Increased Deposition Of Collagen Fibers Research Articles

Effects of moxibustion on visceral hypersensitivity and colonic inflammatory response in mice with chronic ulcerative colitis.

To observe the effects of moxibustion at "Zusanli" (ST36) on the expression levels of tumor necrosis factor (TNF)-α, TNF receptor 1 (TNF-R1), p38 mitogen-activated protein kinase (P38 MAPK), and transient receptor potential vanilloid 1 (TRPV1) in the colon tissue of mice with chronic ulcerative colitis (UC), so as to explore the underlying mechanisms of moxibustion in improving visceral hypersensitivity in chronic UC. Male C57BL/6J mice were randomly divided into normal group, normal with moxibustion (NM) group, model group, and model with moxibustion (MM) group, with 10 mice in each group. The chronic UC model was established by drinking 2.5% dextran sodium sulfate for 3 cycles. Mice in the NM and MM groups received moxibustion at ST36 for 20 min, 5 days per week with a 2-day break, for a total of 4 weeks. The disease activity index (DAI) score of each group was evaluated before and after treatment. The minimum volume threshold of abdominal wall retraction reflex (AWR) was measured to observe the intestinal sensitivity of mice. The colon length was measured. The pathological changes of colon tissue were observed by HE staining. The expression of mucin in colon goblet cells was detected by periodate Scheff staining. The intestinal fibrosis was observed by Masson staining. The number of trypsin-positive cells (i.e., mast cell) and the expression level of TNF-α in colon tissue were detected by immunofluorescence staining. The expression levels of TNF-R1, P38 MAPK and TRPV1 in colon tissue were detected by immunohistochemistry. Compared with the normal group after treatment, the model group showed increased DAI score (P<0.001), decreased AWR minimum volume threshold (P<0.01), shortened colon length (P<0.001), significant inflammatory infiltration in the colon tissue, reduced mucin secretion (P<0.01), increased collagen fiber deposition (P<0.001), and elevated expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (P<0.001, P<0.01, P<0.05). Compared with the model group, the MM group showed decreased DAI score (P<0.01), increased AWR minimum volume threshold (P<0.001), elongated colon length (P<0.001), reduced inflammatory cell infiltration, improved integrity of mucosal glandular structure, enhanced mucin secretion (P<0.01), decreased collagen fiber deposition (P<0.001), decreased number of mast cells in the colon tissue (P<0.001), and decreased expression levels of TNF-α, TNF-R1, P38 MAPK, and TRPV1 (P<0.001, P<0.01, P<0.05). There were no significant differences in the above index between the NM group and the normal group. Moxibustion can reduce visceral hypersensitivity, alleviate inflammatory infiltration and fibrotic damage in the colon tissue of mice with chronic UC. These effects may be associated with the down-regulation of TNF-α, TNF-R1, P38 MAPK, and TRPV1 expression in colon.

Intervention of modified Shenling Baizhu San on peritoneal fibrosis induced by peritoneal dialysate with different sugar concentration in rats

