Remyelination failure is one of the main characteristics of multiple sclerosis and is potentially correlated with disease progression.Previous research has shown that the extracellular matrix is associated with remyel...Remyelination failure is one of the main characteristics of multiple sclerosis and is potentially correlated with disease progression.Previous research has shown that the extracellular matrix is associated with remyelination failure because remodeling of the matrix often fails in both chronic and progressive multiple sclerosis.Fibronectin aggregates are assembled and persistently exist in chronic multiple sclerosis,thus inhibiting remyelination.Although many advances have been made in the mechanisms and treatment of multiple sclerosis,it remains very difficult for drugs to reach pathological brain tissues;this is due to the complexity of brain structure and function,especially the existence of the blood-brain barrier.Therefore,herein,we review the effects of fibronectin aggregates on multiple sclerosis and the efficacy of different forms of drug delivery across the blood-brain barrier in the treatment of this disease.展开更多
The extracellular matrix surrounding oligodendrocytes plays an important role during myelination and remyelination in the brain.In many cases,the microenvironment surrounding demyelination lesions contains inhibitory ...The extracellular matrix surrounding oligodendrocytes plays an important role during myelination and remyelination in the brain.In many cases,the microenvironment surrounding demyelination lesions contains inhibitory molecules,which lead to repair failure.Accordingly,blocking the activity of these inhibitory factors in the extracellular matrix should lead to more successful remyelination.In the central nervous system,oligodendrocytes form the myelin sheath.We performed primary cell culture and found that a natural increase in fibronectin promoted the proliferation of oligodendrocyte progenitors during the initial stage of remyelination while inhibiting oligodendrocyte differentiation.Poly-L-ornithine blocked these inhibitory effects without compromising fibronectin’s pro-proliferation function.Experiments showed that poly-L-ornithine activated the Erk1/2 signaling pathway that is necessary in the early stages of differentiation,as well as PI3K signaling pathways that are needed in the mid-late stages.When poly-L-ornithine was tested in a lysolecithin-induced animal model of focal demyelination,it enhanced myelin regeneration and promoted motor function recovery.These findings suggest that poly-L-ornithine has the potential to be a treatment option for clinical myelin sheath injury.展开更多
Many studies have shown that fibronectin type III domain-containing protein 5(FDNC5) and brain-derived neurotrophic factor(BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an ...Many studies have shown that fibronectin type III domain-containing protein 5(FDNC5) and brain-derived neurotrophic factor(BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an environment that provides animals with multi-sensory stimulation and movement opportunities. An enriched environment has been shown to promote the regeneration of nerve cells, synapses, and blood vessels in the animal brain after cerebral ischemia;however, the exact mechanisms have not been clarified. This study aimed to determine whether an enriched environment could improve neurobehavioral functions after the experimental inducement of cerebral ischemia and whether neurobehavioral outcomes were associated with the expression of FDNC5 and BDNF. This study established ischemic mouse models using permanent middle cerebral artery occlusion(pMCAO) on the left side. On postoperative day 1, the mice were randomly assigned to either enriched environment or standard housing condition groups. Mice in the standard housing condition group were housed and fed under standard conditions. Mice in the enriched environment group were housed in a large cage, containing various toys, and fed with a standard diet. Sham-operated mice received the same procedure, but without artery occlusion, and were housed and fed under standard conditions. On postoperative days 7 and 14, a beam-walking test was used to assess coordination, balance, and spatial learning. On postoperative days 16–20, a Morris water maze test was used to assess spatial learning and memory. On postoperative day 15, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex were analyzed by western blot assay. The results showed that compared with the standard housing condition group, the motor balance and coordination functions(based on beam-walking test scores 7 and 14 days after operation), spatial learning abilities(based on the spatial learning scores from the Morris water maze test 16–19 days after operation), and memory abilities(based on the memory scores of the Morris water maze test 20 days after operation) of the enriched environment group improved significantly. In addition, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex increased in the