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OBJECTIVE To investigate the inhibitory effect and the possible mechanism of hydrogen sulfide （H2S） on the proliferation of cardiac fibroblasts. METHODS The heart of neonatal SD rats was collected， and cardiac fibroblasts were separated with differential centrifugation. Using sodium hydrosulfide as the donor of H2S， the effects of H2S on the proliferation of cardiac fibroblasts induced by angiotensin Ⅱ （Ang Ⅱ）， hydroxyproline content and the expression of sirtuin 3 （SIRT3） protein were detected. After SIRT3 knockdown with siRNA technology， the effects of H2S on the proliferation of cardiac fibroblasts induced by Ang Ⅱ， hydroxyproline content， the expressions of collagen Ⅰ （Col Ⅰ）， collagen Ⅲ （Col Ⅲ ） and optic atrophy protein 1 （OPA1） were detected. RESULTS H2S could inhibit the proliferation of Ang Ⅱ-induced cardiac fibroblasts， reduce the content of hydroxyproline and increase the expression of SIRT3 （P＜0.05）. After down-regulating the expression of SIRT3 with siRNA technology， the inhibition of H2S on the proliferation of Ang Ⅱ-induced cardiac fibroblasts and the reduction of hydroxyproline content were both inhibited， and the effect of H2S on reducing the expression of Col Ⅰ and Col Ⅲ and enhancing the expression of OPA1 was also significantly weakened. CONCLUSIONS H2S inhibits the proliferation of Ang Ⅱ -induced cardiac fibroblasts through increasing the expression of SIRT3.
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Objective To study the protective effect of Rhein on the kidney of type 2 diabetic rats induced by high fat diet.Methods A rat model of type 2 diabetes was induced by high fat diet combined with low dose streptozotocin 35 mg/kg.The diabetic rats were randomly divided into diabetes group, Low, middle and high rhein dose groups (50,100,150 mg/kg), metformin group (300 mg/kg) and normal control group.Blood glucose and urine micro albumin were measured at 0, 2, 4 and 8 weeks respectively.Serum creatinine, urea nitrogen, total cholesterol and triglyceride were measured at 8 weeks.HE staining was used to observe the pathological changes of renal tissue.Effects of rhein on the expression of transforming growth factor-β1 and Smad3 protein in renal tissue of diabetic rats were detected with Western Blot.Results The blood glucose and urine micro albumin in model group were significantly higher than those in normal control group.Each rhein dose group exhibited reduced blood glucose and urinary micro albumin in diabetic rats.The high rhein dose group showed significant reduction of blood glucose and urine micro albumin in diabetic rats (P<0.05).Compared with model group, rhein reduced the serum Cr, BUN, TC and TG values in each dose group, most significantly in the high rhein dose group (P<0.05).The obvious pathological changes of renal tissue in model group were observed with most improved changes in the high rhein dose group.The expression of TGF-β1 and Smad3 protein in renal tissue of diabetic rats decreased significantly (P<0.05).Conclusion Rhein has preventive effect on diabetic nephropathy.The mechanism may relate to the improvement of renal function, regulation of blood lipid and down regulation of TGF-β1 and Smad3 protein expression in renal tissue.
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Aim To discuss the effects and mechanism of Ganoderma lucidum polysaccharides and metformin on myocardial structure and hemodynamics in type 2 diabetic rats.Methods High fat diet combined with intraperitoneal injection of low dose streptozotocin 30 mg· kg -1 was applied to establish rat model of type 2 diabetes mellitus .The diabetic rats were randomly into normal control group ,diabetes group , ganoderma lucid-um polysaccharides group (600 mg· kg -1 ) , metformin group ( 600 mg · kg -1 ) , combination group ( ganoder-ma lucidum polysaccharides 300 mg · kg -1 +metform-in 300 mg· kg -1 ) .After 12 weeks′treatment,the lev-els of fasting serum glucose were determined and the hemodynamic parameters (LVSP,LVEDP,dp/dtmax,-dp/dtmax ) were determined.Collagen volume fraction ( CVF ) was detected by Van Gieson . Immunohisto-chemical method and Western blot were used to detect myocardial tissue MMP-2 protein expression .Results The fasting blood glucose was significantly decreased in the combined treatment group .Combined medication could significantly improve hemodynamic parameters in diabetic rats: reduced LVEP and raised LVEDP , dp/dtmax and -dp/dtmax .CVF was significantly decreased in combination group .The expression of MMP-2 in my-ocardial tissue was significantly inhibited .Conclusions The combination of Ganoderma lucidum polysaccha-ride and metformin can significantly improve the hemo-dynamic parameters in type 2 diabetic rats, and have a preventive effect on diabetic cardiomyopathy . The mechanism may be related to the down regulation of the expression of MMP-2.
