AIM： To investigate the roles of lymphocytes in the development of dextran sulfate sodium-induced colitis. METHODS： Using various doses of dextran sulfate sodium （DSS）, we induced colitis in wild-type B6 control a...AIM： To investigate the roles of lymphocytes in the development of dextran sulfate sodium-induced colitis. METHODS： Using various doses of dextran sulfate sodium （DSS）, we induced colitis in wild-type B6 control and Rag-1 knockout （H-2b haplotype） mice, and evaluated the colitis in terms of symptomatic and histologic parameters, such as weight loss, survival, severity of diarrhea, shortage of colon length and histological changes. Symptomatic parameters were checked daily and histological changes were scored. RESULTS： Although development of colitis in Rag-1 knockout mice treated with high dose （5%） of DSS was comparable to that in B6 control mice, colitis progression was much more tolerable in Rag-1 knockout mice compared to than in B6 mice treated with low dose （1.5%） DSS. Symptomatic parameters as well as histopathologic changes were improved in Rag-1 knockout mice. CONCLUSION： These results indicate that the presence of lymphoo/tes contributes to colitis progression at low dose of DSS stimulation. Lymphoo/tes may play roles as an aggravating factor in DSS-induced colitis.展开更多
AIM: To investigate anti-hypersensitive effects of α2δ-1 ligands in non-inflammatory and inflammationassociated colonic hypersensitivity(CHS) mouse models.METHODS: To induce an inflammation-associated CHS, 1% dextra...AIM: To investigate anti-hypersensitive effects of α2δ-1 ligands in non-inflammatory and inflammationassociated colonic hypersensitivity(CHS) mouse models.METHODS: To induce an inflammation-associated CHS, 1% dextran sulfate sodium(DSS) was administered to C57Bl/6J male mice, in drinking water, for 14 d. Regarding the non-inflammatory neonatal maternal separation(NMS)-induced CHS model, wild-type C57BI/6J pups were isolated from their mother from day 2 to day 14(P2 to P14), three hours per day(from 9:00 a.m. to 12:00 p.m.). Colorectal distension was performed by inflating distension probe from 20 μL to 100 μL by 20 μL increment step every 10 s. After a first colorectal distension(CRD), drugs were administered subcutaneously, in a cumulative manner,(Gabapentin at 30 mg/kg and 100 mg/kg; Pregabalin at 10 mg/kg and 30 mg/kg; Carbamazepine at 10 mg/kg and 30 mg/kg) and a second CRD was performed one hour after each injection.RESULTS: The visceromotor response(VMR) to CRD was increased by our NMS paradigm protocol in comparison to non-handled(NH) mice, considering the highest distension volumes(80 μL: 0.783 ± 0.056 mV /s vs 0.531 ± 0.034 m V/s, P < 0.05 and 100 μL: 1.087 ± 0.056 m V/s vs 0.634 ± 0.038 m V/s, P < 0.05 for NMS and NH mice, respectively). In the inflammationassociated CHS, DSS-treated mice showed a dramatic and significant increase in VMR at 60 and 80 μL distension volumes when compared to control mice(60 μL: 0.920 ± 0.079 m V/s vs 0.426 ± 0.100 m V/s P < 0.05 and 80 μL: 1.193 ± 0.097 mV /s vs 0.681 ± 0.094 mV /s P < 0.05 for DSS- and Water-treated mice, respectively). Carbamazepine failed to significantly reduce CHS in both models. Gabapentin significantly reduced CHS in the DSS-induced model for both subcutaneous injections at 30 or 100 mg/kg. Pregabalin s i g n i f i c a n t l y r e d u c e d V M R t o C R D i n t h e n o n-inflammatory NMS-induced CHS model for the acute subcutaneous administration of the highest cumulative dose(30 mg/kg) and significantly reduced CHS in lowdose DSS-treated mice in a dose-dependent manner. Finally, the percent decrease of AUC induced by acute GBP or Pregabalin treatment were higher in the inflammatory DSS-induced CHS model in comparison to the non-inflammatory NMS-induced CHS model.CONCLUSION: This preclinical study demonstrates α2δ-1 ligands efficacy on inflammation-associated CHS, highlighting their potential clinical interest in patients with chronic abdominal pain and moderate intestinal inflammation.展开更多
BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin co...BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin complex 1(mTORC1)activation.Previous studies have demonstrated the therapeutic effects of BT2 on arthritis,liver cancer,and kidney injury.However,the effects of BT2 on ulcerative colitis(UC)are unknown.AIM To investigate the anti-UC effects of BT2 and the underlying mechanism.METHODS Mouse UC models were created through the administration of 3.5%dextran sodium sulfate(DSS)for 7 d.The mice in the treated groups were administered salazosulfapyridine(300 mg/kg)or BT2(20 mg/kg)orally from day 1 to day 7.At the end of the study,all of the mice were sacrificed,and colon tissues were removed for hematoxylin and eosin staining,immunoblot analyses,and immunohistochemical assays.Cytokine levels were measured by flow cytometry.The contents of BCAAs including valine,leucine,and isoleucine,in mouse serum were detected by liquid chromatography-tandem mass spectrometry,and the abundance of intestinal flora was analyzed by 16S ribosomal DNA sequencing.RESULTS Our results revealed that BT2 significantly ameliorated the inflammatory symptoms and pathological damage induced by DSS in mice.BT2 also reduced the production of the proinflammatory cytokines interleukin 6(IL-6),IL-9,and IL-2 and increased the anti-inflammatory cytokine IL-10 level.In addition,BT2 notably improved BCAA catabolism and suppressed mTORC1 activation and cyclooxygenase-2 expression in the colon tissues of UC mice.Furthermore,highthroughput sequencing revealed that BT2 restored the gut microbial abundance and diversity in mice with colitis.Compared with the DSS group,BT2 treatment increased the ratio of Firmicutes to Bacteroidetes and decreased the abundance of Enterobacteriaceae and Escherichia-Shigella.CONCLUSION Our results indicated that BT2 significantly ameliorated DSS-induced UC and that the latent mechanism involved the suppression of BCAA-associated mTORC1 activation and modulation of the intestinal flora.展开更多
目的探讨预防性使用硫酸右旋糖苷(DS)和治疗性使用DS对人胃癌细胞腹腔种植的作用。方法培养胃癌BGC-823细胞,构建裸鼠动物模型:选用BALB/C裸鼠共110只,随机分为预防组(60只)和治疗组(50只)。预防组在注射肿瘤细胞的同时注射DS,治疗组注...目的探讨预防性使用硫酸右旋糖苷(DS)和治疗性使用DS对人胃癌细胞腹腔种植的作用。方法培养胃癌BGC-823细胞,构建裸鼠动物模型:选用BALB/C裸鼠共110只,随机分为预防组(60只)和治疗组(50只)。预防组在注射肿瘤细胞的同时注射DS,治疗组注射肿瘤细胞24 h后注射DS。打开裸鼠腹腔,观察胃癌细胞腹腔种植的数量,所有结节经HE染色证实。应用免疫组织化学染色(SP法)检测大网膜整合素β1蛋白的表达,应用RT-PCR方法检测大网膜整合素β1 m RNA的表达。结果 HE染色结果显示治疗组裸鼠腹腔转移的肿瘤结节数量明显多于预防组,差异有统计学意义(P<0.05),SP法结果显示治疗组整合素β1蛋白的表达量明显高于预防组,差异有统计学意义(P<0.01),RT-PCR结果显示治疗组整合素β1 m RNA的表达量明显高于预防组,差异有统计学意义(P<0.01)。结论 DS预防胃癌腹腔种植转移的效果比治疗胃癌腹腔种植转移的效果更佳。展开更多
AIM: To evaluate the efficacy of the improvedthrombospondin mimetic peptide ABT-898 in a murine model of ulcerative colitis. METHODS: The dextran sodium sulfate(DSS) was used for the induction of colitis in both TSP-1...AIM: To evaluate the efficacy of the improvedthrombospondin mimetic peptide ABT-898 in a murine model of ulcerative colitis. METHODS: The dextran sodium sulfate(DSS) was used for the induction of colitis in both TSP-1 deficient(TSP-1-/-) and wild type(WT) mice during 7 d. While mice were receiving the DSS dissolved in the drinking water, the ABT-898 peptide