1.Determination of DNA-DNA Hybridization Condition for Rapid Identification of Mycobacterium Species.
Yun Sop CHONG ; Sang Nae CHO ; Kyung Won LEE ; Hong Seok PARK
Journal of the Korean Society for Microbiology 1999;34(2):137-145
Rapid identification of Mycobacterium spp. isolated from patients is important with increased isolation of mycobacteria other than tubercle bacilli (MOTT). DNA-DNA hybridization with streptavidin-peroxidase and tetramethylbenzidine (TMB) color reaction method was recognized as a useful tool for identification of various species of mycobacteria. In this study, optimum condition of the test was determined. The optimal concentrations of tetramethylbenzidine dihydrochloride and hydrogen peroxide for streptavidin-horseradish peroxidase were 0.3-0.6 ug/ ml and 0.16 mM, respectively. The TMB stock solution was stable when prepared in methanol and the dilution of TBM stock solution in 10 mM sodium citrate-10 mM EDTA solution (pH 5.0) gave highest peroxidase-TMB activity. The suitable composition of hybridization solution consisted of 2 x SSC, 10% dextran sulfate, 50 ug/ml salmon DNA, 5 x Denhardt's solution, and 50% formamide. The 5-minute heating at 100C of test DNA prior to photobiotin labeling significantly increased the reaction. In conclusion, DNA-DNA hybridization method with streptavidin-peroxidase and TMB color reaction method may be useful for rapid identification of Mycobacterium spp. isolated from patients.
Dextran Sulfate
;
DNA
;
Edetic Acid
;
Heating
;
Hot Temperature
;
Humans
;
Hydrogen Peroxide
;
Methanol
;
Mycobacterium*
;
Peroxidase
;
Salmon
;
Sodium
2.Kirenol relieves dextran sulfate sodium-induced ulcerative colitis in mice by inhibiting inflammatory cytokines and inducing CD4 T lymphocyte apoptosis.
Xiuhong LIU ; D U YAJUN ; Guoxing LIU ; Guomei DAN ; Xin TONG ; Juan XIAO
Journal of Southern Medical University 2019;39(12):1387-1392
OBJECTIVE:
To investigate whether kirenol, the major pharmacologically active compound of the Chinese medicinal herb , can protect mice from dextran sulfate sodium (DSS)-induced ulcerative colitis (UC).
METHODS:
C57BL/6 mice with or without kirenol pretreatment were treated with DSS in drinking water for 7 days to induce UC. The symptoms of UC including weight loss, diarrhea and bloody stool were observed daily and graded using the disease activity index (DAI). Colon injury of the mice was assessed by measuring the length of the colon and HE staining of the colon tissue. The levels of inflammatory cytokines produced by the mesenteric lymph nodes (MLNs) lymphocytes were measured using enzyme-linked immunosorbent assay; the apoptosis of the lymphocytes and CD4 T cells was analyzed using flow cytometry.
RESULTS:
The mice receiving pretreatment with kirenol showed obviously ameliorated symptoms of UC and milder pathological changes in the colon as compared with the control mice. Kirenol treatment significantly down-regulated the secretion of IFN-γ, IL-17A, IL-6 and TNF-α by the MLNs lymphocytes and increased the apoptosis of lymphocytes, especially CD4 T cells in the DSS-treated mice.
CONCLUSIONS
Kirenol can protect against T cell-mediated colon injury in DSS-treated mice possibly by suppressing the secretion of inflammatory mediators and inducing apoptosis of the inflammatory lymphocytes.
Animals
;
Apoptosis
;
Colitis, Ulcerative
;
Cytokines
;
Dextran Sulfate
;
Diterpenes
;
Mice
;
Mice, Inbred C57BL
;
T-Lymphocytes
3.Effect of Aging on Experimental Colitis in Mice and Its Associated Mechanism.
Ai Ling LIU ; Hong Ying WANG ; Hong LÜ ; Jia Ming QIAN
Acta Academiae Medicinae Sinicae 2019;41(1):28-36
Objective To explore whether aging increases severity of colitis in mice and its mechanism.Methods Young (6-8 weeks)and aged (56 weeks) C57Bl/6 mice were divided into the control and experimental group (n=5,each). Dextran sodium sulfate(DSS) was used to induce acute colitis mouse model in the experimental group.The mRNA expressions of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in colon were measured by RT-PCR. Tight junctions (TJs) of intestinal epithelial cells was examined by transmission electron microscopy (TEM). Protein expressions of E-cadherin and occludin were detected by Western blotting and immunohistochemistry in colon.Results Compared with the young DSS-induced mice,the aged DSS-induced mice had more weight loss(t=3.679,P=0.006),higher disease indexes (t=2.496,P=0.037),higher histologic scores(U=0.000,P=0.008) and higher colonic IL-6 level (U=4.000,P=0.191). The TJs of intestinal epithelial cells were discontinuous in old healthy rats,and the TJs were destroyed significantly in both young and aged DSS-induced mice. Compared with the young DSS-induced mice,the aged DSS-induced mice had decreased protein expressions of E-cadherin (t=0.184,P=0.863)and occludin (t=0.399,P=0.710).Conclusions Aging leads to more severe disease following DSS challenge. Age-related deterioration in the functions of the gastrointestinal barrier and integrity may be one of the possible mechanisms.
