative colitis (UC) is a chronic, relapsing inflammation of the colon. Impaired epithelial repair is an important biological features of UC. Accelerating intestinal epithelial repair to achieve endoscopic mucosal healing has become a key goal in UC. Yes-associated protein (YAP) is a key transcriptional coactivator that regulates organ size, tissue growth and tumorigenesis. Growing studies have focused on the role of YAP in intestinal epithelial regeneration. This study explore the molecular mechanism for the role YAP in modulating colonic epithelial proliferation, repair, and the development of colitis associated cancer.@*METHODS@#We constructed the acute colitis mouse model through successive 5 days of 3% dextran sulfate sodium salt (DSS) induction. Then YAP-overexpressed mouse model was constructed by intraperitoneal injection the YAP overexpressed and negative control lentivirus into DSS mice. On the 5th day of DSS induction and the 5th day of normal drinking water after removing DSS (5+5 d), the mice were killed by spinal dislocation. The colon was taken to measure the length, and the bowel 1-2 cm near the anal canal was selected for immunohistochemical and Western blotting. We used YAP over-expressed colonic epithelial cells and small interfering signal transducer and activator of transcription 3 (STAT3) RNA to probe the regulation of YAP on STAT3, using cell counting kit-8 and scratch assays to explore the role of YAP on colonic epithelial cell proliferation. Finally, we conducted co-immunoprecipitation to test the relationship between YAP and STAT3.@*RESULTS@#After DSS treatment, the expression of YAP was dramatically diminished in crypts. Compared with the empty control mice, overexpression of YAP drastically accelerated epithelial regeneration after DSS induced colitis, presenting with more intact of structural integrity in intestinal epithelium and a reduction in the number of inflammatory cells in the mucosa. Further Western blotting, functional experiment and co-immunoprecipitation analyses showed that the expression of YAP in nucleus was significantly increased by 2 h post DSS cessation, accompanied with up-regulated total protein levels of STAT3 and phosphorylated-STAT3 (p-STAT3). Overexpression of YAP enhanced the expression of STAT3, p-STAT3, and their transcriptional targets including c-Myc and Cyclin D1. In addition, it promoted the proliferation and the "wound healing" of colonic cells. However, these effects were reversed when silencing STAT3 on YAP-overexpressed FHC cells. Moreover, protein immunoprecipitation indicated that YAP could directly interact with STAT3 in the nucleus, up-regulatvng the expressvon of STAT3. Finally, during the process of CAC, overexpression of YAP mutant caused the down-regulated expression of STAT3 and inhibited the development and progress of CAC.@*CONCLUSIONS@#YAP activates STAT3 signaling in regulation of epithelial cell proliferation and promotes mucosal regeneration after DSS induced colitis, which may serve as a potential therapeutic target in UC. However, persistent and excessive YAP activation may promote CAC development.
الموضوعات
Animals , Mice , Cell Proliferation , Colitis/drug therapy , Colon/metabolism , Dextran Sulfate/adverse effects , Disease Models, Animal , Intestinal Mucosa , Mice, Inbred C57BL , Neoplasm Recurrence, Local/metabolism , STAT3 Transcription Factor/metabolism , YAP-Signaling Proteins/metabolismالملخص
Objective To investigate the role of TRIM65 on DSS induced colitis and the underlying molecular mechanisms. Methods Trim65+/+ and Trim65-/- mice were administered with 3% (w/v) DSS in their drinking water for 5 consecutive days and then were switched to sterile water for 2 days. DSS treated mice were monitored daily for the clinical symptoms (bodyweight, stool consistency and rectal bleeding score). Mice were sacrificed on day 7 to measure colon length. Colon homogenates were collected to measure MPO activity and detect cleaved caspase-1 and mature IL-1β by Enzyme linked immunosorbent assay (ELISA) and Western blot. Trim65-/- mice were intraperitoneally injected with NLRP3 inflammasome inhibitor MCC950, and were given the above treatment to determine the effect of MCC950 on colitis in Trim65-/- mice. Results The results showed that deletion of Trim65 significantly enhanced weight loss and colon shortening in DSS mice, increased disease activity index and histopathological score, induced the activity of MPO, and promoted the F4/80+ immune cell infiltration, the activation of caspase-1 and the secretion of mature IL-1 in the colon of DSS mice. The NLRP3 inflammasome inhibitor MCC950 alleviated DSS induced colitis symptoms and inflammation levels in trim65 deficient mice. Conclusion TRIM65 plays an anti-inflammatory role in DSS induced colitis mice by inhibiting the activation of NLRP3 inflammasome.
