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Objective: To investigate the effects of C-21 steroidal glucosides from Cynanchum auriculatum on renal fibrosis in rats caused by unilateral ureteral ligation and explore its mechanisms. Methods: A total of 15 rats were randomly chosen as sham operation group (Sham), while the remaining rats underwent unilateral ureteral ligation. The rats after the operation were randomly divided into four groups, namely the Sham operation group, the model group, the positive control group (20 mg/kg), C. auriculatum high group (400 mg/kg), C. auriculatum low group (200 mg/kg), 15 per group, and the rats in each group were administrated with corresponding drugs. After the intervention of 28 d, all the rats were sacrificed and the kidneys were then removed. The content of hydroxyproline was measured. HE and Masson staining were conducted to assess kidney pathological changes and renal fibrosis. The protein expression of TGF-β1, α-SMA, and E-cadherin were tested with immunohistochemistry and Western blotting. The mRNA expression of collagen-I and collagen-III was evaluated using qRT-PCR. Results: Compared with the model group, C-21 steroid glycosides significantly alleviated the kidney pathological injury and renal fibrosis, and reduced fibrous tissue and collagen proliferation. C-21 steroid glucosides from C. auriculatum can significantly reduce the kidney/body weight ratio and the content of hydroxyproline in kidney tissue (P < 0.05, 0.01), and it showed in a dose-dependent manner. Compared with the model group, the expression of α-SMA and TGF-β1 was decreased significantly, and the expression of E-cadherin was increased significantly (P < 0.05, 0.01) in C. auriculatum group. Moreover, compared with the model group, C-21 steroidal glucosides significantly down-regulated the mRNA expression of collagen-I and collagen-III mRNA (P < 0.01). Conclusion: C-21 steroid glycosides from C. auriculatum can effectively attenuate renal fibrosis in rats with unilateral ureteral ligation and its underlying mechanisms may be related to inhibiting the over-expression of collagen-I and collagen-III, down-regulating the expression of α-SMA and TGF-β1, and up-regulating the expression of E-cadherin. By regulating the tubular epithelial-mesenchymal transition, it exerts its anti-fibrotic effect.
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ObjectiveTo investigate the expression of transforming growth factor β1 (TGF-β1) and collagen I, III (Col I, III) in vulvar lichen sclerosis (VLS) and their role in VLS.MethodsThe specimens of30 VLS tissues (15 in early stage and 15 in progressive stage), and 15 vulvar normal skin tissues were selected by biopsy or surgical excision from 2016 to 2018 in our hospital. The expression of Vimentin was detected by immunohistochemistry. The expression of TGF-β1, TGF-β2, Col I and Col III mRNA was detected through RT-PCR. Simultaneously, the fibroblasts were visualized in diseased tissues by labeling Vimentin.ResultsThe expression of Vimentin in VLS was increased significantly (P0.05). Col I mRNA was up-regulated in VLS, obviously in early stage. Meanwhile, Col III mRNA down-regulated gradually from the early to the progressive stage of VLS. Therefore, the Col I/III ratio increased gradually.ConclusionThe increase of fibroblasts and TGF-β1 in dermis of VLS promotes the synthesis of Col I and reduces the content of Col III, which may be one of the factors leading to the decrease of skin elasticity in VLS.