was dissolved in sterile 5% glucose solution and delivered using mini pumps subcutaneously implanted. Plasma samples were analyzed for interleukin(IL)-6 by ELISA assay and colonic tissues were harvested, fixed and processed for histological evaluation. Immunohistochemistry using antibodies for the detection of CD31 and MECA in endothelial cells was performed. Inflammation was graded in colonic sections and the number of microvessels in each lesion was assessed. Activation of signal transducer and activator of transcription 3(STAT3) in colonic samples was quantified by immunohistochemistry and Western blotting using antibodies against total STAT3 and phosphorylated STAT3(p STAT3)(Ser727).RESULTS: Treatment with ABT-898 considerably diminished the inflammatory response in WT and TSP-1-/- mice(P < 0.0001 in both groups vs control). Identification of blood vessels highlighted by CD31/MECA immunohistochemistry, showed significantly reduced vessel counts in colitic lesions of WT and TSP-1-/- mice treated with ABT898(TSP-1-/- controls/TSP-1-/- treated, P = 0.0002; WT controls/WT treated, P = 0.0005). Consistently, IL-6 was significantly diminished in plasma samples of TSP-1-/- and WT treated with the peptide when compared to the control mice(P = 0.0002 and P = 0.0148, respectively). p STAT3 positive cells were quantified in WT and TSP-1-/- treated with ABT-898. A significant decrease in positive cells for p STAT3 was observed in treated mice(TSP-1-/- controls/TSP-1-/- treated, P = 0.0089; WT/WT treated, P = 0.0110). These results were confirmedby Western blotting analyses showing lower levels of p STAT3 in colitic lesions from mice treated with the peptide ABT-898.CONCLUSION: These findings indicate that the new peptide ABT-898 ameliorates inflammation and angiogenesis and might be a therapeutic alternative in IBD and inflammatory diseases.展开更多
目的:观察Pim-1在炎症性肠病发病过程中的动态表达,并观察其与炎症程度的相关性。方法:BALB/c小鼠饮用5%葡聚糖硫酸钠溶液建立急性炎症性肠病模型,分别在第0、1、4、7天取结肠标本,利用real time PCR、免疫组化动态观察Pim-1表达,分析Pi...目的:观察Pim-1在炎症性肠病发病过程中的动态表达,并观察其与炎症程度的相关性。方法:BALB/c小鼠饮用5%葡聚糖硫酸钠溶液建立急性炎症性肠病模型,分别在第0、1、4、7天取结肠标本,利用real time PCR、免疫组化动态观察Pim-1表达,分析Pim-1的表达和疾病活动指数、组织学炎症评分的相关性。结果:饮用5%葡聚糖硫酸钠7天后成功建立小鼠急性炎症性肠病动物模型;Real time PCR、免疫组化结果显示在饮用5%DSS第4、7天后Pim-1表达较正常组及第1天均明显升高,P<0.05,而且Pim-1蛋白主要在淋巴细胞、中性粒细胞等炎症细胞表达;Pearson相关分析表明,结肠组织中Pim-1蛋白与疾病活动指数呈正相关(R=0.868,P<0.01),与组织病理学评分亦呈正相关(R=0.851,P<0.01)。结论:在炎症性肠病起病过程中Pim-1的表达与肠道炎症程度呈正相关,提示Pim-1信号参与肠道炎症反应。展开更多
基金Supported by Hallym University Research Fund, 2004, No.HRF-2004 -44
文摘AIM： To investigate the roles of lymphocytes in the development of dextran sulfate sodium-induced colitis. METHODS： Using various doses of dextran sulfate sodium （DSS）, we induced colitis in wild-type B6 control and Rag-1 knockout （H-2b haplotype） mice, and evaluated the colitis in terms of symptomatic and histologic parameters, such as weight loss, survival, severity of diarrhea, shortage of colon length and histological changes. Symptomatic parameters were checked daily and histological changes were scored. RESULTS： Although development of colitis in Rag-1 knockout mice treated with high dose （5%） of DSS was comparable to that in B6 control mice, colitis progression was much more tolerable in Rag-1 knockout mice compared to than in B6 mice treated with low dose （1.5%） DSS. Symptomatic parameters as well as histopathologic changes were improved in Rag-1 knockout mice. CONCLUSION： These results indicate that the presence of lymphoo/tes contributes to colitis progression at low dose of DSS stimulation. Lymphoo/tes may play roles as an aggravating factor in DSS-induced colitis.