enriched environment group compared with those in the standard housing condition group. Furthermore, the Pearson correlation coefficient showed that neurobehavioral functions were positively associated with the expression levels of FDNC5 and BDNF(r = 0.587 and r = 0.840, respectively). These findings suggest that an enriched environment upregulates FDNC5 protein expression in the ipsilateral cerebral cortex after cerebral ischemia, which then activates BDNF protein expression, improving neurological function. BDNF protein expression was positively correlated with improved neurological function. The experimental protocols were approved by the Institutional Animal Care and Use Committee of Fudan University, China(approval Nos. 20160858 A232, 20160860 A234) on February 24, 2016.展开更多
AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells.METHODS: ...AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells.METHODS: HTM cells were cultured to confluent and incubated with 0.4μmol/L Dex and/or 0.05μmol/L LAT-A.FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy.RESULTS: Dex up-regulated FN production in HTM cells,failed to do so when co-incubated with LAT-A.LAT-A decreased production of FN in cultured HTM cells.CONCLUSION: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma.It has an important prospect as an intraocular pressurelowering drug.展开更多
Objective: To observe the expression and significance of fibronectin and metalloproteinase-3(MMP-3) in patients with ankylosing spondylitis(AS). Methods: A total of 30 AS patients in our hospital and 30 healthy volunt...Objective: To observe the expression and significance of fibronectin and metalloproteinase-3(MMP-3) in patients with ankylosing spondylitis(AS). Methods: A total of 30 AS patients in our hospital and 30 healthy volunteers were selected in our study. Fibronectin and MMP-3 were measured and compared between these two groups. The AS group received sulfasalazine2 g daily for 3 months. Bath ankylosing spondylitis disease activity index, bath ankylosing spondylitis functional index, bath ankylosing spondylitis metrology index, erythrocyte sedimentation rate and C-reactive protein were compared before and after treatment.Pearson's linear-correlation analysis was used to determine relationships between parameters.Results: Totally 28 patients in the AS group completed the study. Fibronectin and MMP-3 in peripheral blood of AS patients were evidently higher than that in the normal control group(P<0.05). After treated by sulfasalazine, the level of expressing Fibronectin and MMP-3 significantly decreased compared with baseline values(P<0.05). Pearson's linear-correlation analysis showed that serum fibronectin and MMP-3 level had a positive correlation with bath ankylosing spondylitis disease activity index global assessment, spine pain, night pain, general pain, erythrocyte sedimentation rate and C-reactive protein(P<0.05). Conclusions: The expression of fibronectin and MMP-3 in AS patients were significantly higher than that in the normal control group, and they all decreased significantly after treatment. It indicated that both fibronectin and MMP-3 were correlated closely with the onset of AS.展开更多
The authors found experimentally that (1) fibronectin enhanced the healing of rabbit corneal epithelium after alkali burn and prevented the secondary breakdown; (2) it rapidly deposited on the denuded basement membran...The authors found experimentally that (1) fibronectin enhanced the healing of rabbit corneal epithelium after alkali burn and prevented the secondary breakdown; (2) it rapidly deposited on the denuded basement membrane to disappear as epithelial cells slided over, and (3) ultrastructurally, the neighbouring epithelial cells became flattened, with filopodia at the advancing edge, and extended to the wounded areas at 24 hours after the burn. However, the epithelial defects recurred 72 hours after the burn...展开更多
The adsorption behavior of heparin and fibronectin was studied by quartz crystal microbalance with dissipation(QCM-D), and the interaction between heparin and fibronectin was evaluated using immunochemistry and isothe...The adsorption behavior of heparin and fibronectin was studied by quartz crystal microbalance with dissipation(QCM-D), and the interaction between heparin and fibronectin was evaluated using immunochemistry and isothermal titration calorimetry(ITC) measurement. The results showed that there was competitive adsorption between heparin and fibronectin, and the preadsorption of fibronectin could prevent subsequent heparin adsorption to some extent, and the adsorbed Hep/Fn complex on the surface was in a rigid form. The bioactivity of heparin and fibronectin could be affected by the bulk concentration of each, and both heparin and fibronectin in Hep/Fn complex formed under p H 4 condition displayed larger bioactivity than that formed under p H 7 condition. Moreover, the fibronectin showed more exposed cell-binding sites at the p H value lower than physiological condition. The results of ITC further suggested that the interaction between heparin and fibronectin under p H 4 was stronger than under p H 7, and the complex was also more stable. The study brings forth the detailed interaction between heparin and fibronectin, which will be helpful for better understanding the interaction mechanism of the two biomolecules. The results may be potentially useful for the development of new generation of cardiovascular biomaterials.展开更多
Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690 -...Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690 -Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 in E. coli was very low, but that of CH63 was very high.The results suggests that Asp1961-Glu1978 in FN is a key sequence influencing the expression of triple-domain polypeptide in E. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography.Both of the cell-binding domains in the recombinant polypeptide were functional.The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function.展开更多
We prepared an anti-metastatic polypeptide, recombinant fibronectinpolypeptide CH50, and finished the preliminary identification of its functions. In this paper, we studied the effect of this polypeptide on the functi...We prepared an anti-metastatic polypeptide, recombinant fibronectinpolypeptide CH50, and finished the preliminary identification of its functions. In this paper, we studied the effect of this polypeptide on the function of macrophages. CH50 can significantly augment the production of nitric oxide(NO) by macrophages in a dose-dependent manner. The continuous presence of CH50 had a much stronger effect. In the presence of CH50, the cytotoxicity of macrophages to melanoma B16/F1 cells was significantly enhanced, and a stronger effect was obtained if CH50 was present continuously. CH50 polypeptide and IFN-r have a synergistic effect on the production of NO by macrophages and the cytotoxicity of macrophages on tumor cells. In the in vivo experiments, CH50 can inhibit the growth of tumor cells, and have a better effect in the presence of IFN-r. Our results suggest that recombinant fibronectin polypeptide CH50 has two functions: one is to inhibit the metastasis of tumor cells, and the other one is to augment the function of macrophages. And this polypeptide will be potentially useful in tumor therapy.展开更多
基金supported by the National Natural Science Foundation of China,Nos.82001282(to PW)and 81960232(to PW)Overseas Students’Innovation and Entrepreneurship Individual Project of Ningxia(2021)(to PW)+1 种基金Youth Talents Supporting Program of Ningxia Medical University and Ningxia,Nos.XT2019018(to PW),TJGC2019081(to PW)College Students’Innovation and En trepreneurship Training Program,No.X202210752038(to FYY)。
文摘Remyelination failure is one of the main characteristics of multiple sclerosis and is potentially correlated with disease progression.Previous research has shown that the extracellular matrix is associated with remyelination failure because remodeling of the matrix often fails in both chronic and progressive multiple sclerosis.Fibronectin aggregates are assembled and persistently exist in chronic multiple sclerosis,thus inhibiting remyelination.Although many advances have been made in the mechanisms and treatment of multiple sclerosis,it remains very difficult for drugs to reach pathological brain tissues;this is due to the complexity of brain structure and function,especially the existence of the blood-brain barrier.Therefore,herein,we review the effects of fibronectin aggregates on multiple sclerosis and the efficacy of different forms of drug delivery across the blood-brain barrier in the treatment of this disease.
基金supported by the National Nature Science Foundation of China,Nos.81371338(to HF)and 82071369(PPY)。
文摘The extracellular matrix surrounding oligodendrocytes plays an important role during myelination and remyelination in the brain.In many cases,the microenvironment surrounding demyelination lesions contains inhibitory molecules,which lead to repair failure.Accordingly,blocking the activity of these inhibitory factors in the extracellular matrix should lead to more successful remyelination.In the central nervous system,oligodendrocytes form the myelin sheath.We performed primary cell culture and found that a natural increase in fibronectin promoted the proliferation of oligodendrocyte progenitors during the initial stage of remyelination while inhibiting oligodendrocyte differentiation.Poly-L-ornithine blocked these inhibitory effects without compromising fibronectin’s pro-proliferation function.Experiments showed that poly-L-ornithine activated the Erk1/2 signaling pathway that is necessary in the early stages of differentiation,as well as PI3K signaling pathways that are needed in the mid-late stages.When poly-L-ornithine was tested in a lysolecithin-induced animal model of focal demyelination,it enhanced myelin regeneration and promoted motor function recovery.These findings suggest that poly-L-ornithine has the potential to be a treatment option for clinical myelin sheath injury.
