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
目的观察腺苷预处理对大鼠脑缺血再灌注后基质金属蛋白酶9(MMP-9)和闭锁连接蛋白1(ZO-1)表达变化的影响。方法108只健康雄性SD大鼠随机分为对照组、模型组、腺苷组(n=36)。造模前腺苷组腹腔注射腺苷注射液,对照组和模型组大鼠在相同时...目的观察腺苷预处理对大鼠脑缺血再灌注后基质金属蛋白酶9(MMP-9)和闭锁连接蛋白1(ZO-1)表达变化的影响。方法108只健康雄性SD大鼠随机分为对照组、模型组、腺苷组(n=36)。造模前腺苷组腹腔注射腺苷注射液,对照组和模型组大鼠在相同时间点腹腔注射2 ml生理盐水。采用改良的线栓法栓塞右侧大脑中动脉阻断血流2 h后,将线拔出形成再灌注。再灌注24 h后,伊文思蓝(EB)渗透法测定血脑屏障通透性,检测MMP-9和ZO-1表达。结果模型组和腺苷组EB,脑含水量,MMP-9明显高于对照组,ZO-1明显低于对照组(P<0.01);模型组和腺苷组MMP-9/β肌动蛋白(β-actin)表达明显高于对照组(0.563±0.054和0.377±0.080 vs 0.242±0.021,P<0.01),ZO-1/β-actin表达明显低于对照组(0.186±0.042和0.393±0.075 vs 0.671±0.065,P<0.01)。腺苷组MMP-9/β-actin表达低于模型组,ZO-1/β-actin表达明显高于模型组(P<0.01)。结论腺苷预处理可以通过抑制MMP-9表达,增加ZO-1表达,改善血脑屏障完整性,减轻脑水肿与脑缺血再灌注后的神经损伤。展开更多
目的探讨急性创伤性硬膜外血肿(ATEDH)清除术患者紧密连接蛋白1(ZO1)、中性粒细胞/淋巴细胞比值(NLR)、血小板反应蛋白-1(TSP-1)动态变化及对预后的预测价值。方法选取2017年4月—2020年4月行ATEDH清除术169例,根据术后3个月预后情况分...目的探讨急性创伤性硬膜外血肿(ATEDH)清除术患者紧密连接蛋白1(ZO1)、中性粒细胞/淋巴细胞比值(NLR)、血小板反应蛋白-1(TSP-1)动态变化及对预后的预测价值。方法选取2017年4月—2020年4月行ATEDH清除术169例,根据术后3个月预后情况分为良好组147例、不良组22例,比较两组术前、术后3和7 d ZO1、NLR、TSP-1、格拉斯哥昏迷量表(GCS)评分、美国国立卫生研究院卒中量表(NIHSS)评分,采用Pearson相关分析ZO1、NLR、TSP-1与GCS、NIHSS评分的关系,采用Cox回归模型分析影响预后的相关因素,采用受试者工作特征(ROC)曲线分析预测预后的价值。结果与良好组比较,不良组术后3和7 d ZO1、NLR、TSP-1及NIHSS评分升高,GCS评分降低(P<0.01)。术后3和7 d ZO1、NLR、TSP-1与GCS评分呈负相关,与NIHSS评分呈正相关(P<0.05,P<0.01)。术后3和7 d ZO1、NLR、TSP-1均为影响ATEDH清除术患者预后的相关因素(P<0.01)。术后7 d,三者联合预测患者预后的曲线下面积最大。结论ZO1、NLR、TSP-1与ATEDH清除术患者意识状态、神经功能损伤程度及预后有关,联合检测术后7 d ZO1、NLR、TSP-1对患者预后具有良好预测价值。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
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
BACKGROUND Altered tight junction(TJ)proteins are correlated with carcinogenesis and tumor development.Nimbolide is a tetranotriterpenoid that has been shown to have antioxidant and anti-proliferative properties;howev...BACKGROUND Altered tight junction(TJ)proteins are correlated with carcinogenesis and tumor development.Nimbolide is a tetranotriterpenoid that has been shown to have antioxidant and anti-proliferative properties;however,its anticancer effects and molecular mechanism in hepatocellular carcinoma(HCC)remains obscure.AIM To investigate the effect of nimbolide on TJ proteins,cell cycle progression,and hepatic inflammation in a mouse model of HCC.METHODS HCC was induced in male Swiss albino mice(CD-1 strain)by a single intraperitoneal injection of 100 mg/kg diethylnitrosamine(DEN)followed by 80 ppm N-nitrosomorpholine(NMOR)in drinking water for 28 wk.After 28 wk,nimbolide(6 mg/kg)was given orally for four consecutive weeks in DEN/NMOR induced HCC mice.At the end of the 32nd week,all the mice were sacrificed and blood and liver samples were collected for various analyses.Macroscopic examinations of hepatic nodules were assessed.Liver histology and HCC tumor markers such as alpha-fetoprotein(AFP)and glypican-3 were measured.Expression of TJ proteins,cell proliferation,and cell cycle markers,inflammatory markers,and oxidative