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Objective To observe the effect of edaravone (EDA) on the expressions of malondialdehyde (MDA), phosphorylated extracellular signal-regulated kinase (p-ERK), nuclear factor kappa-B cells (NF-κB) and Collagen-III in rat model of unilateral ureteral obstruction (UUO), and discuss the protective effect of EDA on renal-interstitial fibrosis. Methods Fifty-four male SD rats were randomly divided into three groups (18 each): sham group (normal control), UUO group and EDA group. Rats in UUO group and EDA group received UUO operation. One day before UUO operation, rats in EDA group received intraperitoneal injection of 3.75mg/(kg·d) EDA, and those in sham group and UUO group were injected with equivalent normal saline. Rats were sacrificed in three batches at the 3rd, 7th and 14th day after operation, six from each group in each batch. The renal pathological changes were observed via HE staining and Masson staining. The contents of MDA in renal tissues were determined by spectrophotometry, and the expression levels of p-ERK1/2, NF-κB and collagen-III were detected by immunohistochemistry. Results HE and Masson staining showed no significant changes of MDA content, renal tubules and interstitium in sham group at 3rd, 7th and 14th day, but obvious changes of interstitium and increased fibre deposition in UUO group and EDA group compared with sham group. Compared with UUO group, the lesions of renal tubular and interstitium significantly reduced and the interstitial fibrosis deposition decreased in EDA group. Spectrophotometry indicated that the MDA contents showed no significant difference between EDA group and UUO group [(9.261±0.496)nmol/mg prot and (10.143±0.301)nmol/mg prot] on the 3rd day after operation, but was higher than in sham group [(6.918±1.120)nmol/mg prot]. The MDA contents at the 7th and 14th day after surgery in EDA group [(11.545±0.620)nmol/mg prot and (15.203±0.512)nmol/mg prot] were significantly lower than in UUO group [(13.405±0.612)nmol/mg prot and (18.133±1.684)nmol/mg prot]. Immunohistochemistry analysis showed that no significant difference existed in the expressions of p-ERK1/2, NF-κB and Collagen III at the 3 time points in sham group; At the 3rd day after surgery, the expressions of p-ERK1/2, NF-κB and Collagen III were higher in EDA group (19 055.7±1866.1, 11 217.0±989.9 and 9724.1±341.9) than in sham group (14 100.3±1822.7, 7975.1±2058.5 and 6890.0±2389.9, P0.05). At the 7th and 14th day after surgery, the expression level of p-ERK1/2 was significantly lower in EDA group (26 228.0±523.0 and 30 647.1±583.8) than in UUO group (28 254.9±886.6 and 33 240.3±1330.4); the expression level of NF-κB was obviously lower in EDA group (16 374.4±1045.3 and 21 111.2±1022.5) than in UUO group (18 799.4±357.0 and 27 125.2±2873.3); the expression level of Collagen-III was markedly lower in EDA group (12 470.9±506.1 and 19 615.8±1120.1) than in UUO group (15 049.9±1372.3 and 22 868.9±1889.2), all the differences were of statistical significance (P<0.05). Conclusion Oxidative stress and the signaling pathway of p-ERK1/2 and NF-κB may play a role in the progress of renal interstitial fibrosis (RIF) in UUO rat model, and EDA may have an antifibrosis effect by down-regulating the levels of oxidative stress in kidney tissue and inhibiting the signaling pathway of p-ERK1/2 and NF-κB.
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AIM: To investigate the effect of streptozotocin-induced diabetes on the expression of NADPH oxidase, and to determine the effect of exercise on the expression of NADPH oxidase.METHODS: One weeks after successful modeling, the effect of diabetes mellitus on the expression of NADPH oxidase subunits in the cardiac tissue was determined.The effect of exercise for 8 weeks on the protein expression of NADPH oxidase subunits and the effect of diabetes on the protein expression of collagen in the heart were observed, and whether 8 weeks of exercise affected the expression of collagen were also measured.RESULTS: Diabetes mellitus resulted in the increased expression of NADPH oxidase subunits p47phox and gp91phox in the cardiac ventricle, and exercise for 8 weeks inhibited the increase in the expression of NADPH oxidase subunits p47phox and gp91phox.Diabetes mellitus significantly increased the expression of p47phox in the atrial muscle, while exercise inhibited this increase.Diabetes mellitus significantly increased the levels of cardiac collagen III, while exercise reduced the increase in collagen protein.CONCLUSION: Reduction of diabetic rat heart p47phox and gp91phox expression by effective exercise therapy may be an important mechanism of improving the cardiac matrix by inhibiting myocardial oxidative stress and decreasing collagen expression, thus preventing diabetic cardiomyopathy.
