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Recombinant fibronectin polypeptide antagonizes hepatic failure induced by endotoxin in mice

Acta Pharmacologica Sinica 2004年第6期25卷 81-86页

作者： Yong WU, Yuan-zhong CHEN2, Hui-fang HUANG, Ping CHEN Fujian Institute of Hematology, Union Hospital, Fujian Medical University, Fuzhou 350001,China Fujian Institute of Hematology Union Hospital Fujian Medical University Fuzhou 350001

AIM: To study the preventive effect of recombinant human fibronectin (rhFN) polypeptide on hepatic failure in- duced by endotoxin in mice. METHODS: A cDNA fragment coding for Ile1363-Tyr1725 of human FN was inserted into the PinPoint Xa-3 plasmid, and the constructed plasmid was transformed into E coli BL21 (DE3) cells, and then the expression of rhFN polypeptide in DE3 cells was identified through SDS-PAGE. The target protein from the supernatant of bacteria lysate was purified through biotin-affinity chromatography. The bioactivity of the purified rhFN polypeptide was determined with cell adhesiveactivity. The survival rate was observed in endotoxemia mice injected with rhFN polypeptide. The tissue damage in hepatocyte was detected using histology, ultrastructure, and DNA fragmentation assay. RESULTS: The expression of rhFN polypeptide reached approximately 20 % of the total cellular protein. The adhesion ability of rhFN polypeptide was concentration-dependent. The value of EC50 was 0.8 nmol/L. The survival rate of endotoxemia mice sensitized by D-galactosamine (D-GalN) was 60 % in rhFN polypeptide treated group, while that of endotoxemia mice sensitized by D-GalN was 20 % in the control group (P<0.01). Histopathology showed that less necrosis occurred on the hepatocyte of endotoxemia mice in- jected with rhFN polypeptide compared with saline control. Ultrastructure and DNA fragmentation assay showed that no apoptotic hepatocyte was observed in the liver of rhFN-treated endotoxemia mice. CONCLUSION: Re- combinant fibronectin polypeptide antagonizes hepatic failure induced by endotoxin in mice.

关键词： fibronectins recombinant fusion proteins lipopolysaccharides acetylgalactosamine

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Expression of extracellular matrix proteins and vimentin in testes of azoospermic man： an immunohistochemical and morphometric study

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Asian Journal of Andrology 2002年第1期4卷 55-60页

作者： GulkKH ErdoT DepartmentofBiostatistics DepartmentofUrology

AIM: To investigate the changes in the extracellular matrix protein expression and the morphology of seminiferous tubules in the testis of 88 azoospermic men. METHODS: The patients were of the following categories: (1) 22 cases of Sertoli-cell-only syndrome, (2) 20 cases of spermatogenic arrest, and (3) 46 cases with hypospermatogenesis. Testicular sections were immunohistochemically stained for fibronectin, vimentin, laminin and collagen type IV. The seminiferous tubular diameter and the connective matrix zone (CMZ, the acellular zone between the basement membrane [BM] and the peritubular cells) thickness were measured. Seminiferous tubules were typed according to the thickness of the connective matrix in the lamina propria. The predominant tubule type and the Johnsen and Silber scores were determined. RESULTS: The mean tubular diameter were 119 +/- 27, 117 +/- 20, and 140 +/- 38 microm for Groups 1, 2, and 3, respectively. Both the laminin and the type IV collagen were localized to the epithelial BM and peritubular cells. In most of the tubules, BM and peritubular cells were separated by a homogenous acellular layer, the CMZ, in which laminin, type IV collagen, fibronectin and vimentin were not present. It is perceived that the worse the testicular histology, the higher the thickness of the CMZ. CONCLUSION: In testis with no or low sperm production, the diameter of the seminiferous tubules is decreased, the thickness of the seminiferous tubular wall is increased and a CMZ is formed between the peritubular cells and the BM. The thickness of CMZ is increasing with the advancement of testiclar deterioration. The most important morphologic predictive factor for spermiogenesis is the predominant