stress markers were analyzed.In silico analysis was performed to confirm the binding and modulatory effect of nimbolide on zonula occludens 1(ZO-1),nuclear factor of kappa light polypeptide gene enhancer in B-cells(NF-κB),and tumor necrosis factor alpha(TNF-α).RESULTS We found nimbolide treatment at a concentration of 6 mg/kg to HCC mice reduced hepatic tumor size by 52.08%and tumor volume(P<0.01),and delayed tumor growth in HCC mice with a concomitant reduction in tumor markers such as AFP levels(P<0.01)and glypican-3 expression(P<0.05).Furthermore,nimbolide treatment increased tight junction proteins such as ZO-1 and occludin expression(P<0.05,respectively)and reduced ZO-1 associated nucleic acid binding protein expression(P<0.001)in HCC mice liver.Nimbolide treatment to HCC mice also inhibited cell proliferation and suppressed cell cycle progression by attenuating proliferating cell nuclear antigen(P<0.01),cyclin dependent kinase(P<0.05),and CyclinD1(P<0.05)expression.In addition,nimbolide treatment to HCC mice ameliorated hepatic inflammation by reducing NF-κB,interleukin 1 beta and TNF-αexpression(P<0.05,respectively)and abrogated oxidative stress by attenuating 4-hydroxynonenal expression(P<0.01).Molecular docking studies further confirmed that nimbolide interacts with ZO-1,NF-κB,and TNF-α.CONCLUSION Our current study showed for the first time that nimbolide exhibits anticancer effect by reducing tumor size,tumor burden and by suppressing cell cycle progression in HCC mice.Furthermore,nimbolide treatment to HCC mice ameliorated inflammation and oxidative stress,and improved TJ proteins expression.Consequently,nimbolide could be potentially used as a natural therapeutic agent for HCC treatment,however further human studies are warranted.展开更多
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 elucidate the role of vascular endothelial growth factor-165b(VEGF-165b)in blood-retinal barrier(BRB)injury in the rat acute glaucoma model.METHODS:In this study,the rat acute high intraocular pressure(HIOP)mod...AIM:To elucidate the role of vascular endothelial growth factor-165b(VEGF-165b)in blood-retinal barrier(BRB)injury in the rat acute glaucoma model.METHODS:In this study,the rat acute high intraocular pressure(HIOP)model was established before and after intravitreous injection of anti-VEGF-165b antibody.The expression of VEGF-165b and zonula occludens-1(ZO-1)in rat retina was detected by double immunofluorescence staining and Western blotting,and the breakdown of BRB was detected by Evans blue(EB)dye.RESULTS:The intact retina of rats expressed VEGF-165b and ZO-1 protein,which were mainly located in the retinal ganglion cell layer and the inner nuclear layer and were both co-expressed with vascular endothelial cell markers CD31.After acute HIOP,the expression of VEGF-165b was up-regulated;the expression of ZO-1 was down-regulated at 12h