To investigate the effect of modified Shenling Baizhu San on the pathological changes and extracellular matrix (ECM) in rats with peritoneal fibrosis induced by peritoneal dialysate fluid (PDF) with different sugar concentration and its mechanism. Seventy male Sprague-Dawley (SD) rats were randomly divided into control group, different sugar content PDF model groups and corresponding traditional Chinese medicine intervention groups, with 10 rats in each group. Peritoneal fibrosis model was reproduced by intraperitoneal injection of 100 mL×kg-1×d-1 PDF containing 1.5%, 2.5% and 4.25% sugar once a day for 8 weeks. The rats in the control group were given the same amount of normal saline. The rats in the traditional Chinese medicine intervention groups were treated with gavage of 10 mL/kg of modified Shenling Baizhu San (containing 2.014 g crude drug per liter) immediately after modeling. The PDF model groups and the control group were given the same amount of normal saline by gavage. After 8 weeks, the peritoneal ultrafiltration volume of rats in each group was measured. The peritoneal tissues were collected and stained with hematoxylin-eosin (HE), and the structural changes and thickness of the parietal peritoneum were observed under a light microscope. After Masson staining, the deposition of collagen fibers was observed under a light microscope. Western blotting was used to detect the protein expressions of E-cadherin, α-smooth muscle actin (α-SMA) and Vimentin, the main components of ECM in parietal peritoneum. The positive expressions of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and transforming growth factor-β1 (TGF-β1) were detected by immunohistochemical staining. Compared with the control group, PDF with different sugar contents could induce peritoneal fibrosis in rats, and the degree of fibrosis was gradually aggravated with the increase of sugar content, which was manifested as peritoneal thickening, increased collagen fiber deposition, decreased peritoneal ultrafiltration volume, down-regulated expressions of E-cadherin and MMP-9 in peritoneal tissue, and up-regulated expressions of α-SMA, Vimentin, TIMP-1 and TGF-β1, and the pathological changes and ECM accumulation in peritoneal tissues were more serious in 4.25% PDF model group. After the intervention of modified Shenling Baizhu San, compared with the corresponding PDF model groups, the peritoneal fibrosis of rats was improved to varying degrees, and the effect of the 4.25% PDF+traditional Chinese medicine intervention group was more significant, the parietal peritoneum was significantly thinner (μm: 101.86±16.01 vs. 140.65±10.13, P < 0.05), collagen fiber deposition was significantly reduced, peritoneal ultrafiltration volume was significantly increased (mL: -0.01±3.45 vs. -3.53±1.84, P < 0.05), the expressions of E-cadherin and MMP-9 in peritoneal tissues were significantly up-regulated [E-cadherin protein (E-cadherin/β-actin): 0.84±0.08 vs. 0.28±0.05, MMP-9 (A value): 0.60±0.15 vs. 0.37±0.01, both P < 0.05], and the expressions of α-SMA, Vimentin, TIMP-1 and TGF-β1 were significantly down-regulated [α-SMA protein (α-SMA/β-actin): 0.36±0.08 vs. 1.05±0.09, Vimentin protein (Vimentin/β-actin): 0.53±0.07 vs. 1.19±0.04, TIMP-1 (A value): 0.49±0.06 vs. 0.87±0.02, TGF-β1 (A value): 0.67±0.04 vs. 0.89±0.10, all P < 0.05]. The degree of peritoneal fibrosis gradually increased with the increase of PDF sugar content in rats. Modified Shenling Baizhu San can improve peritoneal fibrosis induced by PDF with different sugar contents in rats, and the mechanism is related to the changes in the expression of fibrosis markers and ECM accumulation.

Kombucha tea improves glucose tolerance and reduces hepatic steatosis in obese mice

Nonalcoholic fatty liver disease (NAFLD), often associated with obesity, is becoming one of the most common liver diseases worldwide. It is estimated to affect one billion individuals and may be present in approximately 25% of the population globally. NAFLD is viewed as a hepatic manifestation of metabolic syndrome, with humans and animal models presenting dyslipidemia, hypertension, and diabetes. The gut-liver axis has been considered the main pathogenesis branch for NAFLD development. Considering that foods or beverages could modulate the gastrointestinal tract, immune system, energy homeostasis regulation, and even the gut-liver axis, we conducted an exploratory study to analyze the effects of kombucha probiotic on hepatic steatosis, glucose tolerance, and hepatic enzymes involved in carbohydrate and fat metabolism using a pre-clinical model. The diet-induced obese mice presented glucose intolerance, hyperinsulinemia, hepatic steatosis, increased collagen fiber deposition in liver vascular spaces, and upregulated TNF-alpha and SREBP-1 gene expression. Mice receiving the kombucha supplement displayed improved glucose tolerance, reduced hyperinsulinemia, decreased citrate synthase and phosphofructokinase-1 enzyme activities, downregulated G-protein-coupled bile acid receptor, also known as TGR5, and farnesol X receptor gene expression, and attenuated steatosis and hepatic collagen fiber deposition. The improvement in glucose tolerance was accompanied by the recovery of acute insulin-induced liver AKT serine phosphorylation. Thus, it is possible to conclude that this probiotic drink has a beneficial effect in reducing the metabolic alterations associated with diet-induced obesity. This probiotic beverage deserves an extension of studies to confirm or refute its potentially beneficial effects.