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Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.
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Aim To investigate the influence of different concentrations of dextran sulfate sodium (DSS) and administration approaches on the establishment of mouse model of acute ulcerative colitis(UC). Methods Mice were randomly divided into the normal control group, 3% DSS free drinking group, 4% DSS free drinking group, 30% DSS intragastric administration group and 40% DSS intragastric administration group. The amount and average dosage of intaken DSS by mice in free drinking group were measured and the intragastric administration group were given the same dosage. The survival rate, disease activity index (DAI), the lengths and pathological changes of colon were observed, and the coefficients of variation of each indicator described above were compared among the groups. Results Except for the normal control group, other groups developed experimental UC. Among these four approaches, 3% DSS solution free drinking showed appropriate incidence, higher animal survival rate and operability and lower cost, while there was no significant difference in the coefficients of variation of DAI between 3% DSS solution free drinking group and the others. Conclusion 3% DSS solution free drinking has more advantages in the establishment of a murine model with acute UC than other approaches.
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Ulcerative colitis is a most common form of inflammatory bowel disease (IBD) which mainly affect colon. The treatment of ulcerative colitis depends upon severity of the diseases. The aim of the present study was to determine the effect of hydroalcoholic extract of dried fruits of Helicteres isora in dextran sulfate sodium induced ulcerative colitis in experimental wistar rats. In this study wistar rats of either sex were divided into five experimental groups, where control group recived only distilled water. Group 2 was negative control group which received 4% dextran sulfate sodium (DSS) from drinking water between 15th to 21st day. Group 3 received low dose of hydroalcholic extract of Helicteres isora (HI) at a dose 100mg/kg orally along with 4% DSS from drinking water between from drinking water between 15th to 21st day. Group 4 received high dose of hydroalcholic extract of Helicteres isora (HI) at a dose 200mg/kg orally along with 4% DSS from drinking water between from drinking water between 15th to 21st day. In group 5 sulfasalazine was used as a standard drug at a dose 100mg/kg orally along with 4% DSS from drinking water between from drinking water between 15th to 21st day. Twenty four hours after treatment animals were sacrificed and further macroscopical, biochemical, histopathological evaluation was done and all the results were compare with control at p<0.05 significant value.
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Background: recently, it has been proved that copolymers with dextran cores and grafted polyacrylamide are effective in photodynamic and chemotherapy. However, further research is needed to define correct dosage and to assess the risks. Thus, animal studies are becoming more relevant to determine the effect of the treatment of such drug nano-systems on female reproductive function in particular.Methods: a technique for estimation of pre- and post-implantation death rates, in vitro meotic maturation of oocytes, double fluorescent vital assay and statistical analysis were used. The effects of a one-time treatment of different doses of dextran-polyacrylamide matrices and silver (Ag)-nanoparticles-dextran-polyacrylamide (AgNPs-D-PAA) on reproductive function, namely on 1) the number of oocytes isolated from one ovary and the meiotic maturation of such oocytes in vitro; 2) the indicators of cell viability of the cells of follicular environment of oocytes (FEO) and the cells of inguinal lymph nodes (ILN); 3) the pre- and post-implantation mortality rates and the number of live newborns (pups) were investigated in female mice.Results: no significant changes in the number of oocytes isolated from one ovary and meiotic maturation of such ovarian oocytes in vitro, the number of living cells of follicular environment of oocytes and the number of such cells with morphological signs of apoptosis and necrosis, pre- and post-implantation mortality rates of embryos and the number of live newborns (pups) have been established under conditions of one-time treatment with dextran-polyacrylamide at doses of 0.39 mg/kg and 3.90 mg/kg and Ag-nanoparticles-dextran-polyacrylamide at doses of 0.20 mg/kg and 2.00 mg/kg.Conclusions: branched polymer systems (dextran-polyacrylamide (D-PAA) polymer matrices) are promising materials for use in next-generation medicine.
