A growing body of evidence suggests that tight junction(TJ)proteins play a crucial role in the pathogenesis of various diseases,including gastrointestinal(GI)cancer and hepatocellular carcinoma(HCC).TJ proteins primar...A growing body of evidence suggests that tight junction(TJ)proteins play a crucial role in the pathogenesis of various diseases,including gastrointestinal(GI)cancer and hepatocellular carcinoma(HCC).TJ proteins primarily maintain the epithelial and endothelial cells intact together through integral proteins however,recent reports suggest that they also regulate gene expression necessary for cell proliferation,angiogenesis,and metastasis through adapter proteins such as zonula occludens(ZO).ZO proteins are membrane-associated cytosolic scaffolding proteins that modulate cell proliferation by interacting with several transcription factors.Reduced ZO proteins in GI cancer and HCC are correlated with tumor development and poor prognosis.Pubmed has searched for using the keyword ZO and gastric cancer,ZO and cancer,and ZO and HCC for the last ten years to date.This review summarized the role of ZO proteins in cell proliferation and their expression in GI cancer and HCC.Furthermore,therapeutic interventions targeting ZO in GI and liver cancers are reviewed.展开更多
AIM:To study the effect of salvianolate on tight junctions(TJs) and zonula occludens protein 1(ZO-1) in small intestinal mucosa of cirrhotic rats.METHODS:Cirrhosis was induced using carbon tetrachloride.Rats were rand...AIM:To study the effect of salvianolate on tight junctions(TJs) and zonula occludens protein 1(ZO-1) in small intestinal mucosa of cirrhotic rats.METHODS:Cirrhosis was induced using carbon tetrachloride.Rats were randomly divided into the untreated group,low-dose salvianolate(12 mg/kg) treatment group,medium-dose salvianolate(24 mg/kg) treatment group,and high-dose salvianolate(48 mg/kg) treatment group,and were treated for 2 wk.Another 10 healthy rats served as the normal control group.Histological changes in liver tissue samples were observed under a light microscope.We evaluated morphologic indices of ileal mucosa including intestinal villi width and thickness of mucosa and intestinal wall using a pathological image analysis system.Ultrastructural changes in small intestinal mucosa were investigated in the five groups using transmission electron microscopy.The changes in ZO-1 expression,a tight junction protein,were analyzed by immunocytochemistry.The staining index was calculated as the product of the staining intensity score and the proportion of positive cells.RESULTS:In the untreated group,hepatocytes showed a disordered arrangement,fatty degeneration was extensive,swelling was obvious,and disorganized lobules were divided by collagen fibers in hepatic tissue,which were partly improved in the salvianolate treated groups.In the untreated group,abundant lymphocytes infiltrated the fibrous tissue with proliferation of bile ducts,and collagen fibers gradually decreased and damaged hepatic lobules were partly repaired following salvianolate treatment.Compared with the untreated group,no differences in intestinal villi width between the five groups were observed.The villi height as well as mucosa and intestinal wall thickness gradually thickened with salvianolate treatment and were significantly shorter in the untreated group compared with those in the salvianolate treatment groups and normal group(P < 0.01).The number of microvilli decreased and showed irregular lengths and arrangements in the untreated group.The intercellular space between epithelial cells was wider.The TJs were discontinuous,which indicated disruption in TJ morphology in the untreated group.In the treated groups,the microvilli in the intestinal epithelium were regular and the TJs were gradually integrated and distinct.The expression of ZO-1 decreased in the small intestine of the untreated cirrhotic rats.The high expression rate of ZO-1 in ileal mucosa in the untreated group was significantly lower than that in the medium-dose salvianolate group(21.43% vs 64.29%,χ 2 = 5.25,P < 0.05),high-dose salvianolate group(21.43% vs 76.92%,χ 2 = 8.315,P < 0.01) and normal group(21.43% vs 90%,χ 2 = 10.98,P < 0.01).CONCLUSION:Salvianolate improves liver histopathological changes,repairs intestinal mucosa and TJ structure,and enhances ZO-1 expression in the small intestinal mucosa in cirrhotic rats.展开更多