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Aim To observe the effects of benazepril and irbesartan on myocardial collagen in chronic heart failure ( CHF ) rats and the possible mechanisms. Methods CHF model in rats was made by partially banding abdominal aorta to induce pressure overload cardiac hypertrophy. The rats were given benazepril or/and irbesartan for 8 weeks. Systolic blood pressure ( SBP) was monitored by rat tail artery. The ultrastruc-ture of myocardium was detected by transmission elec-tron microscope. The content of myocardial hydroxyproline and amount of cross-linked collagen were measured by hydrolysis method. The expression of type Ⅰ and Ⅲ collagen protein in myocardium was investigated by immunohistochemistry. The expression of p-38 MAPK and p-p38 MAPK proteins was detected by Western blot. Results Compared with the model of CHF, there was no significant difference in SBP af-ter treatment. The content of hydroxyproline in myocar-dium, degree of cross-linked collagen, as well as ex-pression of type I collagen, type Ⅲ collagen and p-p38 MAPK proteins were significantly decreased after treatment with benazepril or irbesartan ( P <0. 05 ) , and the combined treatment of them had better effects. Conclusion There is a synergistic effect of attenua-ting the quantity and quality of myocardial collagen in CHF rats by combined use of benazepril and irbesar-tan, which may be related to the down regulation of p-p38MAPK protein expression.
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Objective To study the effect of ganoderma lucidum polysaccharides combined with metformin on oxidative stress of type 2 diabetic rats. Methods SD rats were fed with high fat diet for 4 weeks and injected with streptozotocin (30 mg·kg-1 ) to produce type 2 diabetic model. The diabetic rats were randomly divided into diabetes model group, ganoderma lucidum polysaccharides group (600 mg·kg-1 ), metformin group (600 mg·kg-1 ), combination group (ganoderma lucidum polysaccharides 300 mg·kg-1+ metformin 300 mg·kg-1 ), After 12 weeks of treatment, the level of fasting blood glucose was determined, and the activity of superoxide dismutase ( SOD), malondialdehyde ( MDA), catalase ( CAT), glutathione peroxidase (GSH-Px), total cholesterol (TC) and triglyceride (TG) were detected. Results The levels of fasting blood glucose in the treatment groups were significantly lower than that in the diabetes model group (P<0. 01). Furthermore, fasting blood glucose in the combination group was significantly lower than that in ganoderma lucidum polysaccharides group and metformin group (P<0. 01). Compared with diabetes model group, serum TC and TG in the treatment groups were significantly lower (P<0. 05, P<0. 01). Serum TC and TG were significantly lower in the combination group than in ganoderma lucidum polysaccharides group and metformin group (P<0. 05, P<0. 01). Compared with diabetes model group, serum SOD levels in the treatment groups were significantly higher (P<0. 01). Compared with ganoderma lucidum polysaccharides group and metformin group, serum SOD levels in the combination group was significantly higher (P<0. 05). Compared with diabetes group, serum MDA levels in the treatment groups were significantly lower (P<0. 01). Serum MDA in the combination group was significantly lower than that in ganoderma lucidum polysaccharides group and metformin group ( P<0. 05). Compared with diabetes model group, serum CAT and GSH-Px in the treatment groups were significantly higher (P<0. 05, P<0. 01). Serum CAT and GSH-Px in the combination group were significantly higher than those in ganoderma lucidum polysaccharides group and metformin group (P<0. 05). Conclusion Ganoderma lucidum polysaccharides combined with metformin could effectively inhibit oxidantion stress in type 2 diabetic rats. The effect was better than ganoderma lucidum polysaccharides or metformin used alone. The possible mechanism may be related to increased activity of SOD, CAT, GSH-Px in vivo and regulation of dyslipidemia.