Role of H2S Supplementation on Burn Wound Healing and Molecular Chaperones

Treatment of non-healing burn injuries is a major challenge for the current scientific research. Hydrogen sulfide (H2S) is an endogenous gasotransmitter, which regulates redox homeostasis and cytoprotection during pathophysiological conditions. Similarly, heat shock proteins (HSPs) are molecular chaperones, which also confer cytoprotection during the wound repair process. Notably, the role of H2S as a regulator of HSPs during burn wound healing is still elusive. The present study investigated the effects of H2S supplementation on molecular chaperones during full-thickness, third-degree burn wound healing in the experimental rats. The animals were treated with sodium hydrosulphide (NaHS) as H2S donor (5 mg/kg body weight, intraperitoneal) daily for 10 days prior to burn-induction and continued till the fifth-day post-wounding. Histopathological analysis (Masson’s trichrome) revealed enhanced wound healing evident by increased collagen fiber deposition, cellular proliferation and re-epithelialisation in NaHS administered group as compared to the burn control. Furthermore, immunoblot analyses demonstrated significantly increased protein expression of molecular chaperons viz. HSP90, HSP70, HSP27, and GRP78 in H2S treated group as compared to control. Therefore, the present study signifies that H2S supplementation upregulates the protein expression levels of molecular chaperones, which could facilitate the cytoprotection during the tissue repair process and accelerates the burn wound healing.

Effect of Induced hypoglycemia on Postnatal Heart Structure in Male Albino Rats

Background: Hypoglycemia is a life-threatening stress, often seen in neonatal age because of inborn hyperinsulinism, inherited disorder in sugar metabolism and vigorous insulin treatment for juvenile-onset Diabetes Mellitus. Recurrent hypoglycemia typically disturbs cardiovascular function. Therefore this study aimed to explore the consequences of hypoglycemia on the postnatal heart structure in male albino rats.Material and Methods: Thirty six male albino rat pups at postnatal week 2(PNW2) were randomly allocated into two groups; control (GroupI) and treated (GroupII). GroupI was further split equally into control negative (gained no medicines) and control positive (injected Sc with 0.3 ml saline 3 times/week). Group II were injected Sc with human regular insulin 3 IU/kg body weight, 3 times/ week. Each of the previous groups were extra split according to age of animal sacrificing (at either PNW3 or PNW11) into 2 subgroups. Heart specimens were managed for light microscope, immuno-histochemical and morphometric studies. Blood samples were got for cardiac enzyme estimation.Results: Induced hypoglycemia resulted in histological changes in the normal heart structure manifested by apparently disarrayed hypertrophied cardiac muscle fibers, vascular congestion and increased connective tissue cells. Also, the cardiomyocyte cytoplasm exhibited cytoplasmic vacuolization and rarefaction. Moreover, nuclear pyknosis and pleomorphism were obvious. Comparable to control groups, increased collagen fiber deposition, declined glycogen content, weakened desmin and increased P53 immune expressing, elevated serum cardiac and decreased antioxidant enzymatic levels were manifested. These findings were evident at PNW11 hypoglycemic group.Conclusion: Intermittent hypoglycemia had detrimental consequences on the postnatal heart structure via oxidative stress.