基金supported by the National Natural Science Foundation of China,Nos.81601961(to KWY),81672242(to YW)the Key Construction Projects of Shanghai Health and Family Planning on Weak Discipline,China,No.2015ZB0401(to YW)
文摘Many studies have shown that fibronectin type III domain-containing protein 5(FDNC5) and brain-derived neurotrophic factor(BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an environment that provides animals with multi-sensory stimulation and movement opportunities. An enriched environment has been shown to promote the regeneration of nerve cells, synapses, and blood vessels in the animal brain after cerebral ischemia;however, the exact mechanisms have not been clarified. This study aimed to determine whether an enriched environment could improve neurobehavioral functions after the experimental inducement of cerebral ischemia and whether neurobehavioral outcomes were associated with the expression of FDNC5 and BDNF. This study established ischemic mouse models using permanent middle cerebral artery occlusion(pMCAO) on the left side. On postoperative day 1, the mice were randomly assigned to either enriched environment or standard housing condition groups. Mice in the standard housing condition group were housed and fed under standard conditions. Mice in the enriched environment group were housed in a large cage, containing various toys, and fed with a standard diet. Sham-operated mice received the same procedure, but without artery occlusion, and were housed and fed under standard conditions. On postoperative days 7 and 14, a beam-walking test was used to assess coordination, balance, and spatial learning. On postoperative days 16–20, a Morris water maze test was used to assess spatial learning and memory. On postoperative day 15, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex were analyzed by western blot assay. The results showed that compared with the standard housing condition group, the motor balance and coordination functions(based on beam-walking test scores 7 and 14 days after operation), spatial learning abilities(based on the spatial learning scores from the Morris water maze test 16–19 days after operation), and memory abilities(based on the memory scores of the Morris water maze test 20 days after operation) of the enriched environment group improved significantly. In addition, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex increased in the enriched environment group compared with those in the standard housing condition group. Furthermore, the Pearson correlation coefficient showed that neurobehavioral functions were positively associated with the expression levels of FDNC5 and BDNF(r = 0.587 and r = 0.840, respectively). These findings suggest that an enriched environment upregulates FDNC5 protein expression in the ipsilateral cerebral cortex after cerebral ischemia, which then activates BDNF protein expression, improving neurological function. BDNF protein expression was positively correlated with improved neurological function. The experimental protocols were approved by the Institutional Animal Care and Use Committee of Fudan University, China(approval Nos. 20160858 A232, 20160860 A234) on February 24, 2016.
基金National Natural Science Foundation of China (No.30772379)Education Department Project of Hunan Province, China(No.04A049)
文摘AIM: To determine the effects of a low dose latrunculin (LAT)-A on dexamethasone (Dex)-induced upregulation of extracellular matrix proteins fibronectin (FN) in cultured human trabecular meshwork (HTM) cells.METHODS: HTM cells were cultured to confluent and incubated with 0.4μmol/L Dex and/or 0.05μmol/L LAT-A.FN expression in HTM cells was evaluated by Western blot and immunofluorescence microscopy.RESULTS: Dex up-regulated FN production in HTM cells,failed to do so when co-incubated with LAT-A.LAT-A decreased production of FN in cultured HTM cells.CONCLUSION: This study indicated that LAT-A may modulate the expression of fibronectin in trabecular meshwork to achieve treatment for steroids and other types of glaucoma.It has an important prospect as an intraocular pressurelowering drug.