关键词： Adult Collagen Type IV Extracellular Matrix Proteins fibronectins Humans Immunohistochemistry Laminin Logistic Models Male Middle Aged Oligospermia Research Support Non-U.S. Gov't Seminiferous Epithelium Spermatogenesis Testis Vimentin

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Effectiveness of Qingre Lishi Yishen decoction on the glomerular fibrosis of immunoglobulin A nephropathy in a rat’s model

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Journal of Traditional Chinese Medicine 2019年第4期39卷 516-523页

作者： Dang Wanyu Hou Linyi Yan Huimin Wu Xiaoming Zhen Xiaofang Hao Jing Department of Traditional Chinese Medicine Beijing Children’s HospitalCapital Medical UniversityNational Center for Children’s healthCapital Medical UniversityNational Center for Children's healthBeijing 100045China

OBJECTIVE: To investigate the effect of the clinical effective prescription of Qingre Lishi Yishen decoction(QRLS) on the activation of mesangial cells in immunoglobulin A nephropathy(IgAN) ***: IgAN rat’s model was established by combine with intragastric administration of bovine serum albumin(BSA) + intravenous injection of lipopolysaccharide(LPS) by + subcutaneous injection of carbon tetrachloride(CCL4). Then the animals were randomly divided into four groups: control group, IgAN model group, IgAN model with Valsartan(Val) treatment group and IgAN model with QRLS treatment group. To observe the indexes of 24-h urine protein, renal function, deposition of immune complexes, expression of activation factor, fibrosis marker and inflammatory cytokines in four different ***: The Val or QRLS treatment group:(a) it reduced the immune complexes deposition of IgA in glomerular mesangial and inhibited mesangial cell proliferation;(b) it decreased the expression of smooth muscle actin(α-SMA), fibronectin(FN) and tumor necrosis factor alpha(TNF-α).CONCLUSION: The study suggested that QRLS ameliorate renal structure and function in IgAN rat’s model. Furthermore, we also observed that QRLS alleviated mesangial cells activation and matrix accumulation partly by decreasing the α-SMA,then to downregulated the expression of FN and TNF-α.

关键词： Glomerulonephritis,IGA Mesangial cells fibronectins Tumor necrosis factor-alpha Qingre Lishi Yishen decoction

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Role of precoating in artificial vessel endothelialization

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Chinese Journal of Traumatology 2004年第5期7卷 312-316页

作者：肖乐时德 Vascular Surgery Department First Affiliated Hospital of Chongqing University of Medical Sciences Chongqing 400016 China

As the progress of vascular surgery, artificial vessels have become the substitute for large and middle diameter vessels but have not for small diameter ones owing to thrombogenesis and occlusion within a short period of time after being applied. Artificial vessel endothelialization is one of the ideal methods to resolve such issue and has been improved continuously since Herring in 1978 put forward this term in the first time and utilized vascular endothelial cells (ECs) harvested from living animals to perform the test of artificial vessel endothelialization. However, human endothelial cells show little adhesion to the currently available vascular graft materials and some expanded polytetrafluoroethylene (ePTFE) grafts have shown only 10%+/-7% endothelial cell attachment rate (ECA, ie, attachment of ECs when incubated in vitro). Moreover, when the graft is exposed to pulsatile blood flow, a high proportion of cells are washed off from the lumen. Maximum cell loss occurs in the first 30-45 min after exposure to pulsatile flow, with up to 70% of cells lost. After that, a slower exponential loss occurs over the next 24 h. The lack of retention of cells could be partly overcome by sodding, but other techniques, involving engineering the lumen to improve ECA and endothelial cell retention rate (ECR, ie, retention of ECs when the grafts are exposed to pulsatile flow) have been developed. These include shear stress preconditioning, electrostatic charging and, above all, most successfully to date, precoating with EC specific adhesive glues that are mostly found in the extracellular basement membrane of blood vessels. The commonest are chemical coatings, preclotting, chemical bonding, and surface modifications.

