Experimental study of changes of TNF-α in rabbits with steroid-induced avascular necrosis of femoral head

INTRODUCTION Avascularnecrosisoffemoralhead(ANFH)isawell recognizedconditionofunknownaetiology wherebythecirculationofthebloodtofemoralheadis impaired.Eventuallytheinvolvedareaofbonediesand necrosisdevelops.Itmaybeidiopathicorassociated withsomediseases,suchastrauma,useofsteroids.ANFHinducedbysteroidisgettingmoreandmorewith theunreliableapplicationofsteroidinclinic.However,theexactmechanismofitsoccurrence,sofar,is uncertain.Theexperimenttriestoexploretheeffectof tumornecrosisfactorα(…     

Relationship of Tumor Necrosis Factor-α and Nitrogen Oxide with Treatment of Frequent Relapse Nephrotic Syndrome by Shenkangling(肾康灵)Granule in Children

Objective:To observe the relationship of tumor necrosis factor-α(TNF-α)and nitrogenoxide(NO)with the treatment of frequent relapse nephrotic syndrome(FRNS)and to explore the patho-genesis of FRNS and the therapeutic mechanism of Shenkangling(肾康灵,SKL)Granule in children.Methods:Sixty children suffering from FRFRNS were randomly divided into the treated group and controlgroup,30 in each,and the other 30 healthy children were taken as healthy group.The patients were trea-ted with prednisone for a long-term course,and those with no effect or partial effect shown were treatedwith additional Tripterygium or Cytoxan in the control group,while in the treated group patients weretreated with prednisone and additional SKL.The two groups were compared as to their changes of TNF-α,NO before and after treatment,and the relapses after treatment.Results:The levels of TNF-α and NO inthe sick children before treatment were markedly higher than those afer treatment and normal group(P<0.01).The positive correlation     

The changes of nitric oxide and hemorheology in steroid-induced avascular necrosis of femoral head

Objective： To explore the pathogenesis of avascular necrosis of femoral head （ANFH）, the early diagnosis index and the treatment effective index of ANFH in clinical practice. Methods： Twenty-four Japanese rabbits were divided into 2 groups： model group and control group. ANFH models were produced by intramuscular injection of large dosage of steroid to rabbits for 8 weeks. On the 4 th, 8 th week after the injection, two rabbits each time from each group were taken to observe the structure of femoral head by light microscope and scanning electron microscope. Four other stomach-empty rabbits from each group were also used to test the contents of Nitric Oxide （NO）, contents of the hemorheology indexes. Results： Compared with the control group, the rabbits in the model group exhibited osteoporosis of femoral head and more bone lacuna and more fat cells through light microscope. Through scanning electron microscope observation bone trabecula were broken and sunk, and collagen fibers on the surface of bone matrix became loosen and broken, more osteocyte had pyknosis, adipocyte in the medullary cavity were enlarged and subchondral arterioles and capillaries of the femoral head were pressed by adipocyte. Compared with the control group, the model rabbits contained less NO and obvious increase of the plasma viscosity （PV）, low blood viscosity （LBV）, erythrocyte hematocrit （HCT）, indices of erythrocyte rigidity （TK） and indices of erythrocyte aggregation （AI）, plasma fibrin level （PFL） （P〈0. 01） and an increase of erythrocyte electrophoresis rate （ERT） （P〈 0. 05）. High blood viscosity （HBV）, and erythrocyte sedimentation rate （ESR） were unchanged. Conclusion： The steroid-induced ANFH might be related to less NO and the abnormal hemorheology; and NO and hemorheology should be considered as an early diagnosis index for ANFH in clinical practice.     

Ginsenoside Rg1-induced alterations in gene expression in TNF-α stimulated endothelial cells

Background In China the ginseng root began to be used in medicine over 2000 years ago. Ginsenosides are the most important component isolated from ginseng. The authors investigated the effect of ginsenoside Rg1 on the spectrum of gene expression in the endothelial cells stimulated by TNF-α and further explored the potential molecular mechanism of endothelial protection by ginsenoside Rg1.
Methods Nitric oxide (NO) production in the cultured human umbilical vein endothelial cells(HUVECs) was measured by using an NO assay kit. A home-made oligonucleotide microarray containing approximately 400 cardiovascular disease-related genes was constructed. The alteration of the spectrum of gene expression induced by ginsenoside Rg1 in HUVECs which were activated by TNF-α were detected by oligonucleotide microarray analysis.
Results NO production in HUVECs was decreased significantly after TNF-α treatment, while pretreatment with ginsenoside Rg1 enhanced NO production in TNF-αstimulated HUVECs. Ginsenoside Rg1 affected the expression levels of genes involved in vascular constriction, cell adherence, coagulation, cell growth and signal transduction in TNF-αstimulated HUVECs.
Conclusions Ginsenoside Rg1 could enhance NO production and the expression of eNOS mRNA in TNF-α stimulated HUVECs. Ginsenoside Rg1 regulated sets of genes in endothelial cells and protected endothelial cells from TNF-αactivation. Microarray analysis provided us with valuable insights into the atheroprotective mechanism by gingsenoside Rg1.
     