基金supported by Chinese Medicine Research Program of Zhejiang Province(No.2015ZB121)
文摘Objective: To observe the expression and significance of fibronectin and metalloproteinase-3(MMP-3) in patients with ankylosing spondylitis(AS). Methods: A total of 30 AS patients in our hospital and 30 healthy volunteers were selected in our study. Fibronectin and MMP-3 were measured and compared between these two groups. The AS group received sulfasalazine2 g daily for 3 months. Bath ankylosing spondylitis disease activity index, bath ankylosing spondylitis functional index, bath ankylosing spondylitis metrology index, erythrocyte sedimentation rate and C-reactive protein were compared before and after treatment.Pearson's linear-correlation analysis was used to determine relationships between parameters.Results: Totally 28 patients in the AS group completed the study. Fibronectin and MMP-3 in peripheral blood of AS patients were evidently higher than that in the normal control group(P<0.05). After treated by sulfasalazine, the level of expressing Fibronectin and MMP-3 significantly decreased compared with baseline values(P<0.05). Pearson's linear-correlation analysis showed that serum fibronectin and MMP-3 level had a positive correlation with bath ankylosing spondylitis disease activity index global assessment, spine pain, night pain, general pain, erythrocyte sedimentation rate and C-reactive protein(P<0.05). Conclusions: The expression of fibronectin and MMP-3 in AS patients were significantly higher than that in the normal control group, and they all decreased significantly after treatment. It indicated that both fibronectin and MMP-3 were correlated closely with the onset of AS.
文摘The authors found experimentally that (1) fibronectin enhanced the healing of rabbit corneal epithelium after alkali burn and prevented the secondary breakdown; (2) it rapidly deposited on the denuded basement membrane to disappear as epithelial cells slided over, and (3) ultrastructurally, the neighbouring epithelial cells became flattened, with filopodia at the advancing edge, and extended to the wounded areas at 24 hours after the burn. However, the epithelial defects recurred 72 hours after the burn...
基金Funded by the Natural Science Research Program of Jiangsu Education Department(No.13KJB310014)Natural Science Research Program of Jiangsu Province(No.BK20140429)
文摘The adsorption behavior of heparin and fibronectin was studied by quartz crystal microbalance with dissipation(QCM-D), and the interaction between heparin and fibronectin was evaluated using immunochemistry and isothermal titration calorimetry(ITC) measurement. The results showed that there was competitive adsorption between heparin and fibronectin, and the preadsorption of fibronectin could prevent subsequent heparin adsorption to some extent, and the adsorbed Hep/Fn complex on the surface was in a rigid form. The bioactivity of heparin and fibronectin could be affected by the bulk concentration of each, and both heparin and fibronectin in Hep/Fn complex formed under p H 4 condition displayed larger bioactivity than that formed under p H 7 condition. Moreover, the fibronectin showed more exposed cell-binding sites at the p H value lower than physiological condition. The results of ITC further suggested that the interaction between heparin and fibronectin under p H 4 was stronger than under p H 7, and the complex was also more stable. The study brings forth the detailed interaction between heparin and fibronectin, which will be helpful for better understanding the interaction mechanism of the two biomolecules. The results may be potentially useful for the development of new generation of cardiovascular biomaterials.
文摘Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690 -Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 in E. coli was very low, but that of CH63 was very high.The results suggests that Asp1961-Glu1978 in FN is a key sequence influencing the expression of triple-domain polypeptide in E. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography.Both of the cell-binding domains in the recombinant polypeptide were functional.The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function.
文摘We prepared an anti-metastatic polypeptide, recombinant fibronectinpolypeptide CH50, and finished the preliminary identification of its functions. In this paper, we studied the effect of this polypeptide on the function of macrophages. CH50 can significantly augment the production of nitric oxide(NO) by macrophages in a dose-dependent manner. The continuous presence of CH50 had a much stronger effect. In the presence of CH50, the cytotoxicity of macrophages to melanoma B16/F1 cells was significantly enhanced, and a stronger effect was obtained if CH50 was present continuously. CH50 polypeptide and IFN-r have a synergistic effect on the production of NO by macrophages and the cytotoxicity of macrophages on tumor cells. In the in vivo experiments, CH50 can inhibit the growth of tumor cells, and have a better effect in the presence of IFN-r. Our results suggest that recombinant fibronectin polypeptide CH50 has two functions: one is to inhibit the metastasis of tumor cells, and the other one is to augment the function of macrophages. And this polypeptide will be potentially useful in tumor therapy.