Dugotrajna primena hlorokina i etanola izaziva ozbiljna morfološka oštećenja bubrega kod pacova na niskoproteinskom režimu ishrane

The aim of the study was to investigate the microscopic renal changes resulting from the concurrent administration of chloroquine and ethanol, with inadequate dietary protein using rats. Sixty-four rats were randomly distributed into eight groups of eight rats each: control groups on normal protein (NPC) or low protein diet (LPC); chloroquine treatment groups on normal protein (NPQ) or low protein diet (LPQ); ethanol treatment groups on normal protein (NPE) or low protein diet (LPE); concurrent chloroquine and ethanol treatment groups on normal protein (NPQE) or low protein diet (LPQE). Chloroquine in 0.9% normal saline was administered weekly to NPQ, LPQ, NPQE, and LPQE. While NPE, LPE, NPQE and LPQE received 6% ethanol in drinking water ad libitum, NPC and LPC received 0.9% normal saline and plain drinking water. After treatment, routine haematoxylin and eosin stain, Masson's trichrome stain for collagen, kidney volume estimation, glomeruli count, immunofluorescence for aquaporin 2 and urine volume estimation were conducted. The results showed a decreased kidney volume in all the experimental groups compared to the control. There was increased collagen fiber deposition and distortion of renal histology in the experimental groups compared to control. Concurrent administration of chloroquine and alcohol causes distortion of kidney histology and derangements of renal function in the low protein-fed rats and can cause kidney failure.

Decreased autophagy of vascular smooth muscle cells was involved in hyperhomocysteinemia-induced vascular ageing.

Ageing and hyperhomocysteinemia (HHcy) are important risk factors for cardiovascular diseases (CVDs). HHcy affects the occurrence of vascular diseases in the elderly. So far, the mechanism of HHcy-induced vascular ageing remains largely unknown. Autophagy level is significantly reduced in the ageing process, and restoring impaired autophagy to a normal state may be one of the possible ways to extend the expected longevity and lifespan in the future. In this study, we established the HHcy rat model by feeding a 2.5% methionine diet. Small animal ultrasound and the tail-cuff method indicated that the vascular pulse wave velocity (PWV) and pulse pressure (PP) of HHcy rats were increased significantly compared with the control group. Vascular morphology and structure have been changed in HHcy rats, including lumen dilation, increased collagen fibre deposition and increased p53/p21/p16 expression. In vitro, under the stimulation of homocysteine (500μmol/L, 24hours), the rat vascular smooth muscle cells (VSMCs) presented senescence, which was characterized by the increased expression of ageing-related markers, such as p16, p21 and p53 as well as increased senescence-associated beta-galactosidase (SA-β-gal) activity. Meanwhile, the autophagy level was decreased both in vivo and in vitro, shown as the increased level of autophagy substrate p62 and the reduced level of autophagy marker LC3 II/I in the thoracic aorta of HHcy rats and in Hcy-treated VSMCs, respectively. The senescence phenotype of VSMCs was reversed by increased autophagy levels induced by rapamycin. Our findings indicate that decreased autophagy of VSMCs is involved in hyperhomocysteinemia-induced vascular ageing.

Effect of fluoxetine on the testes of adult albino rats and the possible protective role of curcumin

Fluoxetine (FLX) is extensively used for the treatment of a diversity of psychiatric disorders, mainly depression. However, it can adversely affect male fertility. This study was done to clarify the changes which take place in the testes after the oral administration of FLX and to evaluate the possible preventative role of curcumin. Seventy-six adult male albino rats were randomly divided into four equal groups. Control group: kept without any treatment. Curcumin group: received daily dose of curcumin (150mg/kg body weight) through oral gavage for 8weeks. FLX group. They were given daily dose of FLX (10mg/kg body weight) given through oral gavage for 8weeks. FLX and curcumin group. They were given FLX together with curcumin with the same previous doses through oral gavage daily for 8weeks. By the end of the experiment, blood samples were collected for the biochemical study of testosterone. All the animals were anaesthetized by ether inhalation, and the testis specimens were dissected out and weighed. The specimens were subjected to histopathological, immunohistochemical, and morphometrical evaluation. FLX decreased serum testosterone, diminished both epithelial height and diameter of seminiferous tubules, increased collagen fiber deposition in testicular tissue and induced positive immune reaction to B-cell lymphoma-2-associated X protein. In the FLX and curcumin group, the FLX-induced changes were less remarkable. Exposure to FLX led to pronounced testicular alterations. Co-administration of curcumin with FLX ameliorated these changes.