文摘目的:探讨双歧杆菌三联活菌对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的结肠炎小鼠肠道炎症及紧密连接蛋白连接黏附分子1(junctional adhesion molecule-1,JAM-1)表达与分布的影响.方法:将SPF级♀Balb/c小鼠(8-10周龄)随机分为5组:正常对照NC组、益生菌BB组(正常小鼠予益生菌灌胃14 d)、急性DSS模型组(生理盐水灌胃14 d+第8天起4%DSS溶液自由饮用7 d)、BD组(益生菌灌胃7 d+第8天起4%D S S溶液自由饮用7 d)、B D B组(益生菌灌胃14 d+第8天起4%D S S溶液自由饮用7 d).记录小鼠每日体质量变化.第15天处死小鼠、分离结肠,观察各组肠道损伤情况,以组织病理学积分评估结肠炎的严重程度.采用Western blot法、免疫组织化学法(IHC)检测结肠组织中JAM-1的表达及分布.结果:与D S S模型组相比,益生菌干预组(BD、BDB组)小鼠减轻的体质量得到不同程度恢复(95.17%±3.34%、87.17%±1.83%vs 81.49%±2.16%,P1<0.01,P2=0.08),病变结肠缩短程度、肠黏膜水肿,上皮细胞破坏程度及肠道炎症细胞浸润减轻,且BD组小鼠的改善程度优于BDB组.Western blot结果显示给予益生菌干预,BD、B D B组肠黏膜组织JAM-1蛋白的表达水平较DSS组显著增加(0.725±0.027、0.739±0.033 vs 0.454±0.073,均P<0.05).免疫组织化学结果显示DSS模型组小鼠结肠黏膜JAM-1染色强度明显减弱,染色细胞数减少,分布散乱、中断,益生菌处理后JAM-1染色强度及阳性细胞数减少程度显著改善.仅给予益生菌灌胃的小鼠,体质量、结肠大体、镜下表现以及结肠组织中JAM-1蛋白表达水平及分布,与正常小鼠无显著差异(P>0.05).结论:双歧杆菌三联活菌可能通过影响JAM-1蛋白的表达与分布,改善肠黏膜的屏障功能,从而减轻小鼠急性期肠道炎症反应.
文摘AIM: To investigate anti-hypersensitive effects of α2δ-1 ligands in non-inflammatory and inflammationassociated colonic hypersensitivity(CHS) mouse models.METHODS: To induce an inflammation-associated CHS, 1% dextran sulfate sodium(DSS) was administered to C57Bl/6J male mice, in drinking water, for 14 d. Regarding the non-inflammatory neonatal maternal separation(NMS)-induced CHS model, wild-type C57BI/6J pups were isolated from their mother from day 2 to day 14(P2 to P14), three hours per day(from 9:00 a.m. to 12:00 p.m.). Colorectal distension was performed by inflating distension probe from 20 μL to 100 μL by 20 μL increment step every 10 s. After a first colorectal distension(CRD), drugs were administered subcutaneously, in a cumulative manner,(Gabapentin at 30 mg/kg and 100 mg/kg; Pregabalin at 10 mg/kg and 30 mg/kg; Carbamazepine at 10 mg/kg and 30 mg/kg) and a second CRD was performed one hour after each injection.RESULTS: The visceromotor response(VMR) to CRD was increased by our NMS paradigm protocol in comparison to non-handled(NH) mice, considering the highest distension volumes(80 μL: 0.783 ± 0.056 mV /s vs 0.531 ± 0.034 m V/s, P < 0.05 and 100 μL: 1.087 ± 0.056 m V/s vs 0.634 ± 0.038 m V/s, P < 0.05 for NMS and NH mice, respectively). In the inflammationassociated CHS, DSS-treated mice showed a dramatic and significant increase in VMR at 60 and 80 μL distension volumes when compared to control mice(60 μL: 0.920 ± 0.079 m V/s vs 0.426 ± 0.100 m V/s P < 0.05 and 80 μL: 1.193 ± 0.097 mV /s vs 0.681 ± 0.094 mV /s P < 0.05 for DSS- and Water-treated mice, respectively). Carbamazepine failed to significantly reduce CHS in both models. Gabapentin significantly reduced CHS in the DSS-induced model for both subcutaneous injections at 30 or 100 mg/kg. Pregabalin s i g n i f i c a n t l y r e d u c e d V M R t o C R D i n t h e n o n-inflammatory NMS-induced CHS model for the acute subcutaneous administration of the highest cumulative dose(30 mg/kg) and significantly reduced CHS in lowdose DSS-treated mice in a dose-dependent manner. Finally, the percent decrease of AUC induced by acute GBP or Pregabalin treatment were higher in the inflammatory DSS-induced CHS model in comparison to the non-inflammatory NMS-induced CHS model.CONCLUSION: This preclinical study demonstrates α2δ-1 ligands efficacy on inflammation-associated CHS, highlighting their potential clinical interest in patients with chronic abdominal pain and moderate intestinal inflammation.