AIM To investigate the interaction of Zot withmicrotubule.METHODS Zot affinity column was applied topurify Zot-binding protein（s）from crudeintestinal cell lysates.After incubation at roomtemperature,the column was w...AIM To investigate the interaction of Zot withmicrotubule.METHODS Zot affinity column was applied topurify Zot-binding protein（s）from crudeintestinal cell lysates.After incubation at roomtemperature,the column was washed and theproteins bound to the Zot affinity column wereeluted by step gradient with NaCl(0.3 mol·L<sup>-1</sup>-0.5mol·L<sup>-1</sup>).The fractions were subjected to6.0%-15.0%（w/v）gradient SDS-PAGE andthen transferred to PVDF membrane for N-terminal sequencing.Purified Zot and tauprotein were blotted by using anti-Zot or anti-tauantibodies.Finally,purified Zot was tested in anin vitro tubulin binding assay.RESULTS Fractions from Zot affinity columnyielded two protein bands with a Mr of 60 kU and45kU respectively.The N-terminal sequence ofthe 60 kU band resulted identical to β-tubulin.Zot also cross-reacts with anti-tau antibodies.Inthe in vitro tubulin binding assay,Zot co-precipitate with Mt,further suggesting that Zotpossesses tubulin-binding properties.CONCLUSION Taken together,these resultssuggest that Zot regulates the permeability ofintestinal tight junctions by binding tointracellular Mt,with the subsequent activationof the intracellular signaling leading to thepermeabilization of intercellular tight junctions.展开更多
Objective：To determine the relationship of thick greasy tongue fur formation and permeability of vascular endothelial cells（ECs）with the protein expression of zonula occludens-1（ZO-1）.Methods：Sprague Dawley rats...Objective：To determine the relationship of thick greasy tongue fur formation and permeability of vascular endothelial cells（ECs）with the protein expression of zonula occludens-1（ZO-1）.Methods：Sprague Dawley rats were randomly divided into a model group of severe acute pancreatitis（SAP）and a sham-operated （SO）group.The SAP rats were further divided into two subgroups on the basis of tongue-coating status：a thick greasy tongue fur group（SAP-TGF）and a normal tongue fur group（SAP-NF）.Six rats were chosen randomly from every group mentioned above for an Evans blue assay 5 days after model establishment.For the histomorphology analysis,the expressions of ZO-1 protein and mRNA were studied by hematoxylin-eosin （HE）staining,transmission electron microscope,Western blot,and Q-PCR using blood and tongue tissues, which were collected from 8 rats randomly chosen from each group.Results：The papillae density of the rat tongue surface and the caryocinesis frequency of the basal layer were significantly increased in the SAP-TGF group compared with the SO group（P0.05）.Evans blue levels in the tongue tissue of the SAP-TGF group were significantly higher than that of the SO and SAP-NF groups（P0.05）.Vascular ECs were wider and obviously fissured in the SAP-TGF group under transmission electron microscope observation.The protein and mRNA expression of ZO-1 in the SAP-TGF group were lower than those in the SAP-NF（P0.05）.Conclusions： Reproductive activity enhancement of glossal epithelial cells was one of the main characteristics of thick greasy tongue fur formation.An increase in vasopermeability was closely associated with thick greasy tongue fur formation.Tight junction structural variation of vascular ECs might play an important role in the pathological and physiological process of thick greasy tongue fur formation.展开更多