关键词： Blood Vessel Prosthesis Coated Materials, Biocompatible Blood Vessel Prosthesis Implantation Cell Adhesion Collagen Comparative Study Female fibronectins Humans Laminin Male Materials Testing Prognosis Prosthesis Design Pulsatile Flow Risk Assessment Stress, Mechanical Treatment Outcome

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Effects of Huang Qi Wu Wu Decoction on Plasma Proteins in 70 Cases of Chronic Pulmonary Heart Disease

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Journal of Traditional Chinese Medicine 2000年第4期20卷 254-257页

作者：车洪柱罗凯利刘煜 463 PLA Hospital Shenyang 110042Liaoning Province

　　Simple immune agar diffusion test was used to assay the contents of 12 plasma proteins in 70 cases of chronic pulmonary heart disease treated by Huang Qi Wu Wu Decoction (黄芪五物汤), with the other 70 cases who were not given Huang Qi Wu Wu Decoction as the control group. The total clinical effective rate in the treatment group was 90.0%, while that in the control group was 75.7%, with a statistically significant difference between the two groups (P<0.05). In the treatment group, the levels of prealbumin, transferrin and fibronectin elevated obviously after treatment, and the contents of C-reactive protein, ceruloplasmin, haptoglobin, a 1-antitrypsin and a 1-acid glycoprotein decreased markedly (P<0.01). In the control group, only the levels of ceruloplasmin and C-reactive protein decreased significantly (P<0.05). It is shown that Huang Qi Wu Wu Decoction may enhance the therapeutic effects for pulmonary heart disease, regulate the metabolism of plasma proteins, and improve the life quality of the patients.

关键词： Aged C-Reactive Protein Chronic Disease Drugs, Chinese Herbal Female fibronectins Humans Male Middle Aged Pulmonary Heart Disease Transferrin

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Insulin-like growth factor binding protein-7 induces activation and transdifferentiation of hepatic stellate cells in vitro

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World Journal of Gastroenterology 2009年第26期15卷 3246-3253页

作者： Li-Xin Liu Shuai Huang Qian-Qian Zhang Yi Liu Dong-Mei Zhang Xiao-Hong Guo De-Wu Han Experimental Center of Science and Research of The First Teaching Hospital of Shanxi Medical University Taiyuan 030001 Shanxi Province China Institute of Liver Disease of Shanxi Medical University Taiyuan 030001 Shanxi Province China The First Teaching Hospital of Shanxi Medical University Taiyuan 030001 Shanxi Province China Key Laboratory of Cell Physiology Provincial Department of the Ministry of Education Taiyuan 030001 Shanxi ProvinceChina

AIM:To investigate the role of insulin-like growth factor binding protein-7 (IGFBP-7) in the activation and transdifferentiation of hepatic stellate cells (HSC) in ***:Rat HSC-T6 cells were cultured in separate dishes and treated with various concentration of transforming growth factor (TGF)-β1,IGFBP-7 or antiIGFBP-7 antibody for 24 *** supernatant or a cytoplasm suspension was obtained from cultured HSC,followed by transfer of cells to form cell-coated *** and Western blotting were used to analyze the expression of IGFBP-7 induced by TGF-β1 and the level of fibronectin,collagen and α-smooth muscle actin (SMA).The pro-apoptotic effect of antiIGFBP-7 antibody was determined by flow ***:Immunocytochemistry and Western blotting revealed that the expression of IGFBP-7 in TGF-β1 treated HSC was significantly up-regulated compared to that in the control *** addition,fibronectin,collagen and α-SMA also showed enhanced expression in accordance with the transdifferentiation process in a dose-dependent manner to some ***,flow cytometry suggested that anti-IGFBP-7 antibody induced apoptosis of activated HSC,which is responsible for the development of liver fibrosis,and may represent a novel pathway and target for therapeutic ***:IGFBP-7 showed increased expression in activated HSC and played an important role in the activation and transdifferentiation process of ***-7 antibody may ameliorate liver fibrogenesis.