基金Supported by National Natural Science Foundation of ChinaNo. 82074241+1 种基金Project of Jiangsu Province Hospital of Traditional Chinese Medicine Peak TalentNo. y2021rc36
文摘BACKGROUND 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid(BT2)is a benzothiophene carboxylate derivative that can suppress the catabolism of branched-chain amino acid(BCAA)-associated mammalian target of rapamycin complex 1(mTORC1)activation.Previous studies have demonstrated the therapeutic effects of BT2 on arthritis,liver cancer,and kidney injury.However,the effects of BT2 on ulcerative colitis(UC)are unknown.AIM To investigate the anti-UC effects of BT2 and the underlying mechanism.METHODS Mouse UC models were created through the administration of 3.5%dextran sodium sulfate(DSS)for 7 d.The mice in the treated groups were administered salazosulfapyridine(300 mg/kg)or BT2(20 mg/kg)orally from day 1 to day 7.At the end of the study,all of the mice were sacrificed,and colon tissues were removed for hematoxylin and eosin staining,immunoblot analyses,and immunohistochemical assays.Cytokine levels were measured by flow cytometry.The contents of BCAAs including valine,leucine,and isoleucine,in mouse serum were detected by liquid chromatography-tandem mass spectrometry,and the abundance of intestinal flora was analyzed by 16S ribosomal DNA sequencing.RESULTS Our results revealed that BT2 significantly ameliorated the inflammatory symptoms and pathological damage induced by DSS in mice.BT2 also reduced the production of the proinflammatory cytokines interleukin 6(IL-6),IL-9,and IL-2 and increased the anti-inflammatory cytokine IL-10 level.In addition,BT2 notably improved BCAA catabolism and suppressed mTORC1 activation and cyclooxygenase-2 expression in the colon tissues of UC mice.Furthermore,highthroughput sequencing revealed that BT2 restored the gut microbial abundance and diversity in mice with colitis.Compared with the DSS group,BT2 treatment increased the ratio of Firmicutes to Bacteroidetes and decreased the abundance of Enterobacteriaceae and Escherichia-Shigella.CONCLUSION Our results indicated that BT2 significantly ameliorated DSS-induced UC and that the latent mechanism involved the suppression of BCAA-associated mTORC1 activation and modulation of the intestinal flora.
