OBJECTIVETo investigate the effects of spironolactone on the concentration of collagen type I, III in the myocardium of spontaneous hypertension rats (SHR).

METHODSTwenty 8-week male SHR were assigned randomly into spironolactone (SHR-SPIRO, n=10) and control groups (SHR-CON, n=10), sex-age matched Wistar Kyoto rats (WKY group, n=7) were also served as controls. The rats of SHR-SPIRO group were given 20 mg/(kg*d) of spironolactone, the rats of SHR-CON and WKY groups were given the same volume of distilled water. After 16 weeks, the concentration of collagen type I was analyzed with Western blot. The areas of collagen type I and III were observed under polarized light microscopy and the ratio of type I/III collagen was calculated through accumulation score.

RESULTSCompared with WKY group,the concentration of collagen type I in SHR-CON group was significantly higher (1.87 ±0.2 Compared with 1.21 ±0.7, P<0.05). After 16 weeks of treatment the concentration of collagen type I (1.42 ±0.05 Compared with 1.87 ±0.2, P<0.05) and I/III ratio in SHR-SPIRO group were significantly reduced (15.64 ±1.34 Compared with 20.8 ±3.04, P<0.05) compared with SHR-CON group; but there were no differences in accumulation area scores of collagen type III among three groups (368.3 ±30.2 Compared with 481.6 ±32.4 Compared with 406.2 ±45.3, P>0.05).

CONCLUSIONThe deposition of collagen type I in myocardium may be involved in myocardial fibrosis of SHR, and spironolactone can decrease the concentration of collagen type I, which may be one of the mechanisms for its therapeutic effects.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Male ; Mineralocorticoid Receptor Antagonists ; pharmacology ; Myocardium ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Spironolactone ; pharmacology

Although many different attempts have been made to reduce adhesions after tendon surgery, this complicated problem continues to be foremost among factors compromising tendon surgery and repair. The results of the finest tendon surgery and repair are frequently compromised by adhesions that restrict motion, decrease function and often lead to permanent deformation. Many substances and materials have been introduced into the area around primary flexor tendon repairs in an effort to prevent or diminish the adhesion formation. Various biochemical agents including antihistamines, anaboilic agents, lathyrogenic drug, betaamino-propionitrile and steroids, have been investigated and material such as nylon, cellophane, polyethylene film and silastic have been used to form pseudosheath. However the results with all of these method have been disappointing. Recently, interest has grown in a mucopolysaccharide found in synovial fluid hyaluronic acid. Synovial fluid normally contains a concentration of 2-3 mg/ml of hyaluronic acid. Preliminary investigations suggest that hyaluronic acid placed between the tendon and its sheath promote tendon healing and decrease adhesion formation. With this in mind, the present study was designed to examine the effect of hyaluronic acid on injured chicken flexor tendon healing and to determine the quantity and quality for adhesions in hyaluronic acid treated chicken compared to control. The stereomicroangiographic technique has enabled us to study the vascular process inside the tendon and surrounding tissues after injury of the tendon. For the study, the deep flexor tendon of the third toe of adult chicken with initial weight of about 2kg was used. Animals were divided into three groups and treated with different levels of hyaluronic acid. After 1,3,7,21,35 and 42 days postoperation, the animals were sacrificed and specimen were prepared. Obtained specimen were examined macroscopically and microscopically. At 1,3,5, and 7 weeks postoperation, microangiographic study were done and examind specimen by stereomicroscope. The results were as follows: 1. Though both the treated and control groups showed adhesion, hyaluronic acid treated tendon showed less adhesion tissue and better sliding properties as compared to the controls. 2. During the first week after operation, microangiographic studies showed failure of filling of all the vessels within the injured tendon. At 3 weeks highly vascular adhesions which extended throughout the injured site were observed. After then, the amount of vascularity decreased. 3. There were no difference microscopically between hyaluronic acid treated and control group at first week. After one week hyaluronic acid treated group showed less granulation tissue and less formation of collagen fibers. 4. There were no difference of healing process between hyaluronic acid treated and control group. From this results, it is suggested that hyaluronic acid is effective in reducing the adhesions after the tendon surgery.
Adult ; Animals ; Cellophane ; Chickens ; Collagen ; Granulation Tissue ; Histamine Antagonists ; Humans ; Hyaluronic Acid ; Methods ; Nylons ; Polyethylene ; Steroids ; Synovial Fluid ; Tendons ; Toes