目的研究血清紧密连接蛋白闭锁小带蛋白1(zonula occludens 1,ZO-1)、咬合蛋白(occludin)、闭合蛋白-1(claudin-1)联合急性生理学与慢性健康状况Ⅱ(acute physiology and chronic health evaluationⅡ,APACHEⅡ)评分对高甘油三酯血症型...目的研究血清紧密连接蛋白闭锁小带蛋白1(zonula occludens 1,ZO-1)、咬合蛋白(occludin)、闭合蛋白-1(claudin-1)联合急性生理学与慢性健康状况Ⅱ(acute physiology and chronic health evaluationⅡ,APACHEⅡ)评分对高甘油三酯血症型重症急性胰腺炎(hypertriglyceridemia severe acute pancreatitis,HTG-SAP)并发急性肾损伤(acute kidney injury,AKI)的预测价值。方法选择2021年1月~2024年3月期间收治的96例HTG-SAP患者进行回顾性研究,根据是否并发AKI分为AKI组(n=38)和非AKI组(n=58)。比较两组临床资料及血清ZO-1、occludin、claudin-1水平的差异,筛选差异有统计学意义的因素作为自变量进行logistic回归分析并构建预测模型,通过受试者工作特征曲线分析各个指标对HTG-SAP并发AKI的预测价值。结果AKI组的APACHEⅡ评分、全身炎症反应综合征评分、甘油三酯、血肌酐、血尿素氮水平高于非AKI组,血清ZO-1、occludin、claudin-1水平低于非AKI组,差异有统计学意义(P<0.05);logistic回归分析显示,APACHEⅡ评分(OR=1.781,95%CI:1.065~2.981)升高以及血清ZO-1(0.956,95%CI:0.918~0.995)、occludin(0.967,95%CI:0.941~0.995)、claudin-1(0.193,95%CI:0.049~0.752)水平降低是HTG-SAP患者并发AKI的危险因素(P<0.05)。APACHEⅡ评分及血清ZO-1、occludin、claudin-1单独及联合模型预测HTG-SAP患者并发AKI的曲线下面积分别为0.821(95%CI:0.740~0.902)、0.852(95%CI:0.777~0.927)、0.877(95%CI:0.807~0.947)、0.936(95%CI:0.891~0.981)、0.998(95%CI:0.993~1.000)。结论血清紧密连接蛋白ZO-1、occludin、claudin-1水平降低及APACHEⅡ评分增加是HTG-SAP患者并发AKI的危险因素,根据四项指标建立预测模型对HTG-SAP患者并发AKI具有较好的预测价值。展开更多
●AIM:To evaluate the role of semaphorin 7A(Sema7A)and its associated regulatory mechanisms in modulating the barrier function of cultured human corneal epithelial cells(HCEs).●METHODS:Barrier models of HCEs were tre...●AIM:To evaluate the role of semaphorin 7A(Sema7A)and its associated regulatory mechanisms in modulating the barrier function of cultured human corneal epithelial cells(HCEs).●METHODS:Barrier models of HCEs were treated with recombinant human Sema7A at concentrations of 0,125,250,or 500 ng/mL for 24,48,or 72h in vitro.Transepithelial electrical resistance(TEER)as well as Dextran-fluorescein isothiocyanate(FITC)permeability assays were conducted to assess barrier function.To quantify tight junctions(TJs)such as occludin and zonula occludens-1(ZO-1)at the mRNA level,reverse transcriptionpolymerase chain reaction(RT-PCR)analysis was performed.Immunoblotting was used to examine the activity of the nuclear factor-kappa B(NF-κB)signaling pathway and the production of TJs proteins.Immunofluorescence analyses were employed to localize the TJs.Enzyme-linked immunosorbent assay(ELISA)and RT-PCR were utilized to observe changes in interleukin(IL)-1βlevels.To investigate the role of NF-κB signaling activation and IL^(-1)βin Sema7A’s anti-barrier mechanism,we employed 0.1μmol/L IκB kinase 2(IKK2)inhibitor IV or 500 ng/mL IL^(-1)receptor(IL-1R)antagonist.●RESULTS:Treatment with Sema7A resulted in decreased TEER and increased permeability of Dextran-FITC in HCEs through down-regulating mRNA and protein levels of TJs in a time-and dose-dependent manner,as well as altering the localization of TJs.Furthermore,Sema7A stimulated the activation of inhibitor of kappa B alpha(IκBα)and expression of IL-1β.The anti-barrier function of Sema7A was significantly suppressed by treatment with IKK2 inhibitor IV or IL-1R antagonists.●CONCLUSION:Sema7A disrupts barrier function through its influence on NF-κB-mediated expression of TJ proteins,as well as the expression of IL-1β.These findings suggest that Sema7A could be a potential therapeutic target for the diseases in corneal epithelium.展开更多
基金Supported by JIPMER intramural research grant,Indian Council of Medical Research (ICMR),New Delhi,India,No. 3/1/3 J.R.F.-2016/LS/HRD