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Aim To study the effects of ganoderma lu-cidum polysaccharides and metformin on myocardial fi-brosis of type 2 diabetic rats and its mechanism. Methods SD rats were fed with high fat diet for 4 weeks, and then were injected with streptozotocin (30mg·kg-1 ) to replicate type 2 diabetic model. The diabetic rats were randomized into normal control group,diabetes group, ganoderma lucidum polysaccha-rides group ( 600 mg · kg-1 ) , metformin group ( 600 mg·kg-1 ) , and combination group( ganoderma lucid-um polysaccharides 300 mg·kg-1 +metformin 300 mg ·kg-1 ) . After 12 weeks’ treatment,the levels of fast-ing serum glucose were determined and the extent of myocardial fibrosis was observed by Picro-sirius red staining. The contents of AGEs in serum were deter-mined by fluorescence spectrophotometer. The activities of CAT and GSH-Px in myocardium were detected. Im-munohistochemical method and Western blot were used to detect myocardial tissue AGEs and CTGF protein ex-pression. Results Combination group could repress patho-proceeding of myocardial fibrosis efficiently, im-prove the activity of CAT and GSH-Px in myocardium and lower the concentration of AGEs in serum, as well as reduce the expression of AGEs and CTGF in myo-cardium. Conclusions Ganoderma lucidum polysac-charides and metformin could prevent myocardial fibro-sis. The possible mechanism may be related to repress-ing oxidative stress of myocardium, lowering serum AGEs and down regulating AGEs and CTGF of myocar-dium.
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Aim To investigate the effects and mecha of ganoderma lucidum polysaccharides and metformin on pathological changes of thoracic aorta in diabetic ratsandthemechanisms.Methods SDratswerefed with high fat diet for 4 weeks and injected with strepto-zotocin ( 30 mg · kg-1 ) to replicate type 2 diabetic model. The diabetic rats were randomly into diabetes group, ganoderma lucidum polysaccharides group ( 600 mg·kg-1 ) ,metformin group(600 mg·kg-1 ) ,combi-nation group ( ganoderma lucidum polysaccharides 300 mg· kg-1 + metformin 300 mg · kg-1 ) and normal control group. After 12 weeksˊ treatment, the levels of fasting serum glucose, the activity of catalase(CAT), glutathione peroxidase ( GSH-Px ) , total cholesterol (TC)and triglyceride(TG) in serum were detected. Pathological changes of thoracic aorta were observed by HE staining. Immunohistochemy and Western blot were used to detect thoracic aorta VEGF protein expression. Results Combination group could lower fasting serum glucose and blood fat significantly, meanwhile the ac-tivity of CAT and GSH-Px in serum was improved. The expression of VEGF in thoracic aorta was repressed. The result of HE staining suggested that the lipid de-posits in aortic endothelium in combination group were lessthanthoseinthemodelgroup.Conclusions Ga-noderma lucidum polysaccharides combined with met-formin has an obvious prevention on pathological chan-ges of thoracic aorta in diabetic rats. The possible mechanism may be related to repressing oxidative stress of thoracic aorta, regulating the dyslipidemia, and the down regulation of the expression of VEGF in thoracic aorta.
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Objective To investigate the expressions of advanced glycosylation end products (AGE) in skin of mice with diabetes mellitus (DM) for different durations,and to evaluate their influence on collagen fibers.Methods Forty healthy 8-week-old male C57BL/6J mice were divided into DM group (n =20) and control group (n =20) to receive multiple intraperitoneal injections of low dose streptozotocin (50 mg/kg) and citric acid buffer (0.1 mol/L),respectively,for 5 consecutive days.Ten mice were sacrificed in each group on week 4 and 12 respectively after the last intraperitoneal injection,and full-thickness skin tissue samples were harvested from the middorsal region of each mouse.Then,hematoxylin-eosin (HE) staining was performed to observe histological changes,and total collagen content was estimated according to hydroxyproline content measured by an alkalinehydrolysis method.The cross-linking degree of collagen was determined by Edman degradation method using pepsin,the mRNA expression level of collagen type Ⅰ and Ⅲ by real-time quantitative PCR,the content of AGE by fluorospectrophotometry and Western blotting,and the level of malondialdehyde (MDA) by using a thiobarbituric acid method.Statistical analysis was carried out by t test.Results As light microscopy showed,the skin became obviously thinner in the diabetic mice with a progressive decrease in the number of collagen fibers in comparison with the control mice.On week 4 and 12 after the last injection,the diabetic mice exhibited a significant reduction in the content of hydroxyproline ((684.5 ± 76.7) vs.(787.7 ± 87.7) rg/g,(558.1 ± 73.1) vs.(757.8 ± 75.3) mg/g,both P ＜ 0.01) and in the levels of cross-linked collagen as well as mRNA expressions of collagen Ⅰ and Ⅲ (P ＜ 0.01 or 0.05),but a significant increase in the content of AGE ((37.47 ± 10.65) vs.(26.39 ± 3.74) AUF/mg hydroxyproline,(47.70 ± 5.66) vs.(29.91 ± 6.50) AUF/mg hydroxyproline,both P ＜ 0.01) and MDA ((6.62 ± 0.47) vs.(4.82 ± 0.56) μmol/L,(8.63 ± 0.36) vs.(5.15 ± 0.46) μmol/L,both P＜ 0.01) in skin tissue,compared with the control mice.The level of non-cross-linked collagen in skin tissue was also lower in the diabetic mice than in the control mice on week 12 (P ＜ 0.05).Moreover,the contents of hydroxyproline and the expression levels of collagen I in skin were significantly lower (P ＜ 0.05),but the levels of AGE and MDA were significantly higher (P ＜ 0.01) in the diabetic mice on week 12 than in those on week 4.Conclusions The characteristics of collagen fibers in skin are altered in diabetic mice when compared with normal control mice,which may be associated with increased AGE content and oxidative injury in skin.