BACKGROUNDRepeated attacks of bronchial asthma lead to different degrees of airway remodeling, the mechanism of which is not yet clear. Some evidences indicate that it is related to the excessive expression of some growth promotion factors. Angiotensin II is a polypeptide that may be involved in airway remodeling. To evaluate its role in airway remodeling in asthma, we observed the effects of an angiotensin II type 1 receptor antagonist (valsartan) on the expression of collagen III, collagen V, and transforming growth factor beta1 (TGF-beta1) mRNA and protein in the airway walls of sensitized rats.

METHODSForty Wistar rats were randomly divided into 5 groups: control group, sensitized group, and valsartan groups 1, 2, and 3. The rats in the sensitized group and in valsartan groups 1, 2, and 3 were sensitized and challenged with ovalbumin. Rats in control group were sensitized and challenged with 0.9% NaCl. Rats from valsartan groups 1, 2, and 3 were drenched with valsartan (10 microg, 20 microg, or 30 microg, respectively) at the time of the ovalbumin challenges. The expression of collagen III, collagen V, and TGF-beta1 protein were detected using immunohistochemical method in combination with image analysis methods. The expression of TGF-beta1 mRNA was detected by in situ hybridization.

RESULTSThe expression in the airways of collagen III and collagen V was significantly higher in rats from the sensitized group (7.73 +/- 0.81, 1.34 +/- 0.28) and from valsartan groups 1, 2, and 3 (5.73 +/- 0.64, 1.13 +/- 0.15; 4.96 +/- 0.51, 0.98 +/- 0.08; 4.43 +/- 0.35, 0.93 +/- 0.06, respectively) than those in the control group (2.65 +/- 0.38, 0.67 +/- 0.08, P < 0.05). In addition, collagen levels were significantly lower in valsartan groups 1, 2, and 3 than those from the sensitized group (P < 0.05). The expression of TGF-beta1 mRNA and protein in the airways was significantly higher in rats from the sensitized group (20.49% +/- 3.46%, 29.73% +/- 3.25%) and from valsartan groups 1, 2, and 3 (16.47% +/- 1.94%, 19.41% +/- 1.87%; 14.38% +/- 1.58%, 18.29% +/- 1.43%; 12.96% +/- 1.73%, 18.63% +/- 1.11%, respectively) than that from the control group (7.84% +/- 1.61%, 5.63% +/- 1.07%, P < 0.05). TGF-beta1 mRNA and protein levels were significantly lower in valsartan groups 1, 2, and 3 than that in the sensitized group (P < 0.05).

CONCLUSIONSAngiotensin II receptor antagonist valsartan can suppress synthesis of collagen III and collagen V by downregulating TGF-beta1 mRNA and protein expression. Valsartan can decrease airway remodeling and could play a role in asthma therapy.

Angiotensin Receptor Antagonists ; Animals ; Asthma ; physiopathology ; Bronchi ; metabolism ; Collagen Type III ; analysis ; Collagen Type V ; analysis ; Immunization ; Male ; Ovalbumin ; RNA, Messenger ; analysis ; Random Allocation ; Rats ; Rats, Wistar ; Tetrazoles ; pharmacology ; Transforming Growth Factor beta ; analysis ; Valine ; analogs & derivatives ; pharmacology ; Valsartan