and then recovered at 3d;EB leakage increased,peaking at 12h.After intravitreous injection of anti-VEGF-165b antibody,the expression of VEGF-165b protein was no significantly changed;and the down-regulation of the expression of ZO-1 was more obvious;EB leakage became more serious,peaking at 3d.EB analysis also showed that EB leakage in the peripheral retina was greater than that in the central retina.CONCLUSION:The endogenous VEGF-165b protein may protect the BRB from acute HIOP by regulating the expression of ZO-1.The differential destruction of BRB after acute HIOP may be related to the selective loss of retinal ganglion cells.展开更多
目的探讨游泳运动对β淀粉样前体蛋白(APP)/早老素1(PS1)双转基因小鼠闭锁小带蛋白1(ZO-1)、闭合蛋白(occludin)表达和学习记忆的影响。方法将20只APP/PS1小鼠随机分为模型组和游泳组每组10只,对照组选取同窝阴性小鼠10只。游泳组采用...目的探讨游泳运动对β淀粉样前体蛋白(APP)/早老素1(PS1)双转基因小鼠闭锁小带蛋白1(ZO-1)、闭合蛋白(occludin)表达和学习记忆的影响。方法将20只APP/PS1小鼠随机分为模型组和游泳组每组10只,对照组选取同窝阴性小鼠10只。游泳组采用游泳运动干预,用新物体识别实验、免疫蛋白印迹法以及免疫荧光检测小鼠学习记忆能力、海马紧密连接蛋白ZO-1、occludin表达和海马CA1区神经元表达情况;硫黄素S染色法检测海马β淀粉样蛋白(Aβ)斑块沉积情况。结果与对照组比较,模型组辨别指数(RI)明显降低(P<0.01);与模型组比较,游泳组RI明显升高[(63.63±13.35)%vs(50.38±9.83)%,P<0.01]。与对照组比较,模型组ZO-1、occludin明显降低(P<0.05,P<0.01);与模型组比较,游泳组ZO-1、occludin明显增加(0.81±0.02 vs 0.38±0.08,1.07±0.03 vs 0.68±0.12,P<0.05)。与对照组比较,模型组神经元阳性表达明显减少(P<0.01);与模型组比较,游泳组神经元阳性表达明显增加,海马Aβ斑块沉积明显减少,差异有统计学意义(P<0.05)。结论游泳运动能够增加APP/PS1小鼠海马紧密连接蛋白ZO-1和occludin蛋白,减轻海马神经元损伤和降低海马Aβ沉积,改善APP/PS1小鼠学习记忆能力。展开更多
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.
文摘目的观察腺苷预处理对大鼠脑缺血再灌注后基质金属蛋白酶9(MMP-9)和闭锁连接蛋白1(ZO-1)表达变化的影响。方法108只健康雄性SD大鼠随机分为对照组、模型组、腺苷组(n=36)。造模前腺苷组腹腔注射腺苷注射液,对照组和模型组大鼠在相同时间点腹腔注射2 ml生理盐水。采用改良的线栓法栓塞右侧大脑中动脉阻断血流2 h后,将线拔出形成再灌注。再灌注24 h后,伊文思蓝(EB)渗透法测定血脑屏障通透性,检测MMP-9和ZO-1表达。结果模型组和腺苷组EB,脑含水量,MMP-9明显高于对照组,ZO-1明显低于对照组(P<0.01);模型组和腺苷组MMP-9/β肌动蛋白(β-actin)表达明显高于对照组(0.563±0.054和0.377±0.080 vs 0.242±0.021,P<0.01),ZO-1/β-actin表达明显低于对照组(0.186±0.042和0.393±0.075 vs 0.671±0.065,P<0.01)。腺苷组MMP-9/β-actin表达低于模型组,ZO-1/β-actin表达明显高于模型组(P<0.01)。结论腺苷预处理可以通过抑制MMP-9表达,增加ZO-1表达,改善血脑屏障完整性,减轻脑水肿与脑缺血再灌注后的神经损伤。
文摘目的探讨急性创伤性硬膜外血肿(ATEDH)清除术患者紧密连接蛋白1(ZO1)、中性粒细胞/淋巴细胞比值(NLR)、血小板反应蛋白-1(TSP-1)动态变化及对预后的预测价值。方法选取2017年4月—2020年4月行ATEDH清除术169例,根据术后3个月预后情况分为良好组147例、不良组22例,比较两组术前、术后3和7 d ZO1、NLR、TSP-1、格拉斯哥昏迷量表(GCS)评分、美国国立卫生研究院卒中量表(NIHSS)评分,采用Pearson相关分析ZO1、NLR、TSP-1与GCS、NIHSS评分的关系,采用Cox回归模型分析影响预后的相关因素,采用受试者工作特征(ROC)曲线分析预测预后的价值。结果与良好组比较,不良组术后3和7 d ZO1、NLR、TSP-1及NIHSS评分升高,GCS评分降低(P<0.01)。术后3和7 d ZO1、NLR、TSP-1与GCS评分呈负相关,与NIHSS评分呈正相关(P<0.05,P<0.01)。术后3和7 d ZO1、NLR、TSP-1均为影响ATEDH清除术患者预后的相关因素(P<0.01)。术后7 d,三者联合预测患者预后的曲线下面积最大。结论ZO1、NLR、TSP-1与ATEDH清除术患者意识状态、神经功能损伤程度及预后有关,联合检测术后7 d ZO1、NLR、TSP-1对患者预后具有良好预测价值。
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.
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.
基金Supported by JIPMER intramural research grantIndian Council of Medical Research(ICMR),New Delhi,India,No.3/1/3 J.R.F.-2016/LS/HRDDepartment of Biotechnology,Government of India,No.102/IFD/SAN/22/2013-14.