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Objective To observe the expression and location of galactose β-1,4-glycosidic bonds (Gal β-1,4-GlcNAc) in keloid tissue,and to investigate the role of glycoprotein galactosylation in the formation of keloid.Methods This study included 10 keloid tissue specimens,7 hyperplastic scar tissue specimens,and 6 normal skin specimens.Lectin blot analysis was performed to measure the glycosylation level of glycoproteins,saturated picric acid-Sirius red staining followed by polarization microscopy to observe the type,expression and distribution of collagens in these specimens.The type Ⅰ/type Ⅲ collagen ratio was calculated.Immunofluorescence-based histochemistry was carried out by using Ricinus communis agglutinin I to analyze the expression and location of Gal β-1,4-GlcNAc in these skin samples,and double immunofluorescent staining to observe the colocalization of Gal β-1,4-GlcNAc and type Ⅰ procollagen α1.Results Compared with the normal skin tissue,the keloid tissue showed a significant increase in the expression of Gal β-1,4-GlcNAc in glycoproteins with a relative molecular mass of 30 000 to 40 000.Polarization microscopy revealed that there was a considerable expression of type Ⅰ collagen fibers,which amounted to (71.53 ± 4.03)％ in all the collagen fibers.The type Ⅰ/type Ⅲ collagen ratio was significantly higher in keloid tissue specimens than in normal tissue specimens (2.56 ± 0.53 vs.0.91 ± 0.11,P ＜0.05).Fluorescence microscopy showed that Gal β-1,4-GlcNAc was uniformly distributed in the membrane and cytoplasm of fibroblasts in keloid tissue,and the expression intensity of Gal β-1,4-GlcNAc in keloid tissue was notably stronger than that in normal skin tissue.There was a colocalization between Gal β-1,4-GlcNAc and type Ⅰ procollagen α1 in keloid tissue.Conclusions The expression of Gal β-1,4-GlcNAc,which is mainly observed in fibroblasts,is upregulated in keloid tissue,suggesting that Gal β-1,4-GlcNAc may be involved in the modulation of factors responsible for excessive fibre formation during the repair process of keloid.