Whether tranilast had antagonistic effect on proliferation inhibition and collagen synthesis promotion induced by TGF-beta2 in cultured human trabecular meshwork cells was investigated. Suspension of 1 x 10(4) cultured human trabecular meshwork cells of 3-5 passage was distributed in each well of a 96-well disk and divided into control group and experimental group. After 24 h, 0 microg/ml (control), 12.5 microg/ml, 25 microg/ml, 50 microg/ml tranilast with 3.2 ng/ml TGF-beta2 were added into the incubation medium. Another 24 h later, proliferation and collagen synthesis in cultured human trabecular meshwork cells were examined respectively by using tetrazolium-based semiautomated colormetric (MTT) assay and 3H-proline incorporation with liquid scintillation technique. The results showed absorbance (A) values of the experimental groups were 0.9036 +/- 0.3017, 1.1361 +/-0.1352, 1.2457 +/- 0.1524 according to the different concentrations of tranilast, and 0.8956 +/-0.1903 of the control group. In comparison with the control group, 25 microg/ml (q'= 3.23, P< 0.05), 50 microg/ml (q'=4.70, P<0.01) tranilast significantly antagonized the decrease of the A values induced by TGF-beta2 in the cultured human trabecular meshwork cells. In comparison with the control group [817.37+/-124.21 cpm/10(4) cells], 12.5 microg/ml (620.33+/-80.46 cpm/10(4) cells, q'= 4.26, P<0.05), 25 microg/ml (594.58+/-88.13 cpm/10(4) cells, q'=4.81, P<0.01), 50 microg/ml (418.64+/-67.90 cpm/10(4) cells, q'=8.62, P<0.01) tranilast significantly inhibited the incorporation of 3H-proline into the cultured human trabecular meshwork cells promoted by TGF-beta2 in a dose-dependent manner. It was concluded that tranilast had the antagonistic effect on the proliferation inhibition and collagen synthesis promotion induced by TGF-alpha2 in the cultured human trabecular meshwork cells.
Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Humans ; Trabecular Meshwork ; cytology ; metabolism ; Transforming Growth Factor beta ; antagonists & inhibitors ; Transforming Growth Factor beta2 ; ortho-Aminobenzoates ; pharmacology

OBJECTIVETo observe the effect of compound Puerarin on collagen IV of streptozotocin-induced diabetic rats.

METHODSDiabetic nephropathy rats were induced by intraperitoneal injection of streptozotocin (STZ). Rats were allocated randomly to control group (10), diabetes model group (10), Vitamin C group (10), Puerarin group (10), vitamin C plus Puerarin group (10). The study period lasted for 12 weeks. During and after the treatment, the general state, blood glucose levels, glycosylated hemoglobin, blood urea nitrogen, serum collagen IV, blood urea nitrogen, serum creatinine, urinary albumin excretion rate of the 24-hour, and clearance rate of creatinine collagen IV protein were determined by immunohistochemistoche analysis as well as type the gene expression of collagen IV alpha 1 mRNA were determined by in situ hybridization analysis in the kidney tissue of different groups.

RESULTS(1) Diabetes mellitus and renal function lesion occurred in the four groups. (2) Vitamin C and Puerarin could improve the general conditions of diabetic Rats, decrease blood urea nitrogen [(8.68 +/- 0.43), (7.98 +/- 0.47) and (5.76 +/- 0.82) micromol/L, serum creatinine [(74.68 +/- 8.20), (75.52 +/- 7.98) and (58.66 +/- 6.65) mmol/L], and urinary albumin excretion rate of the 24-hour [(18.40 +/- 0.37), (17.24 +/- 0.30) and (9.97 +/- 1.27) mg/24 h x 10(-3)]; increase clearance rate of creatinine [(0.59 +/- 0.21), (0.61 +/- 0.14) and (0.69 +/- 0.32) ml/min], the expression of collage IV absorbance [(111.56 +/- 14.61), (110.78 +/- 9.69) and (95.44 +/- 9.97) ] in the diabetic Rats were significantly inhibited at the same time.

CONCLUSIONThe compound Puerarin might have some functions on preventing ren by inhibiting expression of type IV collagen.