关键词： Insulin-like growth factor-binding protein-7,Smooth muscle actin fibronectins Collagen type Ⅰ Hepatic stellate cells

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Activated focal adhesion kinase involved in adhesion and migration of vascular smooth muscle cells stimulated by fibronectin

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Chinese Medical Journal 2002年第4期115卷 494-497页

作者：尹航汪丽蕙霍勇彭旭夏春芳唐朝枢北京大学第一医院心脏科北京100034

OBJECTIVE: To study the effects of focal adhesion kinase (FAK) phosphorylation on smooth muscle cells (SMCs) adhesion and migration stimulated by fibronectin. METHODS: Adhesion and migration of cultured SMCs were stimulated by different concentrations of fibronectin (FN), FAK and its phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. SMCs adhesion and migration were also measured by morphological enumeration and modified Boyden Chambers, respectively. RESULTS: FAK were expressed when SMCs adhesion and migration were successfully simulated by different concentrations of FN. FAK phosphorylation were detected only at 20 microg/ml FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid and FAK phosphorylation was inhibited substantially. The SMCs migration rate in the 5 - 60 microg/ml FN groups was reduced by 17.89% - 27.67%. Cell migration stimulated by FN at 10, 20, 40 and 60 microg/ml were reduced by 23.26%, 21.63%, 19.31% and 17.88%, respectively (P

关键词： Animals Cell Adhesion Cell Movement Cells, Cultured DNA, Antisense Dose-Response Relationship, Drug Enzyme Activation fibronectins Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Muscle, Smooth, Vascular Phosphorylation Protein-Tyrosine Kinase Rats Transfection

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Fibronectin Glomerulopathy Caused by the Y973C Mutation in Fibronectin: A Case Report and Literature Review

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Chinese Medical Sciences Journal 2018年第1期33卷 60-63页

作者：李超文煜冰李航李明喜李学旺李雪梅中国医学科学院北京协和医学院北京协和医院肾内科

Fibronectin glomerulopathy is a rare autosomal dominant inherited glomerular disease associated with massive deposition of *** recently diagnosed fibronectin glomerulopathy in a 29-year-old woman presenting nephrotic *** analysis of fibronectin 1 gene showed heterozygosity for the Y973C ***,this mutation was not found in her *** had stable renal function but persistent nephrotic proteinuria after one-year follow-up.

关键词： fibronectins nephrotic syndrome mutation

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Animal study on expression of laminin and fibronectin in cornea during wound healing following alkali burn

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Chinese Journal of Traumatology 2003年第1期6卷 37-40页

作者：赵桂秋马轶群梁涛姜涛王传富张妍霞 DepartmentofOphthalmology

Objective: To observe the expression of laminin and fibronectin in alkali burned corneas in ***: A total of 18 normal Wistar rats were randomly divided into 6 groups (n=3 in each group). For each rat, one eye was injured by alkali burn, the other one was taken as the normal control. Then all the corneas were surgically removed and the expression of laminin and fibronectin was observed with immunohistochemistry respectively at 7 hours, 1 day, 3 days, 7 days, 14 days and 28 days after alkali burn. Results: Compared with that of the normal controls, the expression of laminin and fibronectin of the burned eyes was dramatically higher at 7 hours, reached peak at 14 days and decreased to the normal level at 28 days after alkali burn. Conclusions: In the process of wound healing after alkali burn, the expression of laminin and fibronectin increases dramatically, which suggests that laminin and fibronectin may participate in the process of corneal wound healing.

关键词： Alkaloids Cornea fibronectins Laminin

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