文摘A growing body of evidence suggests that tight junction(TJ)proteins play a crucial role in the pathogenesis of various diseases,including gastrointestinal(GI)cancer and hepatocellular carcinoma(HCC).TJ proteins primarily maintain the epithelial and endothelial cells intact together through integral proteins however,recent reports suggest that they also regulate gene expression necessary for cell proliferation,angiogenesis,and metastasis through adapter proteins such as zonula occludens(ZO).ZO proteins are membrane-associated cytosolic scaffolding proteins that modulate cell proliferation by interacting with several transcription factors.Reduced ZO proteins in GI cancer and HCC are correlated with tumor development and poor prognosis.Pubmed has searched for using the keyword ZO and gastric cancer,ZO and cancer,and ZO and HCC for the last ten years to date.This review summarized the role of ZO proteins in cell proliferation and their expression in GI cancer and HCC.Furthermore,therapeutic interventions targeting ZO in GI and liver cancers are reviewed.
基金Supported by Foundation of Chinese Medicine in Zhejiang Province Science and Technology,No.Z0102B002
文摘AIM:To study the effect of salvianolate on tight junctions(TJs) and zonula occludens protein 1(ZO-1) in small intestinal mucosa of cirrhotic rats.METHODS:Cirrhosis was induced using carbon tetrachloride.Rats were randomly divided into the untreated group,low-dose salvianolate(12 mg/kg) treatment group,medium-dose salvianolate(24 mg/kg) treatment group,and high-dose salvianolate(48 mg/kg) treatment group,and were treated for 2 wk.Another 10 healthy rats served as the normal control group.Histological changes in liver tissue samples were observed under a light microscope.We evaluated morphologic indices of ileal mucosa including intestinal villi width and thickness of mucosa and intestinal wall using a pathological image analysis system.Ultrastructural changes in small intestinal mucosa were investigated in the five groups using transmission electron microscopy.The changes in ZO-1 expression,a tight junction protein,were analyzed by immunocytochemistry.The staining index was calculated as the product of the staining intensity score and the proportion of positive cells.RESULTS:In the untreated group,hepatocytes showed a disordered arrangement,fatty degeneration was extensive,swelling was obvious,and disorganized lobules were divided by collagen fibers in hepatic tissue,which were partly improved in the salvianolate treated groups.In the untreated group,abundant lymphocytes infiltrated the fibrous tissue with proliferation of bile ducts,and collagen fibers gradually decreased and damaged hepatic lobules were partly repaired following salvianolate treatment.Compared with the untreated group,no differences in intestinal villi width between the five groups were observed.The villi height as well as mucosa and intestinal wall thickness gradually thickened with salvianolate treatment and were significantly shorter in the untreated group compared with those in the salvianolate treatment groups and normal group(P < 0.01).The number of microvilli decreased and showed irregular lengths and arrangements in the untreated group.The intercellular space between epithelial cells was wider.The TJs were discontinuous,which indicated disruption in TJ morphology in the untreated group.In the treated groups,the microvilli in the intestinal epithelium were regular and the TJs were gradually integrated and distinct.The expression of ZO-1 decreased in the small intestine of the untreated cirrhotic rats.The high expression rate of ZO-1 in ileal mucosa in the untreated group was significantly lower than that in the medium-dose salvianolate group(21.43% vs 64.29%,χ 2 = 5.25,P < 0.05),high-dose salvianolate group(21.43% vs 76.92%,χ 2 = 8.315,P < 0.01) and normal group(21.43% vs 90%,χ 2 = 10.98,P < 0.01).CONCLUSION:Salvianolate improves liver histopathological changes,repairs intestinal mucosa and TJ structure,and enhances ZO-1 expression in the small intestinal mucosa in cirrhotic rats.