LKB1 regulates epithelial-mesenchymal transition in Peutz-Jeghers hamartoma and intestinal epithelial cells

To investigate the molecular mechanism by which LKB1 regulates epithelial-mesenchymal transition (EMT) in Peutz-Jeghers hamartoma and intestinal epithelial cells. Immunohistochemistry was used to detect gene expression of LKB1, E-cadherin, and vimentin in 20 hamartoma tissues and 10 normal intestinal tissues, and collagen fiber deposition was analyzed using Masson trichrome staining. Normal intestinal epithelial NCM460 cells were transfected with LKB1 shRNA plasmid or negative control via lentiviral vectors, and the role of LKB1 in cell polarization and migration were determined using CCK8 and Transwell assays. Western blotting, quantitative real-time PCR (qPCR) and immunofluorescence were used to assess the alterations of EMT markers in the cells with LKB1 knockdown. Compared with normal intestinal tissues, hamartoma polyps showed significantly decreased LKB1 and E-cadherin expressions and increased vimentin expression with increased collagen fiber deposition. The cells with LKB1 knockdown exhibited enhanced cell proliferation and migration activities (P<0.01). Western blot analysis, qPCR and immunofluorescence all detected decreased E-cadherin and increased N-cadherin, vimentin, Snail, and Slug expressions in the cells with LKB1 knockdown. s LKB1 deficiency triggers EMT in intestinal epithelial cells and Peutz-Jeghers hamartoma, suggesting that EMT can serve as the therapeutic target for treatment of Peutz-Jeghers syndrome.

Histological study on the effect of rosuvastatin (Crestor) on the skeletal muscle of adult male albino rats and the possible protective effect of coenzyme Q10

Objective The aim of this study was to evaluate the effect of rosuvastatin drug on the histological structure of the skeletal muscle fiber of adult male albino rats and the possible protective role of coenzyme Q10. Background Rosuvastatin has been proven to be effective in improving serum lipid profiles. It decreases the risk for mortality in patients with coronary heart disease. It was reported that some patients treated with various statins have developed symptoms of myopathy. Coenzyme Q10 has a powerful antioxidant activity and affects membrane stability in many tissues, including skeletal muscle. Materials and methods A total of 54 adult male albino rats were used in this study and divided into four groups: group I rats served as negative controls, group II rats served as positive controls, group III rats were treated with rosuvastatin orally for 4 and 12 weeks, and group IV rats were treated with rosuvastatin and Coenzyme Q10 orally for 4 and 12 weeks. The gastrocnemius muscle was dissected and prepared for light and electron microscopic study. Results The light microscopic study of the gastrocnemius muscle of rats treated with a high therapeutic dose of rosuvastatin for 4 and 12 weeks (group III) showed variation in size, mononuclear cellular infiltration, splitting, and focal degeneration of myofibers with increased collagen fiber deposition between muscle fibers. Electron microscopic study showed mitochondrial accumulation between myofibrils and in the subsarcolemmal space, mitochondrial degeneration, and dilatation of sarcoplasmic reticulum cisterna. Coadministration of coenzyme Q10 with rosuvastatin for 4 and 12 weeks ameliorated most of the above-mentioned histological alterations in the rat skeletal muscles. Conclusion Rosuvastatin drug caused skeletal muscle fiber damage. Coenzyme Q10 leads to the protection of the skeletal muscle fibers when given concomitantly with rosuvastatin.

Periostin in Mature Stage Localized Scleroderma.