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Objective@#To investigate the effect of cannabinoid receptor-2 (CB2) agonist AM1241 on the mRNA and protein expression of platelet-derived growth factor (PDGF) and collagen-III (Col-III) in the liver tissue of mice with experimental liver fibrosis induced by carbon tetrachloride (CCl4).@*Methods@#Totally 38 8-week-old male C57BL/6J mice were randomly divided into control group, model group, 3 mg/kg CB2 receptor agonist (AM1241) group, and 9 mg/kg AM1241 group. All mice, except for the control group, were treated with 30% CCl4 (three times a week, 5 ml/kg body weight, 16 weeks) to establish a liver fibrosis model. Meanwhile, 3 and 9 mg/kg AM1421 was intraperitoneally injected for daily intervention, respectively. The dosage was adjusted according to actual body weight. The same solvent was given in the control group. The serum level of aspartate aminotransferase (AST) was measured by serum enzyme digestion. The liver inflammation and fibrosis were observed by HE staining of tissue slices. The mRNA and protein expression of PDGF and Col-III in hepatic tissue was determined by real-time PCR and immunohistochemistry.@*Results@#Compared with the control group, the mice in model group showed severe liver fibrosis, significantly elevated serum AST level (742 ± 300.8 U/L vs 118.1 ± 31.1 U/L, P < 0.05), and significantly increased mRNA and protein expression of PDGF and Col-III in liver tissue (P < 0.05). Compared with the model group, the mice in 3 mg/kg AM1241 group and 9 mg/kg AM1241 group had less severe liver fibrosis, and significantly reduced serum AST levels (116.6 ± 13.68 U/L vs 742 ± 300.8 U/L, P < 0.05; 113.8 ± 16.01 U/L vs 742 ± 300.8 U/L, P < 0.05) and mRNA and protein expression of PDGF and Col-III in liver tissue (P < 0.05).@*Conclusion@#CB2 receptor agonist AM1241 can inhibit the mRNA and protein expression of PDGF in the liver tissue of mice with hepatic fibrosis, and reduce extracellular matrix synthesis.
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Introduction and Objective: Because L-PRP constitutes an important source of growth factor that is associated with osteogenesis and fibrogenesis, the aim of this study was to evaluate the effect of L-PRP on the presence of collagen III and MMP-2 and MMP-9, while comparing these results by means of a histomorphometric analysis of bone matrix and fibrous deposition on bone repair. Material and methods: Four bone defects of 8 × 2 mm were created on the calvaria of 21 rabbits. The surgical defects were treated with either particulate autograft, particulate autograft mixed with L-PRP, or L-PRP alone. Animals were euthanized at 2, 4, and 6 weeks postoperative. Histomorphometric and immunohistochemical analyses were performed to assess repair time, as well as the expression of collagen III and MMPs. Results: In contrast to the results of the L-PRP-free groups, the histomorphometric results of the L-PRP groups demonstrated intense fibrotic deposition along with hindered bone matrix deposition. These results coincided with the larger occurrence of diffuse collagen III deposition and the scarce presence of MMP- 2 and -9 spread among the fibrous tissue. Conclusion: Thus, the results suggest that L-PRP not only induces an intense fibrosis rich in collagen III, which is not degraded, but also suppresses MMP-2 and -9 expressions, mimicking a similar pathological event as that of a cleft-palate or cranial suture.
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OBJECTIVE: This study investigated the effect of Low Intensity-pulsed Ultrasound (LIPUS) on the repair process of ruptured Achilles tendon using a rat model and also examined the regulation of a biological molecule that may contribute to this in vitro and in vitro. METHODS: To investigate the effect of LIPUS and its biological mechanism ofpromoting Achilles tendon repair after acute injury, ninety-eight male Sprague-Dawley (SD) rats (mean body weight, 258 ±9.8 g) aged 12 weeks were used in this study. To create the model, the Achilles tendon attachment site and musculotendinous junction were ruptured under direct vision. The leg on one side was exposed to LIPUS (frequency at 1.5 MHz, the repetition cycle at 1.0 kHz, the burst width at 200 msec and the power output at 45 mW/cm2), for 20 minutes daily with a 0.7 mm diameter probe. Results:Low Intensity-pulsed Ultrasound treatment accelerated the repair of the Achilles tendon compared to the untreated group, judged by electron microscopy. Both cyclo-oxygenase (COX)-2* and EP4* expressions were over-expressed in the LIPUS treated group in the inflammatory period, and TGFJ31* expression was markedly induced in LIPUS treated groups followed by collagen I* and III* expression in the repair and reconstitution process. CONCLUSION: These findings suggest that LIPUS is potentially able to accelerate the repair of acute ruptured Achilles tendon in several ways: by exaggerating inflammation by inducing COX-2 and EP4 and reconstituting tissue by inducing TGFJ31 followed by collagen I and III. (*: p < 0.05, **: 0.001).