文摘AIM To investigate the interaction of Zot withmicrotubule.METHODS Zot affinity column was applied topurify Zot-binding protein（s）from crudeintestinal cell lysates.After incubation at roomtemperature,the column was washed and theproteins bound to the Zot affinity column wereeluted by step gradient with NaCl(0.3 mol·L<sup>-1</sup>-0.5mol·L<sup>-1</sup>).The fractions were subjected to6.0%-15.0%（w/v）gradient SDS-PAGE andthen transferred to PVDF membrane for N-terminal sequencing.Purified Zot and tauprotein were blotted by using anti-Zot or anti-tauantibodies.Finally,purified Zot was tested in anin vitro tubulin binding assay.RESULTS Fractions from Zot affinity columnyielded two protein bands with a Mr of 60 kU and45kU respectively.The N-terminal sequence ofthe 60 kU band resulted identical to β-tubulin.Zot also cross-reacts with anti-tau antibodies.Inthe in vitro tubulin binding assay,Zot co-precipitate with Mt,further suggesting that Zotpossesses tubulin-binding properties.CONCLUSION Taken together,these resultssuggest that Zot regulates the permeability ofintestinal tight junctions by binding tointracellular Mt,with the subsequent activationof the intracellular signaling leading to thepermeabilization of intercellular tight junctions.
基金Supported by a Research Grant from Beijing Administration of Traditional Chinese Medicine(No.JJ 2008-015)
文摘Objective：To determine the relationship of thick greasy tongue fur formation and permeability of vascular endothelial cells（ECs）with the protein expression of zonula occludens-1（ZO-1）.Methods：Sprague Dawley rats were randomly divided into a model group of severe acute pancreatitis（SAP）and a sham-operated （SO）group.The SAP rats were further divided into two subgroups on the basis of tongue-coating status：a thick greasy tongue fur group（SAP-TGF）and a normal tongue fur group（SAP-NF）.Six rats were chosen randomly from every group mentioned above for an Evans blue assay 5 days after model establishment.For the histomorphology analysis,the expressions of ZO-1 protein and mRNA were studied by hematoxylin-eosin （HE）staining,transmission electron microscope,Western blot,and Q-PCR using blood and tongue tissues, which were collected from 8 rats randomly chosen from each group.Results：The papillae density of the rat tongue surface and the caryocinesis frequency of the basal layer were significantly increased in the SAP-TGF group compared with the SO group（P0.05）.Evans blue levels in the tongue tissue of the SAP-TGF group were significantly higher than that of the SO and SAP-NF groups（P0.05）.Vascular ECs were wider and obviously fissured in the SAP-TGF group under transmission electron microscope observation.The protein and mRNA expression of ZO-1 in the SAP-TGF group were lower than those in the SAP-NF（P0.05）.Conclusions： Reproductive activity enhancement of glossal epithelial cells was one of the main characteristics of thick greasy tongue fur formation.An increase in vasopermeability was closely associated with thick greasy tongue fur formation.Tight junction structural variation of vascular ECs might play an important role in the pathological and physiological process of thick greasy tongue fur formation.