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<p><b>OBJECTIVE</b>To investigate the effects of Ganoderma lucidum polysaccharides (GLPs) on advanced glycation end products (AGEs) and the receptor (RAGE) of aorta pectoralis in the T2DM rats, and explore the protective mechanism of GLPs on the aorta pectoralis.</p><p><b>METHOD</b>SD rats were fed with high-fat diet for 4 weeks and then were injected STZ (30 mg x kg(-1)) to induce the type 2 diabetic rats. Once the T2DM models were set successfully, rats were randomly divided into normal control group, diabetes model (DM) group, berberine (30 mg x kg(-1)) group, GLPs of low (GLPs-L), middle (GLPs-M) and high-dose (GLPs-H) group (GLPs were orally given 200, 400, 800 mg x kg(-1)). After 12 weeks' treatment, the content of fasting blood glucose and AGEs in serum were detected. The expressions of AGEs and RAGE in aortas pectoralis were measured both by immunohistochemistric assays and western-blot analysis.</p><p><b>RESULT</b>Compared with DM group, the content of blood glucose and AGEs in serum were significantly decreased in GLPs-H group and GLPs-M group (P < 0.01). Compared with DM group, the expressions of AGEs and RAGE in aorta pectoralis were decreased in other groups, especially in GLPs-H group (P < 0. 01).</p><p><b>CONCLUSION</b>GLPs could low blood glucose and protect aortas effectively. The mechanisms may be involved in down-regulation the expressions of AGEs and RAGE in aortal tissue.</p>
Subject(s)
Animals , Female , Male , Rats , Aorta , Metabolism , Pathology , Blood Glucose , Diabetes Mellitus, Type 2 , Metabolism , Pathology , Glycation End Products, Advanced , Metabolism , Plant Extracts , Pharmacology , Polysaccharides , Pharmacology , Random Allocation , Rats, Sprague-Dawley , Receptor for Advanced Glycation End Products , Receptors, Immunologic , Metabolism , Reishi , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Ganoderma lucidum polysaccharides (GLPs) on hemodynamic and antioxidation in the T2DM rats.</p><p><b>METHOD</b>SD rats were fed high-fat diet for 4 weeks and then were injected STZ (30 mg x kg(-1)) to induce the type 2 diabetes mellitus (T2DM). Once the T2DM models were set successfully, rats were randomized into six groups: normal group (NG), group of diabetes mellitus (DMG), groups of low dosage (GLPs-LG), middle dosage (GLPs-MG), high dosage (GLPs-HG) and berberine (BerG). They received GLPs with different dosages (200, 400, 800 mg x kg(-1)) and berberine (30 mg x kg(-1)) continually for 16 weeks. At 16th weekend, the following indices of rats were measured respectively: blood glucose, hemodynamic including LVSP, LVEDP, dp/dt(max) and -dp/dt(max) and the contents of NO, SOD, MDA, GSH-Px, CAT in cardiac tissue. Besides, myocardial ultrastructure was observed by electron microscope.</p><p><b>RESULT</b>Both the middle dosage and the high dosage of GLPs could low blood glucose effectively, and they could reduce LVEP but increase -dp/dt(max). Meanwhile, they could activate GSH-Px, CAT, SOD, NO, but reduce MDA in cardiac tissue and improve the myocardial ultrastructure. Compared to the DM group, the middle dosage, high dosage of GLPs and berberine showed significant improvement. Compared to the berberine group, the middle dosage showed the same effect, but the high dosage was more effective than berberine.</p><p><b>CONCLUSION</b>There is a confirmed action of GLPs in improving the hemodynamic and antioxidation in cardiac tissue of T2DM rats.</p>
Subject(s)
Animals , Female , Male , Rats , Antioxidants , Metabolism , Catalase , Metabolism , Diabetes Mellitus, Experimental , Drug Therapy , Metabolism , Diabetes Mellitus, Type 2 , Drug Therapy , Metabolism , Glutathione , Metabolism , Hemodynamics , Hypoglycemic Agents , Chemistry , Therapeutic Uses , Malondialdehyde , Metabolism , Microscopy, Electron, Transmission , Myocardium , Metabolism , Nitric Oxide , Metabolism , Polysaccharides , Therapeutic Uses , Rats, Sprague-Dawley , Reishi , Chemistry , Superoxide Dismutase , MetabolismABSTRACT
Objective To investigate the expressions of advanced glycation end products (AGEs) and their receptors in keloid. Methods Serum and skin tissue specimens were collected from 20 patients with keloid, 20 patients with hyperplastic scar and 20 normal human controls. Fluorospectrophotometer was used to measure the serum level of AGEs, and immunohistochemistry and Western blotting to detect the expressions of AGEs and AGER in skin tissue specimens. Results The serum level of AGEs was (0.713 ± 0.098) AU/ml and (0.699 ± 0.077) AU/ml respectively in patients with keloid and those with hypertrophic scar, significantly higher than that in normal controls (0.179 ± 0.056 AU/ml, F = 283.82, P < 0.01 ). A positive expression of AGEs and AGER was observed in tissue specimens of keloid and hyperplastic scar, but not in the control specimens. As Western blotting showed, the expressions of AGEs and AGER were higher in tissue specimens of keloid and hyperplastic scar than in the control specimens (F = 18.04, 42.80, both P < 0.05), while no significant difference between keloid and hyperplastic scar tissue specimens was observed (P> 0.05). Conclusion There is a high expression of AGEs and AGER in keloid, which may contribute to the development of keloid.