文摘BACKGROUND Altered tight junction(TJ)proteins are correlated with carcinogenesis and tumor development.Nimbolide is a tetranotriterpenoid that has been shown to have antioxidant and anti-proliferative properties;however,its anticancer effects and molecular mechanism in hepatocellular carcinoma(HCC)remains obscure.AIM To investigate the effect of nimbolide on TJ proteins,cell cycle progression,and hepatic inflammation in a mouse model of HCC.METHODS HCC was induced in male Swiss albino mice(CD-1 strain)by a single intraperitoneal injection of 100 mg/kg diethylnitrosamine(DEN)followed by 80 ppm N-nitrosomorpholine(NMOR)in drinking water for 28 wk.After 28 wk,nimbolide(6 mg/kg)was given orally for four consecutive weeks in DEN/NMOR induced HCC mice.At the end of the 32nd week,all the mice were sacrificed and blood and liver samples were collected for various analyses.Macroscopic examinations of hepatic nodules were assessed.Liver histology and HCC tumor markers such as alpha-fetoprotein(AFP)and glypican-3 were measured.Expression of TJ proteins,cell proliferation,and cell cycle markers,inflammatory markers,and oxidative stress markers were analyzed.In silico analysis was performed to confirm the binding and modulatory effect of nimbolide on zonula occludens 1(ZO-1),nuclear factor of kappa light polypeptide gene enhancer in B-cells(NF-κB),and tumor necrosis factor alpha(TNF-α).RESULTS We found nimbolide treatment at a concentration of 6 mg/kg to HCC mice reduced hepatic tumor size by 52.08%and tumor volume(P<0.01),and delayed tumor growth in HCC mice with a concomitant reduction in tumor markers such as AFP levels(P<0.01)and glypican-3 expression(P<0.05).Furthermore,nimbolide treatment increased tight junction proteins such as ZO-1 and occludin expression(P<0.05,respectively)and reduced ZO-1 associated nucleic acid binding protein expression(P<0.001)in HCC mice liver.Nimbolide treatment to HCC mice also inhibited cell proliferation and suppressed cell cycle progression by attenuating proliferating cell nuclear antigen(P<0.01),cyclin dependent kinase(P<0.05),and CyclinD1(P<0.05)expression.In addition,nimbolide treatment to HCC mice ameliorated hepatic inflammation by reducing NF-κB,interleukin 1 beta and TNF-αexpression(P<0.05,respectively)and abrogated oxidative stress by attenuating 4-hydroxynonenal expression(P<0.01).Molecular docking studies further confirmed that nimbolide interacts with ZO-1,NF-κB,and TNF-α.CONCLUSION Our current study showed for the first time that nimbolide exhibits anticancer effect by reducing tumor size,tumor burden and by suppressing cell cycle progression in HCC mice.Furthermore,nimbolide treatment to HCC mice ameliorated inflammation and oxidative stress,and improved TJ proteins expression.Consequently,nimbolide could be potentially used as a natural therapeutic agent for HCC treatment,however further human studies are warranted.