BackgroundPeriostin is a novel matricellular protein expressed in many tissues, including bone, periodontal ligament, and skin. Although its expression is prominent in various fibrotic conditions, studies of periostin in localized scleroderma are rare.ObjectiveTo investigate the expression of periostin and other molecules in localized scleroderma.MethodsA retrospective study of 14 patients with confirmed mature stage localized scleroderma was undertaken. Fourteen age-matched and biopsy site-matched subjects with normal skin were included as controls. Collagen fiber deposition, periostin, procollagen, transforming growth factor-β, and matrix metalloproteinase (MMP)-1 expression were assessed and compared between the two groups. Co-localization of α-smooth muscle actin and periostin was evaluated using confocal microscopy.ResultsPeriostin was predominantly expressed along the dermo-epidermal junction in the controls. Conversely, patients with localized scleroderma demonstrated increased collagen fiber deposition and periostin expression that was more widely distributed along the entire dermis. MMP-1 staining showed increased expression in the epidermis and dermis of patients compared to scanty expression in the controls. A semi-quantitative evaluation showed a higher proportion of excessive collagen bundle deposition (57.1% vs. 7.1%, p=0.013), diffuse periostin positivity (42.9% vs. 0%, p=0.016), and moderate MMP-1 positivity (71.4% vs. 7.1%, p=0.001) in patients than in the controls.ConclusionCompared to the controls, patients with localized scleroderma had enhanced periostin expression corresponding to increased collagen fiber deposition and unexpected overexpression of MMP-1. The results of this human in vivo study may implicate the pathogenesis of localized scleroderma.

Can vitamin E protect the pancreas against ethanol-induced alterations in adult male albino rats? A histomorphometric and immunohistochemical study

Background Ethanol has a destructive effect on the pancreas by inducing oxidative damage on pancreatic cells. This injurious effect could be ameliorated by the administration of vitamin E. Aim The aim of the study was to investigate the possible protective role of vitamin E on ethanol-induced alterations in the pancreas in adult male albino rats and also the effect of ethanol withdrawal. Materials and methods Forty adult male albino rats were divided into four equal groups (10 rats each): group I (control group), group II (ethanol-treated group), group III (ethanol and vitamin E-treated group), and group IV (recovery group). At the end of the experiment, the rats were sacrificed by cervical dislocation and their pancreas were excised and prepared for light microscopic and histochemical study. Detection of interleukin (IL)-1B and IL-6 by real-time PCR and proliferating cell nuclear antigen immunoexpression in the pancreatic tissue was carried out. Results Group II showed loss of acinar tissue, distorted degenerated pancreatic acini, dilated distorted interlobular ducts, distorted islets of Langerhans, and dilatation and congestion of the blood vessels. Interlobular inflammatory cellular infiltration was also detected. There was an increased collagen fiber deposition and diminution of the proliferating stem cells. There was a marked increase in IL-1B and IL-6 levels. Group III showed an obvious improvement in the structure of the pancreas. The recovery group showed improvement in the structure of the pancreas and minimal collagen deposition. Conclusion Vitamin E could protect the pancreas against ethanol-induced alteration in adult male albino rats.

Inflammation, oxidative stress and apoptosis cascade implications in bisphenol A-induced liver fibrosis in male rats.

Bisphenol A (BPA) is a key monomer in the production of plastics. It has been shown to be hepatotoxic. Inflammation and oxidative stress are closely linked with liver fibrosis, the major contributing factor to hepatic failure. Therefore, the aim of this study was to evaluate the impact of chronic exposure to BPA on the development of hepatic fibrosis in male rats and to determine the cross-talk between the hepatic cytokine network, oxidative stress and apoptosis. For this purpose, 30 male Wistar albino rats were divided into three equal groups as follows: the first group was given no treatment (normal control group); the second group was given corn oil once daily by oral gavage for 8weeks (vehicle control group); and the third group received BPA (50mg/kg body weight/day, p.o.) for 8weeks. BPA administration induced liver fibrosis as reflected in an increase in serum hepatic enzymes activities, hepatic hydroxyproline content and histopathological changes particularly increased collagen fibre deposition around the portal tract. In addition, there was inflammation (as reflected in increase in interleukin-1beta 'IL-1β', decrease in interleukin-10 'IL-10' serum levels and increase in IL-1β/IL-10 ratio), oxidative stress (as reflected in increase in malondialdehyde (MDA) level, reduction in reduced glutathione (GSH) content and inhibition of catalase (CAT) activity) and apoptosis [as reflected in an increase in caspase-3 level and a decrease in numbers of B-cell lymphoma 2 (BCL2)-immunopositive hepatocytes]. Interestingly, BPA had an upregulating effect on an extracellular matrix turnover gene [as reflected in matrix metalloproteinase-9 (MMP-9)] and a downregulating effect on its inhibitor gene [as reflected in tissue inhibitor of matrix metalloproteinase-2 (TIMP-2)] expression. Thus, the mechanism by which BPA induced liver fibrosis seems to be related to stimulation of the inflammatory response, along with oxidative stress, the apoptotic pathway and activation of extracellular matrix turnover.