Animals
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Colitis
;
Colon
;
Dextran Sulfate
;
Disease Models, Animal
;
Intestinal Mucosa
;
Mice
;
Mice, Inbred C57BL
;
Rats
4.Therapeutic effect of the immunomodulatory drug lenalidomide, but not pomalidomide, in experimental models of rheumatoid arthritis and inflammatory bowel disease.
Belen LOPEZ-MILLAN ; Rafael DIAZ DE LA GUARDIA ; Heleia ROCA-HO ; Carmen M GARCÍA-HERRERO ; Jessie R LAVOIE ; Michael ROSU-MYLES ; Elena GONZALEZ-REY ; Francisco O'VALLE ; Gabriel CRIADO ; Mario DELGADO ; Pablo MENENDEZ
Experimental & Molecular Medicine 2017;49(2):e290-
Thalidomide is an immunomodulatory drug (IMiD) with proven therapeutic action in several autoimmune/inflammatory diseases; however, its inherent high toxicity has led to the development of more powerful and safer thalidomide analogs, including lenalidomide and pomalidomide. These are new generation IMiDs that exhibit direct antitumor activity as well as anti-inflammatory/immunomodulatory properties, and are FDA-approved for the treatment of several hematological malignances. Here we investigated the potential therapeutic effects of lenalidomide and pomalidomide in several experimental murine models of autoimmune/inflammatory diseases: 2,4,6-trinitrobenzene sulfonic acid- and dextran sulfate sodium-induced inflammatory bowel disease and type II collagen-induced arthritis. Lenalidomide displayed a strong therapeutic effect in all these models of autoimmune/inflammatory diseases, while the effect of pomalidomide was less pronounced. In vitro experiments confirmed the immunosuppressive effect of both IMiDs on the proliferative response of stimulated human lymphocytes and on the balance of secreted cytokines toward an anti-inflammatory profile. We conclude that lenalidomide may offer a therapeutic opportunity against autoimmune/inflammatory diseases.
Arthritis, Experimental
;
Arthritis, Rheumatoid*
;
Cytokines
;
Dextran Sulfate
;
Humans
;
In Vitro Techniques
;
Inflammatory Bowel Diseases*
;
Lymphocytes
;
Models, Theoretical*
;
Thalidomide
;
Therapeutic Uses
5.Histological Study of Experimental Colitis Induced by Dextran Sulfate Sodium.
Korean Journal of Anatomy 2006;39(1):27-33
Inflamatory bowel disease is the general medical terminology on chronic inflammatory illness of unknown origin, but it is considered that environmental, genetical, immunological factors may develop this chronic disease. I examined the histological changes on the experimental ulcerative colitis induced by dextran sulfate sodium(DSS) in rats. Experimental colitis was induced with 5% DSS in the drinking water for 5days in rats (experimental group). And the repair group were treated with 5% DSS for 5days and with pure water after 7days in rats. In experimental group, there are many inflammatory finding in colon of rat which contained loss of body weight, crypt erosion, recruitment of inflammatory cells, submucosal edema. In repair group, the inflammation was recovered that the body weight was incerased, the crypt was recovered. And Ki 67 immunoreaction were restricted lower 1/3 crypt in normal group but positive Ki 67 reaction appeared in the repaired all region. In this study, DSS was induced experimental colitis and the colitis was repaired when we stoped DSS supply. And the Ki 67 during repaired period were overproduction.
Animals
;
Body Weight
;
Chronic Disease
;
Colitis*
;
Colitis, Ulcerative
;
Colon
;
Dextran Sulfate*
;
Dextrans*
;
Drinking Water
;
Edema
;
Immunologic Factors
;
Inflammation
;
Rats
;
Water
6.Effect of Nrf2/HO-1 signaling pathway in intestinal protection by Sishen Pills against ulcerative colitis in mice.