Inhibitory Effect of Oxymatrine on Quartz-induced Secretion of TNF-α by the Pulmonary Alveolar Macrophages in the Fibroblast Proliferation

The massive proliferation of pulmonary fibroblasts is the common consequence shared by lung diseases. The over-expression of TNF-α plays an important role in the proliferation of pulmonary fibroblasts[1, 2]. Under patho-logical conditions, TNF-α is predominantly secreted by activated alveolar macrophages of lung. The high level of TNF-α acts on the receptors on the cell membrane of fibroblasts, mediates the conduction of intracellular sig-nals and eventually activates the nuclear factor…     

Effects of puerarin on plasma endothelin and serum tumor necrosis Factor-α in type 2 diabetes mellitus patients with vascular complications

Ｔｈｅｉｎｃｉｄｅｎｃｅｏｆｄｉａｂｅｔｅｓｍｅｌｌｉｔｕｓ（ＤＭ）ｈａｓｄｒａｓｔｉｃａｌｌｙｉｎｃｒｅａｓｅｄａｔｔｈｅｐｒｅｓｅｎｔｔｉｍｅｗｉｔｈｍｉ ｃｒｏｖａｓｃｕｌａｒａｎｄｌａｒｇｅｖａｓｃｕｌａｒｄｉｓｔｕｒｂａｎｃｅｓａｓｔｈｅｍｏｓｔ     

Clinical study on effect of Astragalus Injection (黄芪注射液) and its immuno-regulation action in treating chronic aplastic anemia

Objective:To observe the clinical effect of Astragalus Injection(黄芪注射液,AI) and its immuno-regulatory action in treating chronic aplastic anemia(CAA).Methods:Sixty patients with CAA were randomly assigned to two groups equally,both were treated with Stanozolol three times a day,2 mg each time through oral intake,but AI was given additionally to the patients in the treated group once a day via intravenous dripping.All were treated for 15 days as one therapeutic course and the whole medication lasted for more than 4 months totally,with follow-up adopted.The clinical effi cacy was estimated and the changes of T-lymphocyte subsets in peripheral blood as well as the serum levels of tumor necrosis factor-α(TNF-α) and interleukin-2(IL-2) were observed.Results:The total effective rate in the treated group was 83.3%(25/30),which was higher than that in the control group 66.7%(20/30),showing significant difference between them(P<0.05).Levels of hemoglobin,WBC,reticular cell and platelet were elevated in both groups after treatment,but the improvement was signif icantly better in the treated group than that in the control group with respect to the former three indexes(P<0.05).The level of CD4 increased and that of CD8 decreased signifi cantly after treatment in the treated group(P<0.05),which showed significant difference as compared with those in the control group(P<0.05).Levels of serum TNF-α and IL-2 lowered after treatment in both groups,but signifi cance only showed in the treated group(P<0.05).The degree of proliferation in bone marrow got raised signifi cantly and the percentage of non-hemopoietic cells reduced signifi cantly in the treated group after treatment,also showing significant difference to those in the control group(P<0.05).Conclusion:AI could promote the recovery of hemopoietic function,which might be through improving T-lymphocyte subsets and reducing the release of negative regulatory factors such as TNF-α and IL-2 to alleviate the inhibition on hemopoietic function.     

Gene expression and cellular localizations of tumor necrosis factor-α at the site of implanted bovine cancellous bone in mice

Cellularimmunityisanimportantimmunephenomenonintheregionofxenogeneicbonegraft.Tumornecrosisfactor--a(TNFQ)playsacriticallmmuno--modulatoryroleincellularprocessofrejection.RecentevidencesupportingtherelationshipbetweenTNFQandboneresorptlonhasbeenprovidedbytheworkofMingetal.LIJ.However,detectionofTNFamRNAatthesiteofimplantedbonehasnotyetbeenshownexperimentally.Inthisstudy,anonradloactiveinSitubybridizationtechniquewasselectedtolocalizeTNFaintheregionofimplantedxenogeneicboneandtoinvesti…     

Effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with IL-2 or TNF-α

Objective To study the effects of vitamin E on the proliferation and collagen synthesis of rat hepatic stellate cells treated with interleukin-2 (IL-2 ) or tumor necrosis factor-α (TNF-α). Methods Hepatic stellate cells were isolated from male Sprague-Dawley rats by using modified Friedman’s method. Using the isolated cells cultured and treated with IL-2 or TNF-α, we studied the effects of vitamin E on their proliferation and collagen synthesis through an (3)H-thymidine and (3)H-proline incorporation assay, as well as through observation of these cells under a contrary phase microscope. Results Adding IL-2 increased the both proliferation and collagen synthesis of hepatic stellate cells. Their proliferation was also increased by the addition of TNF-α, although it decreased collagen synthesis. Vitamin E had marked inhibitory effects on the ability of cells treated with IL-2 or TNF-α to reproduce or synthesize collagen. Conclusion Vitamin E can inhibit the proliferation and collagen synthesis of hepatic stellate cells. It is possible that vitamin E affects liver fibrosis through these activities.     