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@#The conventional equilibrium dialysis and ultrafiltration methods cannot be used to determine the protein binding of some peptides because of their non-specific adsorption on the semipermeable membrane or poor stability in the plasma. The method of dextran-coated charcoal adsorption combined with LC-MS/MS were used. Based on the kinetic principle of initial rate of candidate drugs absorbed to dextran-coated charcoal, seven phosphorylated peptides with the same amino acid sequence and different configurations in rat plasma were selected as the study model using; the protein binding in rat plasma were determined; the amino acid distribution rules affecting the changes in protein binding rates of peptide candidate drugs were summarized. The results suggest that the dextran charcoal adsorption method, as a supplementary method for the determination of plasma protein binding, is suitable for peptides or organic drug candidates that cannot be determined by traditional techniques.
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Objective: To investigate the role of CD8+ T cells in the pathogenesis of acute murine colitis induced by dextran sulfate sodium (DSS). Methods: Wild type and CD8 knock-out (CD8-/-) mice with C57BL/6 background were given DSS with concentration of 2% (m/V). The body weight, colon length, pathological changes and disease activity of colitis were observed dynamically. The total RNA was extracted from the distal colon of mice after induction for 10 d. The mRNA expression of inflammatory cytokines Il1b, Il6, Il17a, Ifng, Tnf, Il10 and Tgfb1 were detected by real-time quantitative PCR. Colon tissue sections were stained with hematoxylin-eosin (H-E) and the changes of intestinal histopathology were evaluated, and the infiltration of CD8+ T cells in colon tissue was observed by immunofluorescence staining. The survival rate of mice was observed with 3% and 4% (m/V) DSS solution-induced colitis models. Results: After CD8-/- mice being induced by 2% DSS, the body weight decreased slowly and showed an increasing trend on the 9th day, while the pathological changes of colon tissues of CD8-/- mice were slight. The expression levels of Il1b, Il6, Il17a, Ifng and Tnf mRNA were lower than those of wild-type mice (P<0.05). The number of CD8+ T cells in colonic lamina propria of wild-type mice with 2% DSS induction was higher than that of wild-type mice without DSS treatment (P=0.001). The survival rates of wild-type mice induced by 3% and 4% DSS were 37.5% and 0, and the survival rates of CD8-/- mice were 66.7% and 100%, while the survival rates of CD8-/- mice receiving 3% and 4% DSS were higher than those of wild-type mice (P=0.025, P=0.001). Conclusion: CD8+ T cells can promote the development of murine acute DSS-induced colitis.
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Objective: To explore the factors affecting the nasal entry of the pharmaceutical preparations into the brain based on the established model of the "nose-brain" pathway in vitro. Methods: Calu-3 cells and OECs cells were co-cultured to construct a "nasal-brain" pathway cell model group. Taking fluorescein isothiocyanate dextran (FD) and fluorescent silver nanoparticles (AgNPs) as model drugs, the effects of drug molecular weight (Mw) factors and preparation particle size factors on the drug transnasal transport into the brain were explored. Results: The apparent permeability coefficient (Papp) of transcellular monolayer transport of FD decreased with the increase of molecular weight. The uptake of fluorescein isothiocyanate dextran with different molecular weights by OECs tended to be saturated after 90 min. As the molecular weight of FD increased, the uptake of OECs decreased significantly during the same uptake time. The apparent permeability coefficient of fluorescent AgNPs with different particle sizes in the "nose-brain" multi-channel cell model group of calu-3 monolayer decreased with the increase of the particle size of the nanoparticles. When the particle size was less than 40 nm, its transport characteristics in Calu-3 were shown as medium absorption (1 × 10-6 < Papp < 10 × 10-6), and when the particle size of nanoparticles was more than 60 nm, its transport characteristics were shown as difficult to absorb (Papp < 1 × 10-6). The uptake of OECs of fluorescent AgNPs with different particle sizes tended to be saturated at 60 min, and with the increase of the particle size of fluorescent AgNPs, the uptake of OECs at the same uptake time showed a significant decline. Conclusions The Mw of the drug and the particle size of the nano-formulation have an important influence on the nasal transport of the drug into the brain. Drugs with a molecular weight of < 4 000 and nano particles with a particle size of less than 40 nm have better transport and uptake characteristics.