Animals ; Collagen Type IV ; antagonists & inhibitors ; biosynthesis ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetic Nephropathies ; drug therapy ; metabolism ; Isoflavones ; pharmacology ; therapeutic use ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

Acetaldehyde ; pharmacology ; Animals ; Anthracenes ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Collagen ; biosynthesis ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Flow Cytometry ; Hepatic Stellate Cells ; drug effects ; metabolism ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; Liver Cirrhosis, Alcoholic ; pathology ; prevention & control ; Rats ; Signal Transduction ; drug effects

OBJECTIVETo study the therapeutic effects to block the TGF-beta1 (transforming growth factor beta1) signal transduction by antisense Smad4 gene on experimental fibrotic liver.

METHODSUsing the rat model of liver fibrosis induced by Carbon Tetrachloride (CCl4)/ethanol, we transfected antisense Smad4 gene mediated by adenovirus via portal vein infusion into the liver, and observed the expression of Smad4 by Retro-Polymerase Chain Reaction (RT-PCR) and Western Blot. We also investigated the pathologic features and collagen expression.

RESULTSIn the non-therapeutic cirrhotic liver, the expression of Smad4 mRNA was significantly increased than normal liver, and so was the collagen I. After antisense Smad4 gene being transfected, the expression of Smad4 mRNA and that of collagen I in the therapeutic liver was significantly decreased, compared with the non-therapeutic cirrhotic liver. The fibrous degree of therapeutic liver was also reduced compared with the non-therapeutic fibrous liver.

CONCLUSIONThese results indicate that because antisense Smad4 gene could block TGF-beta1 signal transduction by reducing the expression of Smad4, so it could inhibit the production of extracellular matrix (ECM) and improve hepatic fibrosis.

Adenoviridae ; genetics ; Animals ; Antisense Elements (Genetics) ; therapeutic use ; Collagen Type I ; analysis ; DNA-Binding Proteins ; antagonists & inhibitors ; genetics ; Liver ; pathology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; therapy ; Male ; Rats ; Rats, Wistar ; Signal Transduction ; Smad4 Protein ; Trans-Activators ; antagonists & inhibitors ; genetics ; Transforming Growth Factor beta ; antagonists & inhibitors ; Transforming Growth Factor beta1

OBJECTIVETo study the effect of aldose reductase (AR) on expression of fibronectin and collagen IV in cultured rat renal mesangial cells (MsC).

METHODSAR expression plasmid vector (pCDNA3-AR) was constructed by restriction endonuclease digestion and ligation procedures. Stable expression of AR in MsC was established by Lipofectin transfection. Western blot and immunofluorescence analyses were performed to verify the transfection efficiency. Expression of fibronectin and collagen IV proteins were analyzed using Western blot.

RESULTSExpression of fibronectin and collagen IV in naive MsC treated with TGF-beta1 was upregulated in comparison to that of the untreated naive MsC (P < 0.01). MsC transfected with pCDNA3-AR showed a remarkable increase of expression of fibronectin and collagen IV (P < 0.01). Aldose reductase inhibitors (Sorbinil and Zopolrestat) significantly inhibited the expression of fibronectin and collagen IV in naive MsC (P < 0.05).

CONCLUSIONSOverexpression or inhibition of AR activity significantly alters the expression of fibronectin and collagen IV proteins in cultured rat MsC, suggesting that AR plays a significant role in the pathogenesis of glomerulosclersis.

Aldehyde Reductase ; antagonists & inhibitors ; genetics ; metabolism ; Animals ; Benzothiazoles ; pharmacology ; Cells, Cultured ; Collagen Type IV ; metabolism ; Fibronectins ; metabolism ; Genetic Vectors ; Imidazolidines ; pharmacology ; Mesangial Cells ; metabolism ; Phthalazines ; pharmacology ; Plasmids ; Rats ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo investigate the effect of rhein on the development of hepatic fibrosis.

METHODSThe animal models were made with carbon tetrachloride (CCl(4)) mixed with vegetable oil (3/2, v/v), which was injected subcutaneously twice a week for 6 weeks, and with 5% ethanol for free drinking water. At the same time, Rhein was administrated at the dose of 25 mg/kg or 100 mg/kg once a day for 6 weeks. The changes of both biochemical markers, such as the levels of alanine aminotransferase (ALT), hyaluronic acid (HA), procollagen type III (PCIII) in serum and SOD, malondialdehyde (MDA) in liver, and related histopathological parametres were determined.