Quantitative differentiation of normal and scarred tissues using second-harmonic generation microscopy.

The aim of this study was to differentiate normal and scarred hamster cheek pouch samples by applying a quantitative image analysis technique for determining collagen fiber direction and density in second-harmonic generation microscopy images. This paper presents a collagen tissue analysis of scarred cheek pouches of four adult male Golden Syrian hamsters as an animal model for vocal fold scarring. One cheek pouch was scarred using an electrocautery unit and the other cheek was used as a control for each hamster. A home-built upright microscope and a compact ultrafast fiber laser were used to acquire depth resolved epi-collected second-harmonic generation images of collagen fibers. To quantify the average fiber direction and fiber density in each image, we applied two-dimensional Fourier analysis and intensity thresholding at five different locations for each control and scarred tissue sample, respectively. The resultant depth-resolved average fiber direction variance for scarred hamster cheek pouches (0.61 ± 0.03) was significantly lower (p < 0.05) than control tissue (0.73 ± 0.04), indicating increased fiber alignment within the scar. Depth-resolved average voxel density measurements indicated scarred tissues contained greater (p < 0.005) fiber density (0.72 ± 0.09) compared to controls (0.18 ± 0.03). In the present study, image analysis of both fiber alignment and density from depth-resolved second-harmonic generation images in epi-detection mode enabled the quantification of the increased collagen fiber deposition and alignment typically observed in fibrosis. The epi-detection geometry is the only viable method for in vivo imaging as well as imaging thick turbid tissues. These quantitative endpoints, clearly differentiating between control and scarred hamster cheek pouches, provide an objective means to characterize the extent of vocal fold scarring in vivo in preclinical and clinical research. In particular, this non-invasive method offers advantages for monitoring scar treatments in live animals and following the effects of scarring-related treatments such as application of steroids or drugs targeting pathways involved in fibrosis. SCANNING 38:684-693, 2016. © 2016 Wiley Periodicals, Inc.

Interferon-β inhibits glioma angiogenesis through downregulation of vascular endothelial growth factor and upregulation of interferon inducible protein 10.

Interferon-β (IFN-β) has been used clinically for malignant glioma growth inhibition. Recently IFN-β is re-evaluated for its sensitization mechanism to the chemotherapeutic agent temozolomide, because angiogenesis is essential for malignant glioma growth. In this study, we investigated new mechanisms of inhibition of glioma angiogenesis by IFN-β. Three malignant glioma cell lines, U87, TK2 and Becker, were used for in vitro study. The effect of IFN-β for these cell lines were evaluated by means of proliferation (MTT assay), conditioned medium induced HUVEC migration, VEGF and interferon inducible protein 10 (IP10, angiogenesis inhibitor) expression by RT-PCR and western blot analysis. SCID mouse U87 subcutaneous model and U87 implant cranial window model were used for in vivo study. The effect of IFN-β with the models was evaluated by means of tumor growth, tumor tissue expression for VEGF and IP10, tumor tissue CD31 positive vessel densities, apoptosis and tumor microcirculation (blood velocity, interaction between leukocytes and endothelial cells). In vitro, IFN-β upregulated IP10 expression and downregulated VEGF expression time- (4–48 h) and dose- (10–5,000 U/ml) dependently. At the same dose, glioma cell-induced HUVEC migration was inhibited, but cell proliferation was not affected. IFN-β local and systemic injection at 105 U and at 5×105 U/day, for 15 days inhibited U87 subcutaneous growth significantly. In the tumor tissues, VEGF expression and vessel densities were downregulated, but IP10 expression and apoptosis index upregulated. In addition, IFN-β local injection increased collagen fiber deposition in the tumor tissues. IFN-β 5×105 U/day, s.c. injection for 7 days reversed the decreased leukocyte adhesion to endothelial cells, but did not affect blood velocity and vessel images. One of the important roles of IFN-β for malignant glioma growth inhibition was anti-angiogenesis by directly inhibiting angiogenesis through downregulation of VEGF and upregulation of IP-10 and indirectly changing the tumor microcirculation and regulating the interstitial pressure.