Therapeutic Effect of Propofol in the Treatment of Endotoxin-induced Shock in Rats

Lipopolysaccharide (LPS) ,amaincomponentofgram negativebacterialendotoxin[1] ,istheleadingcauseofsepsisorendotoxicshock ,andwhenadmin isteredexperimentallytoanimals ,mimicsthesameinflammatoryresponse The pathophysiologicalchangesseeninsepsisareoftennotduetotheinfec tiousorganismitselfbuttotheuncontrolledproduc tionofinflammatorymediators ,includingtumornecrosisfactor (TNF) α ,nitricoxide (NO)andneu trophilaccumulation Accumulationoftheseleadstotissuedamage ,ultimately producingthelethalit…     

Effects of shenmai injection on expression of TNF- α mRNA in peritoneal macrophages of scald mice

Objective To explore the effect of shenmai injection (SI) on expression of TNF- α mRNA in peritoneal macrophages (pMΦs) of scald mice. Methods BALB/c mice were inflicted with 11% of body surface area Ⅲ degree scald and injected intraperitoneally (ip) with SI daily for 5 days, and expression of TNF- α mRNA in pMΦs was determined by semi- quantitative RT- PCR.Results In scald mice, the expression of TNF- α mRNA in pMΦs increased significantly, but it was reduced obviously (P&lt;0.01) after SI administration, while the livability was increased markedly (P&lt;0.05). Conclusions For scald mice, the cause of death at early stage might be related to the high expression of TNF- α mRNA in pMΦs and the use of SI can decrease the death rate.     

Iliac periosteal graft with vascular pedicle in the treatment of avascular necrosis of the femoral head. Experimental study and clinical application.

From March 1983 to June 1987, 6 patients with avascular necrosis of the femoral head were treated with vascular pedicled iliac periosteum graft. Follow-up for 3 to 7.5 years showed satisfactory results. The patients experienced no hip pain, could walk freely, and resumed work after the treatment. X-ray films showed that the original defect of the femoral head was filled up and its outline was clear and smooth. The bone density of the femoral head and neck normalized. The experimental study indicated that the vascularized periosteum improves the blood supply and initiates the revascularization and osteogenesis of the femoral head. The authors conclude that the technique is suitable for the treatment of avascular necrosis of the femoral head of Stage II and III.

     

Dynamic Expression of Tumor Necrosis Factor-α and Vascular Endothelial Growth Factor in Rat Model of Pulmonary Emphysema Induced by Smoke Exposure

In order to explore the roles of tumor necrosis factor-α(TNF-α) and vascular endothelial growth factor(VEGF) in the pathogenesis of pulmonary emphysema,male Wistar rats were randomized into group A1,group A2.5 and group A4,each with smoke exposure for 1 month,2.5 months or 4 months,respectively.Group B1,group B2.5 and group B4 were used as non smoking controls at corresponding time points.TNF-α in bronchoalveolar lavage fluid(BALF) and expression of VEGF in lung tissue was determined by ELISA or by SABC immunohistochemistry assay either.Lung slices were stained with hematoxylin and eosin(HE).Results showed that in animal with smoke exposure the mean linear interceptor(Lm),an index of pulmonary emphysema and the content of TNF-α in BALF increased gradually,on contrary,the expression of VEGF in lung tissue decreased(P<0.05).This phenomenon was not obvious in animals without smoke exposure.Lm was negatively correlated to the VEGF expression(γ=-0.81,P<0.01) and positively correlated to TNF-α concentration(γ = 0.52,P<0.004),which implies that smoke exposure decreased the expression of VEGF and increased the expression of TNF-α.It is plausible to speculate that the imbalance of TNF-α and VEGF may play an important role in the pathogenesis of smoke-induced pulmonary emphysema.     