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<p><b>OBJECTIVE</b>To label embryonic stem (ES) cells with enhanced green fluorescent protein (EGFP) on the hypoxanthineguanine phosphoribosyl transferase (HPRT) gene locus for the first time to provide a convenient and efficient way for cell tracking and manipulation in the studies of transplantation and stem cell therapy.</p><p><b>METHODS</b>Homologous fragments were obtained by polymerase chain reaction (PCR), from which the gene targeting vector pHPRT-EGFP was constructed. The linearized vector was introduced into ES cells by electroporation. The G418(r)6TG(r) cell clones were obtained after selection with G418 and 6TG media. The integration patterns of these resistant cell clones were identified with Southern blotting.</p><p><b>RESULTS</b>EGFP expressing ES cells on the locus of HPRT were successfully generated. They have normal properties, such as karyotype, viability and differentiation ability. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages and in differentiated cells. Cultured in suspension, the "green" ES cells aggregated and formed embryoid bodies, retaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence.</p><p><b>CONCLUSIONS</b>This generation of "green" targeted ES cells is described in an efficient protocol for obtaining the homologous fragments by PCR. Introducing the marker gene in the genome of ES cells, we should be able to manipulate them in vitro and use them as vehicles in cell-replacement therapy as well as for other biomedical and research purposes.</p>
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Animals , Mice , Cells, Cultured , Chromosome Mapping , Embryo, Mammalian , Cell Biology , Green Fluorescent Proteins , Hypoxanthine Phosphoribosyltransferase , Genetics , Luminescent Proteins , Metabolism , Recombination, Genetic , Stem Cells , Metabolism , TransgenesABSTRACT
Aim To investigate the myocardial hypertrophy,the expression of transforming growth factor ?1(TGF-?1),Smad3 and Smad7 proteins in spontaneous hypertensive rats(SHR)and the effects of benazepril and candesartan.Methods SHRs of 12 weeks old were given benazepril and candesartan for 12 weeks.The tail arterial pressure was measured every two weeks.At 12 th weekend,cardiac configuration,heart mass index,area of cadiocytes,concertrations of AngⅡin plasma and myocardium,expressions of TGF-?1、Smad3 and Smad7 proteins were measured respectively.Results The arterial pressure,wall thickness,heart mass index,area of cardiocytes and the expressions of TGF-?1,Smad3 proteins increased in SHRs but were attenuated after the treatment of benazepril or candesartan.After the combined treatment,the synergistic effect could be observed.The levels of cardiac tissue and plasma AngⅡwere reduced.The expressions of Smad 7 were up-regulated after the treatment of benazepril or candesartan,while they were stable after the combined use.Conclusion There is a synergistic effect of attenuating myocardial hypertrophy in SHRs by combined use of benazepril and candesartan.It may be related to the regulation of Ang Ⅱ,decreasing the expressions of TGF-?1 and Smad3.
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Aim To investigate the effect of irbesartan and spironolactone on expressions of connective tissue growth factor(CTGF),P-p38MAPK proteins during vascular remodeling in renovascular hypertensive rats(RHR).Methods Renovascular hypertension was induced by two kidney one clip(2K1C)operation.8 weeks later,RHRs were given irbesartan or(and)spironolactone for 8 weeks.After 8 weekstreatment,the morphometric measurements were performed in the mesenteric arterioles.Concertration of carboxyterminal propeptide of typeⅠprocollagen(PⅠCP)and N-terminal propeptide of type Ⅲ procollagen(PⅢNP)in blood serum was measured by enzyme linked immunosorbent assay method,and the media collagen area was assessed by collagne-specific Picro-sirius red staining with computerized video processing.Expressions of collagen type I,CTGF,P-p38MAPK proteins were detected using immunohistochemistry respectively.Results The arterial systolic pressure was attenuated significantly after the treatment of irbesartan,and this effect could not be enhanced by spironolactone.The media thickness over lumen ratio,media cross-sectional area over luminal area ratio of mesenteric arterioles,concertration of PⅠCP and PⅢNP,media collagen area,and expression of collagen typeⅠwere significantly increased in RHRs but ameliorated by irbesartan and spironolactone.Meanwhile,the expressions of CTGF,P-p38MAPK proteins were up-regulated in RHRs but blunted significantly after the treatment of irbesartan and spironolactone.The combined treatment had the synergic effects.Conclusions There is a synergistic effect of attenuating extracellular matrix(ECM)producing and amelioration of vascular remodeling(VR)by combined use of irbesartan and spironolactone.It maybe related to the expressions of CTGF and P-p38MAPK proteins down regulated by these two drugs.