RESULTSCompared with the model group, there were three kinds of changes in the larger quantity of rhein treated group. (1) The levels of ALT, HA, PCIII in serum and MDA in liver homogenate were decreased significantly (from 150 U/L +/- 16 U/L to 78 U/L +/- 18 U/L, 321 microg/L +/- 97 microg/L to 217 microg/L +/- 75 microg/L, 31 microg/L +/- 14 microg/L to 16 microg/L +/- 6 microg/L and 3.67 nmol/mg +/- 0.68 nmol/mg to 1.88 nmol/mg +/- 0.34 nmol/mg, respectively, t > or 2.977, P<0.01). However the level of SOD in liver was increased (from 62.45 NU/mg +/- 8.74 NU/mg to 91.26 NU/mg +/- 14.04 NU/mg, t=4.453, P<0.01). (2) The expressions of transforming growth factor beta 1 (TGF-beta 1) and alpha-smooth muscle actin (alpha-SMA) in liver were markedly reduced (P<0.05 and P<0.01). (3) The collagen staining positive area was decreased and the grade of fibrosis was reduced significantly in liver (P<0.05 and P<0.01).

CONCLUSIONRhein can protect hepatocyte from injury and prevent the progress of hepatic fibrosis in rats, which may associate with that rhein plays a role in antioxidation, anti-inflammation, inhibiting the expression of TGF-beta1 and suppressing the activation of hepatic stellate cells (HSCs).

Animals ; Anthraquinones ; pharmacology ; therapeutic use ; Anti-Inflammatory Agents ; pharmacology ; Antioxidants ; pharmacology ; Collagen ; analysis ; Liver ; drug effects ; pathology ; Liver Cirrhosis, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar ; Transforming Growth Factor beta ; antagonists & inhibitors ; Transforming Growth Factor beta1

OBJECTIVETo investigate the role of endothelin (ET) in the proliferation and collagen synthesis of human scar-derived fibroblasts and the modulation of its antagonists such as nitric oxide (NO), tetrandrine (Tet).

METHODSWith the cultured fibroblasts from the scarring tissue, the cell proliferation was determined by [3H]-TdR incorporation, while the collagen synthesis was evaluated by [3H]-proline incorporation.

RESULTSThe ET-1 was significantly increasing the proliferation and collagen synthesis of human scar-derived fibroblasts. The values of [3H]-TdR absorption in the 2.5 ng/ml, 25 ng/ml and 100 ng/ml of ET-1 groups were 1.8 times, 4 times and 4.9 times more than in the control group, respectively (P < 0.01), while the values of the [3H]-proline incorporation were 1.1 times, 3.1 times and 3.8 times respectively (P < 0.01). The fibroblasts, treated with 50 micrograms/ml of S-nitroso-N-acetyl penicillamine(SNAP), were no detectable effect on the basal level of DNA synthesis, but produced decreasing effect on the [3H]-TdR absorption (the rate of inhibition was 22.89%, P < 0.05). It was found that the SNAP inhibited the [3H]-proline incorporation in cultured fibroblasts, but the rate of [3H]-proline incorporation induced by ET-1 was unaltered. The Tet with 3 micrograms/ml, in which does not inhibit the basal level of DNA synthesis, was significantly decreasing the collagen synthesis and decreasing the ET-mediated DNA synthesis (the rate of inhibition was 33.21% (P < 0.01).

CONCLUSIONThese results indicate that the ET can obviously increase the proliferation and collagen synthesis of human scar-derived fibroblasts, but it can be partially antagonized by NO and Tet.

Benzylisoquinolines ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; pathology ; Collagen ; biosynthesis ; DNA ; biosynthesis ; Endothelins ; antagonists & inhibitors ; pharmacology ; Fibroblasts ; cytology ; radiation effects ; Humans ; Nitric Oxide ; metabolism ; pharmacology ; Proline ; metabolism ; S-Nitroso-N-Acetylpenicillamine ; pharmacology