الملخص
Objective: To investigate the effect of oligosaccharide grape seed proanthocyanins (GSPE) on dextran sulphate sodium salt (DSS)-induced ulcerative colitis (UC) in mice and its mechanisms. Methods: SPF-class C57 mice were randomly divided into normal group, model group, positive group (sulfasalazine group), and GSPE groups (125, 250, 500 mg/kg). The normal group was given pure water, and the other groups were free to drink 3% DSS aqueous solution for 7 d to induce the model of UC in mice. The changes of body weight, hematochezia and stool type were recorded every day. After seven days of treatment, blood, colons and spleens were collected, and the length of the colon and the weight of the spleen were recorded. HE staining was used to evaluate the pathological changes of colonic mucosa in mice. The expression of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor TNF-α in serum and colon tissues and the levels of NO, MDA, and SOD were detected by ELISA. The changes of HO-1, NF-κB, and Nrf2 in colonic epithelial cells were analyzed by immunohistochemistry. Results: Compared with the model group, the body weight of mice in GSPE groups (250, 500 mg/kg) decreased relatively slowly, and the symptoms of diarrhea and hematochezia were improved significantly. The content of IL-1β, IL-6, TNF-α, NO, and MDA in serum and colon tissues was much lower in the administration groups than those in the model group, while the content of SOD was significantly higher (P < 0.01). The pathological tissue analysis showed that the pathological damage of colonic mucosa in the high dose group of GSPE was obviously decreased. Immunohistochemical analysis showed that GSPE groups significantly decreased the expression of NF-κB and increased the expression of Nrf2 and HO-1 (P < 0.01). Conclusion: GSPE could effectively improve the symptoms of UC induced by DSS, and regulate the expression of oxidative stress-related proteins Nrf2, HO-1 and inflammatory pathway protein NF-κB, and then affect the changes of oxidative stress indicators SOD, MDA and inflammatory factors. Therefore, GSPE plays an important role in the treatment and prevention of UC.
الملخص
BACKGROUND: Crocin has anti-inflammatory and anti-oxidative stress effects. The therapeutic effects of crocin on ulcerative colitis and related mechanisms are still unclear. OBJECTIVE: To explore the protective effect of crocin in ulcerative colitis rats and its related mechanism. METHODS: Thirty Sprague-Dawley rats were randomly divided into five groups: normal group, model group, low-dose crocin group, high-dose crocin group, and positive control group. Experimental rat model of ulcerative colitis was induced by dextran sodium sulfate. Starting on the first day of modeling, rats were routinely fed in the normal group, were given sulfasalazine by gavage in the positive drug group, and were gavaged with 0.05 and 0.1 g/kg crocin in the low-and high-dose crocin groups, respectively. RESULTS AND CONCLUSION: One week after intervention, the crocin-treated rats had significantly decreased scores on colon tissue injury and Crohn’s disease activity index(P < 0.05). Compared with the model group, crocin groups had a decrease in the content of malondialdehyde and activity of myeloperoxidase(P < 0.05), and an increase in the activity of superoxide dismutase in the colon tissue(P < 0.05). Shown by immunohistochemical staining, compared with the model group, treatment with crocin significantly reduced immune responses of tumor necrosis factor α and interleukin 23 proteins after 1 week of intervention(P < 0.05). Compared with the model group, treatment with crocin downregulated the expression levels of total protein Bax, Caspase-3, Toll-like receptor 4 and MyD88(P < 0.05), and upregulated the expression of Bcl-2 in the intestinal tissue of rats(P < 0.05). These results indicate that crocin has a certain therapeutic effect in ulcerative colitis rats and its mechanism may be related to down-regulation of the Toll-like receptor 4/MyD88 signaling pathway and inhibition of oxidative stress, inflammation and apoptosis in the colon.