文摘目的探讨预防性使用硫酸右旋糖苷(DS)和治疗性使用DS对人胃癌细胞腹腔种植的作用。方法培养胃癌BGC-823细胞,构建裸鼠动物模型:选用BALB/C裸鼠共110只,随机分为预防组(60只)和治疗组(50只)。预防组在注射肿瘤细胞的同时注射DS,治疗组注射肿瘤细胞24 h后注射DS。打开裸鼠腹腔,观察胃癌细胞腹腔种植的数量,所有结节经HE染色证实。应用免疫组织化学染色(SP法)检测大网膜整合素β1蛋白的表达,应用RT-PCR方法检测大网膜整合素β1 m RNA的表达。结果 HE染色结果显示治疗组裸鼠腹腔转移的肿瘤结节数量明显多于预防组,差异有统计学意义(P<0.05),SP法结果显示治疗组整合素β1蛋白的表达量明显高于预防组,差异有统计学意义(P<0.01),RT-PCR结果显示治疗组整合素β1 m RNA的表达量明显高于预防组,差异有统计学意义(P<0.01)。结论 DS预防胃癌腹腔种植转移的效果比治疗胃癌腹腔种植转移的效果更佳。
基金Supported by National Institutes of Health(AREA),No.R15 DK067901-02the Howard Hughes Medical Institute,No.52006328Wilkes Mentoring funds and institutional funds from Wilkes University and TCMC
文摘AIM: To evaluate the efficacy of the improvedthrombospondin mimetic peptide ABT-898 in a murine model of ulcerative colitis. METHODS: The dextran sodium sulfate(DSS) was used for the induction of colitis in both TSP-1 deficient(TSP-1-/-) and wild type(WT) mice during 7 d. While mice were receiving the DSS dissolved in the drinking water, the ABT-898 peptide was dissolved in sterile 5% glucose solution and delivered using mini pumps subcutaneously implanted. Plasma samples were analyzed for interleukin(IL)-6 by ELISA assay and colonic tissues were harvested, fixed and processed for histological evaluation. Immunohistochemistry using antibodies for the detection of CD31 and MECA in endothelial cells was performed. Inflammation was graded in colonic sections and the number of microvessels in each lesion was assessed. Activation of signal transducer and activator of transcription 3(STAT3) in colonic samples was quantified by immunohistochemistry and Western blotting using antibodies against total STAT3 and phosphorylated STAT3(p STAT3)(Ser727).RESULTS: Treatment with ABT-898 considerably diminished the inflammatory response in WT and TSP-1-/- mice(P < 0.0001 in both groups vs control). Identification of blood vessels highlighted by CD31/MECA immunohistochemistry, showed significantly reduced vessel counts in colitic lesions of WT and TSP-1-/- mice treated with ABT898(TSP-1-/- controls/TSP-1-/- treated, P = 0.0002; WT controls/WT treated, P = 0.0005). Consistently, IL-6 was significantly diminished in plasma samples of TSP-1-/- and WT treated with the peptide when compared to the control mice(P = 0.0002 and P = 0.0148, respectively). p STAT3 positive cells were quantified in WT and TSP-1-/- treated with ABT-898. A significant decrease in positive cells for p STAT3 was observed in treated mice(TSP-1-/- controls/TSP-1-/- treated, P = 0.0089; WT/WT treated, P = 0.0110). These results were confirmedby Western blotting analyses showing lower levels of p STAT3 in colitic lesions from mice treated with the peptide ABT-898.CONCLUSION: These findings indicate that the new peptide ABT-898 ameliorates inflammation and angiogenesis and might be a therapeutic alternative in IBD and inflammatory diseases.
文摘目的:观察Pim-1在炎症性肠病发病过程中的动态表达,并观察其与炎症程度的相关性。方法:BALB/c小鼠饮用5%葡聚糖硫酸钠溶液建立急性炎症性肠病模型,分别在第0、1、4、7天取结肠标本,利用real time PCR、免疫组化动态观察Pim-1表达,分析Pim-1的表达和疾病活动指数、组织学炎症评分的相关性。结果:饮用5%葡聚糖硫酸钠7天后成功建立小鼠急性炎症性肠病动物模型;Real time PCR、免疫组化结果显示在饮用5%DSS第4、7天后Pim-1表达较正常组及第1天均明显升高,P<0.05,而且Pim-1蛋白主要在淋巴细胞、中性粒细胞等炎症细胞表达;Pearson相关分析表明,结肠组织中Pim-1蛋白与疾病活动指数呈正相关(R=0.868,P<0.01),与组织病理学评分亦呈正相关(R=0.851,P<0.01)。结论:在炎症性肠病起病过程中Pim-1的表达与肠道炎症程度呈正相关,提示Pim-1信号参与肠道炎症反应。