Relationship between Single Nucleotide Polymorphism in TNF-α Gene Promoter Region and Inhibitory Effects of Triptolide on TNF-α Production in Peripheral Blood Mononuclear Cells of Healthy Humans

Tumor necrosis factorα(TNF-α)is ani mpor-tant cytokine that has extensive biological activity.It plays key rolesinthe pathogenesis of rheumatoidarthritis(RA).The TNF-αlocus has yielded a va-riety of polymorphic sites such as-308,-238, 498andso on[1].Amongthese polymorphic sites,ithas been demonstrated that TNF-α-308single nucle-otide polymorphism(SNP)is closely related withthe susceptibility,response to drugs and outcomeof RA[2-5].Triptolide that is a main active compo-nent of Tript…     

Effect of Salvia Miltiorrhiza Bge on Left Ventricular Hypertrophy and the Expression of Tumor Necrosis Factor-α in Spontaneously Hypertensive Rats

The effects of salvia miltiorrhiza Bge (SMB) on left ventricular hypertrophy (LVH) and the expression of tumor necrosis factor-α (TNF-α) in the left ventricle of spontaneously hypertensive rats and the action mechanism were investigated. Normal Wistar-kyoto (WKY) rats were used as negative control, and spontaneously hypertensive rats (SHR) were randomly assigned to receive pla- cebo or SMB. SMB (1 g/kg·d) was injected intraperitoneally for 12 weeks. Systolic blood pressure (SBP) and left ventricular mass index (LVMI) were measured. HE, VG and immunohistochemical staining combined with computed morphometry were employed to evaluate the cardiomyocyte size, diameter, the collagen volume fraction (CVF), perivascular circumferential area (PVCA), and tumor necrosis factor-α (TNF-α) expression in the left ventricular tissue. The results showed, as compared with WKY rats, the SBP, LVMI, cardiomyocyte size, diameter, CVF, PCVA, and TNF-α expression were increased markedly in the 20-week-old spontaneously hypertensive rats. SMB decreased LVMI (P<0.01), size of cardiomyocytes (P<0.01), collagen volume fraction (P<0.01), perivascular circum- ferential area (P<0.01), and TNF-α expression (P<0.01), but had no effect on SBP (P>0.05). It was suggested that chronic administration of SMB could inhibit and reverse the development of LVH in spontaneously hypertensive rats independent of BP. TNF-α may be involved in the reversal mecha- nism of LVH by SMB.     

TNF-alpha-induced metastasis gene changes in MCF-7 cells

Objective: Studies have shown that TNF-αsecreted by tumor cells and macrophages infiltrated into the tumor microenvironment might promote the metastasis of a variety of malignant cancers, including breast cancer. The present study was designed to detect global metastasis-related gene expression changes of MCF-7 cells treated by low dose TNF-α and to further explore the mechanisms by which TNF-αcontributes to metastasis. Methods: MCF-7 cells were cultured and treated with low dose TNF-α(20 ng/ml). cDNA array analysis was applied to detect the metastasis related gene expressions. Results: A total of 36 gene expressions were significantly regulated by TNF-α. Functional analysis indicates that the altered genes belong to different functional group. Most of the genes changed may promote the metastasis of MCF-7 cells while the others may inhibit metastasis. The changes observed in gene expression following TNF-αwere somewhat time dependent. Conclusion: TNF-αcan enhance the invasive ability of MCF-7 cells, partly by regulating a series of metastasis related genes, and these genes may take part in every step of metastasis. Some of the genes deserve further study.     

Effects of Huogu I Formula (活骨I方) on correlated factors of bone regeneration in chickens with steroid-induced necrosis of femoral head

Objective:To study the mechanism of HuoguⅠFormula（活骨Ⅰ方） in treating osteonecrosis of femoral head.Methods:Forty-eight healthy female Leghorn chickens were randomly divided into control group,model group and HuoguⅠgroup,and each group consisted of 16 chickens.At the meantime of model establishment,chickens of the HuoguⅠgroup were administrated with decoction,while the model and control group with distilled water by gavage.At the 8th and 16th week after medication,blood samples were obtained for blood lipid detection while both sides of femoral head were harvested for the rest of examinations.Specifically, expressions of bone morphogenetic protein-2（BMP2）,transforming growth factor betal（TGFβ₁）,Smad4 and Smad7 were evaluated by immunohistochemistry,while expression of osteoprotegerin/receptor activator of nuclear factor kappaB ligand（OPG/RANKL） mRNA was detected by in situ hybridization.Results:Compared with the control group,serum levels of total cholesterol（TC）,triglyceride（TG） and low-density lipoprotein cholesterol （LDL-C） in the model group rose significantly.Positive cell counting of BMP2,TGFβ₁,Smad4 and OPG in femoral head of the model group dropped prominently.Positive cell counting of Smad7 and RANKL increased dramatically. In contrast with the model group,levels of TC,TG and LDL-C in Huogu I group reduced significantly.Positive cell counting of BMP2,TGFβ₁,Smad4 and OPG in femoral head of the Huogu I group increased prominently. Indices of Smad7 and RANKL both decreased significantly.Especially at the 8th week,these variations were more significant.Conclusion:HuoguⅠFormula is effective in promoting repair of necrotic femoral head by regulating the expressions of BMP2,TGFβ₁,Smads and OPG/RANKL of osteoclast in femoral head.