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 the National Natural Science Foundation of China(No.81660217)Youth Foundation of the First Affiliated Hospital of Hainan Medical University(No.HYYFYPY201922)。
文摘AIM:To elucidate the role of vascular endothelial growth factor-165b(VEGF-165b)in blood-retinal barrier(BRB)injury in the rat acute glaucoma model.METHODS:In this study,the rat acute high intraocular pressure(HIOP)model was established before and after intravitreous injection of anti-VEGF-165b antibody.The expression of VEGF-165b and zonula occludens-1(ZO-1)in rat retina was detected by double immunofluorescence staining and Western blotting,and the breakdown of BRB was detected by Evans blue(EB)dye.RESULTS:The intact retina of rats expressed VEGF-165b and ZO-1 protein,which were mainly located in the retinal ganglion cell layer and the inner nuclear layer and were both co-expressed with vascular endothelial cell markers CD31.After acute HIOP,the expression of VEGF-165b was up-regulated;the expression of ZO-1 was down-regulated at 12h and then recovered at 3d;EB leakage increased,peaking at 12h.After intravitreous injection of anti-VEGF-165b antibody,the expression of VEGF-165b protein was no significantly changed;and the down-regulation of the expression of ZO-1 was more obvious;EB leakage became more serious,peaking at 3d.EB analysis also showed that EB leakage in the peripheral retina was greater than that in the central retina.CONCLUSION:The endogenous VEGF-165b protein may protect the BRB from acute HIOP by regulating the expression of ZO-1.The differential destruction of BRB after acute HIOP may be related to the selective loss of retinal ganglion cells.
文摘目的探讨游泳运动对β淀粉样前体蛋白(APP)/早老素1(PS1)双转基因小鼠闭锁小带蛋白1(ZO-1)、闭合蛋白(occludin)表达和学习记忆的影响。方法将20只APP/PS1小鼠随机分为模型组和游泳组每组10只,对照组选取同窝阴性小鼠10只。游泳组采用游泳运动干预,用新物体识别实验、免疫蛋白印迹法以及免疫荧光检测小鼠学习记忆能力、海马紧密连接蛋白ZO-1、occludin表达和海马CA1区神经元表达情况;硫黄素S染色法检测海马β淀粉样蛋白(Aβ)斑块沉积情况。结果与对照组比较,模型组辨别指数(RI)明显降低(P<0.01);与模型组比较,游泳组RI明显升高[(63.63±13.35)%vs(50.38±9.83)%,P<0.01]。与对照组比较,模型组ZO-1、occludin明显降低(P<0.05,P<0.01);与模型组比较,游泳组ZO-1、occludin明显增加(0.81±0.02 vs 0.38±0.08,1.07±0.03 vs 0.68±0.12,P<0.05)。与对照组比较,模型组神经元阳性表达明显减少(P<0.01);与模型组比较,游泳组神经元阳性表达明显增加,海马Aβ斑块沉积明显减少,差异有统计学意义(P<0.05)。结论游泳运动能够增加APP/PS1小鼠海马紧密连接蛋白ZO-1和occludin蛋白,减轻海马神经元损伤和降低海马Aβ沉积,改善APP/PS1小鼠学习记忆能力。