Xue-Xia ZHANG ; Jian-Wen JIN ; Chang-He LIU ; Min ZHOU ; Ying-Xin HE ; Fei WANG ; Fang-Zhou LIU
China Journal of Chinese Materia Medica 2021;46(16):4187-4192
The present study aimed to explore the effect of nuclear factor erythroid 2 related factor 2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway in intestinal protection by Sishen Pills against ulcerative colitis(UC). After the UC model was induced by 3% dextran sodium sulfate(DSS), experimental animals were randomly divided into control group, model group, salazosulfapyridine(SASP) group, and low-and high-dose Sishen Pills groups. Drug intervention(ig) was performed for seven consecutive days during modeling. On the 7 th day, the mice were euthanized. The body weight and colon length were recorded, and the histopathological changes of the colon were observed by HE staining. Serum interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), total antioxidant capacity(T-AOC), malondialdehyde(MDA), and reactive oxygen species(ROS) were detected by ELISA. The protein and mRNA expression of Nrf2, HO-1, and NADPH quinine oxidoreductase-1(NQO-1) was determined by Western blot and reverse transcription-polymerase chain reaction(RT-PCR). Compared with the normal group, the model group exhibited reduced body weight, colon length, and T-AOC, increased IL-6, TNF-α, MDA, and ROS, and diminished protein and mRNA expression of Nrf2, HO-1, and NQO-1 in the colon tissues. Compared with the model group, the SASP group and high-dose Sishen Pills group showed elevated body weight, colon length, and T-AOC, lowered IL-6, TNF-α, MDA, and ROS levels, and increased protein and mRNA expression of Nrf2, HO-1, and NQO-1 in the colon tissues. As assessed by HE staining, Sishen Pills could improve the pathological changes of the colon. The findings suggested that Sishen Pills could protect the colon against UC induced by 3% DSS. The specific mechanism of action may be related to the anti-inflammatory and anti-oxidative stress effects by the activation of the Nrf2/HO-1 signaling pathway.
Animals
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Colitis, Ulcerative/genetics*
;
Dextran Sulfate
;
Heme Oxygenase-1/metabolism*
;
Mice
;
NF-E2-Related Factor 2/metabolism*
;
Signal Transduction
7.Fermented Pueraria Lobata extract ameliorates dextran sulfate sodium-induced colitis by reducing pro-inflammatory cytokines and recovering intestinal barrier function.
Seungho CHOI ; Jong Kyu WOO ; Yeong Su JANG ; Ju Hee KANG ; Jung Eun JANG ; Tae Hoo YI ; Sang Yong PARK ; Sun Yeou KIM ; Yeo Sung YOON ; Seung Hyun OH
Laboratory Animal Research 2016;32(3):151-159
Inflammatory bowel disease is a chronic inflammatory disorder occurring in the gastrointestinal track. However, the efficacy of current therapeutic strategies has been limited and accompanied by side effects. In order to eliminate the limitations, herbal medicines have recently been developed for treatment of IBD. Peuraria Lobata (Peuraria L.) is one of the traditional herbal medicines that have anti-inflammatory effects. Bioavailability of Peuraria L., which is rich in isoflavones, is lower than that of their fermented forms. In this study, we generated fermented Peuraria L. extracts (fPue) and investigated the role of fPue in inflammation and intestinal barrier function in vitro and in vivo. As the mice or intestinal epithelial cells were treated with DSS/fPue, mRNA expression of pro-inflammatory cytokines was reduced and the architecture and expression of tight junction proteins were recovered, compared to the DSS-treated group. In summary, fPue treatment resulted in amelioration of DSS-induced inflammation in the colon, and the disrupted intestinal barrier was recovered as the expression and architecture of tight junction proteins were retrieved. These results suggest that use of fPue could be a new therapeutic strategy for treatment of IBD.
Animals
;
Biological Availability
;
Colitis*
;
Colon
;
Cytokines*
;
Dextran Sulfate*
;
Dextrans*
;
Epithelial Cells
;
In Vitro Techniques
;
Inflammation
;
Inflammatory Bowel Diseases
;
Isoflavones
;
Mice
;
Pueraria*
;
RNA, Messenger
;
Tight Junction Proteins
8.Animal Models of Inflammatory Bowel Disease.
Intestinal Research 2008;6(1):8-18
Inflammatory bowel disease (IBD) is a chronic inflammatory disease of unknown etiology that includes two main disease entities-ulcerative colitis and Crohn's disease. Although the pathogenesis of IBD remains unclear, it is widely accepted that genetic, environmental and immunological factors are involved. Animal models of IBD are indispensable for the understanding of the pathogenesis and novel therapeutic applications for IBD. IBD animal models can be divided into several different categories, including models of spontaneous colitis (cotton-top tamarin colitis); inducible forms of colitis (using acetic acid, dextran sulfate sodium and indomethacin); an adoptive transfer model (CD45RB(high) transfer model); genetically engineered models (with IL-10 knockout or TCR-alpha chain knockout mice). However, there is no 'perfect' model for human disease. Investigators must make judicious choices when selecting a model for a particular study. In this review, an overview of the different IBD animal models is provided and the contribution of the models to the current understanding of disease mechanisms is discussed, with the ultimate goal to develop future therapeutic trials.