Effect of green tea on aged rat skeletal muscle

Background Age-related sarcopenia represents a powerful risk factor for the loss of independence and physical disability in the elderly. Green tea contains a series of polyphenols called catechins and has been applied for disease prevention and treatment. Aim of the study To determine the effects of the intake of green tea on age-related changes in skeletal muscle. Materials and methods Thirty female albino rats were divided into three equal groups: group I (control adults) included 6-month-old rats, group II (aged) included 18-month-old rats, and group III (green tea treated) included 12-month-old rats that received green tea at a dose of 300mg/kg body weight in 1.5ml distilled water orally daily by a gastric tube for 6 months. Quadriceps muscle was dissected bilaterally and prepared for light and electron microscopical examination. Morphometric and statistical studies of the mean cross-sectional area of myofibers were carried out in the three studied groups. Results With increasing age, there were focal degeneration and atrophy, and significant decrease in the mean cross-sectional area of myofibers. The connective tissue framework showed increased collagen fiber deposition and deficient reticular fibers. Ultrathin sections showed areas of myofibrillar loss and mitochondria with destroyed cristae. Dense shrunken nuclei of satellite cells were observed. After treatment with green tea, normal-appearing myofibers, except for centrally located nuclei, were observed. The mean cross-sectional area of myofibers increased significantly compared with the aged group. Collagen and reticular fibers were relatively similar to those of the control. Proliferation of satellite cells was observed with restoration of normal myofiber ultrastructure. Conclusion Good regenerative outcome was observed in aged skeletal muscle after the intake of green tea.

Time-course of iron overload and biochemical, histopathological and ultrastructural evidence of pancreatic damage in hypotransferrinaemic mice.

1. The time course of iron overload of the pancreas in hypotransferrinaemic mice maintained on a standard rodent diet was compared with biochemical and histological markers of tissue damage. 2. Pancreatic iron levels increased linearly from weaning till 9 months of age [73.3 nmol/mg of tissue (SEM 9.9; n = 5) compared with 0.9 nmol/mg of tissue (SEM 0.1; n = 4) in age-matched controls] then decreased linearly till at least 18 months of age. 3. Investigation of tissue distribution of newly absorbed radioiron suggested that significant redistribution of iron from liver to pancreas (rather than direct dietary iron sources) must be invoked to explain the rate of pancreatic iron loading in hypotransferrinaemic mice. 4. Pancreatic epithelial cells first showed altered morphology at 9 months of age. At 12 months of age, the pancreatic epithelium had developed a micronodular appearance, with large numbers of acini replaced by atrophic, degenerated acinar cells. Increased collagen fibre deposition was evident by trichrome staining and by electron microscopy. Biochemical markers of pancreatitis (serum lipase, tissue pancreatitis-associated protein mRNA) were elevated before 9 months of age, whereas the levels of pancreatic amylase mRNA declined from 9 months of age. 5. The data suggest that iron loading of hypotransferrinaemic mouse pancreas proceeds up to a threshold level at 9 months of age followed by a progressive atrophy of secretory epithelium. The hypotransferrinaemic mouse pancreas is a useful model system for investigation of parenchymal cell damage by iron.