OBJETIVO: Este estudio estuvo encaminado a investigar el efecto de los ultrasonidos pulsados de baja intensidad (LIPUS) sobre el proceso de reparación del tendón de Aquiles tras una ruptura, usando un modelo de rata. Asimismo, se examinó la regulación de una molécula biológica que puede contribuir a este proceso in vitro e in vitro. MÉTODOS: Con el fin de investigar el efecto de LIPUS y el mecanismo biológico por el cual este efecto promueve la reparación del tendón de Aquiles tras una lesión aguda, noventa y ocho ratas machos Sprague-Dawley (SD) (peso corporal promedio, 258 ± 9.8 g) de 12 semanas de edad fueron usadas en este estudio. Para crear el modelo, el sitio de ligazón microbiológica del tendón de Aquiles y la unión músculo-tendinosa fueron desgarrados bajo visión directa. La pierna de un lado fue expuesta a LIPUS (frecuencia de 1.5 MHz, ciclo de repetición de 1.0 kHz, ancho de ruptura de 200 msec, y potencia de salida de 45 mW/cm2), por 20 minutos diariamente con una sonda de 0.7 mm diámetro. RESULTADOS: El tratamiento de ultrasonidos pulsados de baja intensidad aceleró la reparación del tendón de Aquiles, en comparación con el grupo no tratado, según se apreció mediante el microscopio electrónico. Tanto la ciclo-oxygenasa (COX)-2* como las expresiones EP4* estuvieron sobe-expresadas en el grupo tratado con LIPUS en el periodo inflamatorio, y la expresión TGFfi1* fue marcadamente inducida en los grupos tratados con LIPUS seguidos por la expresión de colágeno I* y III* en el proceso de reparación y reconstitución. CONCLUSIÓN: Estos resultados sugieren que LIPUS puede potencialmente acelerar la reparación del tendón de Aquiles luego de un desgarramiento, de varias maneras: exagerando la inflamación mediante inducción de COX-2 y EP4 y reconstituyendo el tejido induciendo TGFfil seguido por colágeno I y III. (*: p < 0.05, **: 0.001).
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Animals , Male , Rats , Achilles Tendon/injuries , Ultrasonic Therapy/methods , Wound Healing/physiology , /metabolism , Rats, Sprague-Dawley , Rupture , Wounds and Injuries/therapyABSTRACT
OBJECTIVE: We present the difference of histopathologic changes of the internal elastic lamina (IEL) and collagen III in the superficial temporal artery (STA) between aneurysmal patients and non-aneurysmal patients. Also, the pathologic data with clinical features by comparative methods to validate the risk factor of the intracranial aneurysm are presented. METHODS: Samples of the STA were harvested form 38 patients including aneurysmal and non-aneurysmal patients undergoing craniotomy. Paraffin-embedded sections were examined, using hematoxylin and eosin, van Giebson and mouse anti-collagen III staining techniques. Histopathological observations were analysed and correlated with clinical features such as presence of aneurysm, hypertension, age, and sex. RESULTS: Twenty-seven patients had the intracranial aneurysm. Of these 24 patients were 50 years old or older. Nineteen patients had a history of hypertension. Twenty patients were female. Histopathological study demostrated the derangement of IEL and the deficiency of type III collagen were prominent in aneurysmal patients (p<0.05). Fifty years old or older patients did not show correlation with the deficiency of type III collagen, but with the derangement of IEL (p<0.05). The female sex was not correlated with the derangement of IEL but with the deficiency of type III collagen (p<0.05). However, Hypertension was not correlated with these pathologic data. CONCLUSION: Patients with intracranial aneurysms have severe histopathologic changes of the arterial wall showing the derangement of IEL and the deficiency of type III collagen. In the clinico-pathologic study, the advanced age and female sex were considered as risk factors of the intracranial aneurysm.