文摘目的研究血清紧密连接蛋白闭锁小带蛋白1(zonula occludens 1,ZO-1)、咬合蛋白(occludin)、闭合蛋白-1(claudin-1)联合急性生理学与慢性健康状况Ⅱ(acute physiology and chronic health evaluationⅡ,APACHEⅡ)评分对高甘油三酯血症型重症急性胰腺炎(hypertriglyceridemia severe acute pancreatitis,HTG-SAP)并发急性肾损伤(acute kidney injury,AKI)的预测价值。方法选择2021年1月~2024年3月期间收治的96例HTG-SAP患者进行回顾性研究,根据是否并发AKI分为AKI组(n=38)和非AKI组(n=58)。比较两组临床资料及血清ZO-1、occludin、claudin-1水平的差异,筛选差异有统计学意义的因素作为自变量进行logistic回归分析并构建预测模型,通过受试者工作特征曲线分析各个指标对HTG-SAP并发AKI的预测价值。结果AKI组的APACHEⅡ评分、全身炎症反应综合征评分、甘油三酯、血肌酐、血尿素氮水平高于非AKI组,血清ZO-1、occludin、claudin-1水平低于非AKI组,差异有统计学意义(P<0.05);logistic回归分析显示,APACHEⅡ评分(OR=1.781,95%CI:1.065~2.981)升高以及血清ZO-1(0.956,95%CI:0.918~0.995)、occludin(0.967,95%CI:0.941~0.995)、claudin-1(0.193,95%CI:0.049~0.752)水平降低是HTG-SAP患者并发AKI的危险因素(P<0.05)。APACHEⅡ评分及血清ZO-1、occludin、claudin-1单独及联合模型预测HTG-SAP患者并发AKI的曲线下面积分别为0.821(95%CI:0.740~0.902)、0.852(95%CI:0.777~0.927)、0.877(95%CI:0.807~0.947)、0.936(95%CI:0.891~0.981)、0.998(95%CI:0.993~1.000)。结论血清紧密连接蛋白ZO-1、occludin、claudin-1水平降低及APACHEⅡ评分增加是HTG-SAP患者并发AKI的危险因素,根据四项指标建立预测模型对HTG-SAP患者并发AKI具有较好的预测价值。
基金Supported by the National Natural Science Foundation of China(No.81770889)Zhuhai Science and Technology Program(No.ZH22036201210134PWC).
文摘●AIM:To evaluate the role of semaphorin 7A(Sema7A)and its associated regulatory mechanisms in modulating the barrier function of cultured human corneal epithelial cells(HCEs).●METHODS:Barrier models of HCEs were treated with recombinant human Sema7A at concentrations of 0,125,250,or 500 ng/mL for 24,48,or 72h in vitro.Transepithelial electrical resistance(TEER)as well as Dextran-fluorescein isothiocyanate(FITC)permeability assays were conducted to assess barrier function.To quantify tight junctions(TJs)such as occludin and zonula occludens-1(ZO-1)at the mRNA level,reverse transcriptionpolymerase chain reaction(RT-PCR)analysis was performed.Immunoblotting was used to examine the activity of the nuclear factor-kappa B(NF-κB)signaling pathway and the production of TJs proteins.Immunofluorescence analyses were employed to localize the TJs.Enzyme-linked immunosorbent assay(ELISA)and RT-PCR were utilized to observe changes in interleukin(IL)-1βlevels.To investigate the role of NF-κB signaling activation and IL^(-1)βin Sema7A’s anti-barrier mechanism,we employed 0.1μmol/L IκB kinase 2(IKK2)inhibitor IV or 500 ng/mL IL^(-1)receptor(IL-1R)antagonist.●RESULTS:Treatment with Sema7A resulted in decreased TEER and increased permeability of Dextran-FITC in HCEs through down-regulating mRNA and protein levels of TJs in a time-and dose-dependent manner,as well as altering the localization of TJs.Furthermore,Sema7A stimulated the activation of inhibitor of kappa B alpha(IκBα)and expression of IL-1β.The anti-barrier function of Sema7A was significantly suppressed by treatment with IKK2 inhibitor IV or IL-1R antagonists.●CONCLUSION:Sema7A disrupts barrier function through its influence on NF-κB-mediated expression of TJ proteins,as well as the expression of IL-1β.These findings suggest that Sema7A could be a potential therapeutic target for the diseases in corneal epithelium.