Acetic Acid
;
Adoptive Transfer
;
Animals
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Colitis
;
Crohn Disease
;
Dextran Sulfate
;
Disease Models, Animal
;
Humans
;
Immunologic Factors
;
Inflammatory Bowel Diseases
;
Interleukin-10
;
Models, Animal
;
Research Personnel
9.Alloferon Alleviates Dextran Sulfate Sodium-induced Colitis.
Hyemin KIM ; Jong Pil IM ; Joo Sung KIM ; Jae Seung KANG ; Wang Jae LEE
Immune Network 2015;15(3):135-141
Dysfunction of gut immune regulation is involved in mucosal damage in inflammatory bowel disease (IBD). However, there is still no efficacious immune-regulator for the treatment of IBD. Alloferon is a novel immune-modulatory peptide that was originally isolated from infected insects. It shows anti-inflammatory effects by the regulation of cytokine production by immune cells and their activities. Therefore, we investigated the effect of alloferon in a mouse model of colitis using dextran sulfate sodium (DSS). Colitis was induced by administration of DSS in drinking water for 7 consecutive days. It was confirmed by the presence of weight loss, diarrhea, hematochezia, and colon contraction. Alloferon was injected 4 days after DSS administration. We found that alloferon improved the pathogenesis of IBD based on the reduced disease activity index (DAI) and colon contraction. Edema, epithelial erosion, and immune cell infiltration were found in mice administered DSS, but the phenomena were reduced following alloferon treatment. The plasma level of IL-6, a classical pro-inflammatory cytokine in colitis, was also decreased by alloferon. Moreover, alloferon inhibited the TNF-alpha-induced degradation and phosphorylation of IkappaB in Colo205 colon cancer cells. Taken together, these results show that alloferon has anti-inflammatory effects and attenuates DSS-induced colitis.
Animals
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Colitis*
;
Colon
;
Colonic Neoplasms
;
Dextran Sulfate*
;
Diarrhea
;
Drinking Water
;
Edema
;
Gastrointestinal Hemorrhage
;
Inflammatory Bowel Diseases
;
Insects
;
Interleukin-6
;
Mice
;
Phosphorylation
;
Plasma
;
Weight Loss
10.Long-Term Effects of Bone Marrow-Derived Mesenchymal Stem Cells in Dextran Sulfate Sodium-Induced Murine Chronic Colitis.
Hyun Jung LEE ; Sun Hee OH ; Hui Won JANG ; Ji Hee KWON ; Kyoung Jin LEE ; Chung Hee KIM ; Soo Jung PARK ; Sung Pil HONG ; Jae Hee CHEON ; Tae Il KIM ; Won Ho KIM
Gut and Liver 2016;10(3):412-419
BACKGROUND/AIMS: Bone marrow-derived mesenchymal stem cells (BM-MSCs) have shown beneficial effects in experimental colitis models, but the underlying mechanisms are not fully understood. We investigated the long-term effects of BM-MSCs, particularly in mice with chronic colitis. METHODS: Chronic colitis was induced by administering 3% dextran sulfate sodium (DSS) in a series of three cycles. BM-MSCs were injected intravenously into DSS-treated mice three times during the first cycle. On day 33, the therapeutic effects were evaluated with clinicopathologic profiles and histological scoring. Inflammatory mediators were measured with real-time polymerase chain reaction. RESULTS: Systemic infusion of BM-MSCs ameliorated the severity of colitis, and body weight restoration was significantly promoted in the BM-MSC-treated mice. In addition, BM-MSC treatment showed a sustained beneficial effect throughout the three cycles. Microscopic examination revealed that the mice treated with BM-MSCs had fewer inflammatory infiltrates, a lesser extent of inflammation, and less crypt structure damage compared with mice with DSS-induced colitis. Anti-inflammatory cytokine levels of interleukin-10 were significantly increased in the inflamed colons of BM-MSC-treated mice compared with DSS-induced colitis mice. CONCLUSIONS: Systemic infusion of BM-MSCs at the onset of disease exerted preventive and rapid recovery effects, with long-term immunosuppressive action in mice with repeated DSS-induced chronic colitis.
Animals
;
Body Weight
;
Bone Marrow
;
Colitis*
;
Colon
;
Dextran Sulfate*
;
Dextrans*
;
Inflammation
;
Inflammatory Bowel Diseases
;
Interleukin-10
;
Mesenchymal Stromal Cells*
;
Mice
;
Real-Time Polymerase Chain Reaction