Immunohistochemical Expression Of TGF-β1 Research Articles

Therapeutic Effect of Melatonin on CCl4-Induced Fibrotic Liver Model by Modulating Oxidative Stress, Inflammation, and TGF-β1 Signaling Pathway in Pinealectomized Rats.

The study aimed to determine the CCl4-induced liver fibrosis model in pinealectomized rats and biochemically, immunohistochemically, and histopathologically investigate the therapeutic effect of melatonin on liver fibrosis. The surgical procedure for pinealectomy was performed at the beginning of the study, and the sham and pinealectomized rats were administered CCl4 dissolved in corn oil (1:1) alone every other day to induce liver fibrosis or together with melatonin (10 mg/kg) therapy for 15 days. Melatonin is an essential therapeutic agent and offers an alternative therapeutic strategy in CCl4-induced liver fibrosis by suppressing inflammation, oxidative stress, and the TGF-β1 signaling pathway. Treatment with melatonin ameliorated CCl4-induced liver fibrosis by restoring hepatocellular damage and reducing plasma AST, ALT, and ALP values. Melatonin increases the activity of SOD and CAT, which are important enzymes for antioxidant defence, and raises GSH levels, which further enhances antioxidant function. Also, melatonin reduced hepatic inflammation (IL-6 and IL-1β) and oxidative stress indices. Moreover, histopathological changes and immunohistochemical expression of TGF-β1 were restored following melatonin supplementation in the CCl4-induced liver fibrosis model in pinealectomized rats. Our study shows that melatonin supplementation has a beneficial effect in protecting the liver fibrosis induced by CCl4 in pinealectomized rats.

MO235: TGF-Β1/PSMAD3 Dependent Signaling in Glomerulonephritis and Its Association With Renal Injury and Progression

Abstract BACKGROUND AND AIMS Transforming growth factor-β1 (TGF-β1) has long been considered as a potent, multifunctional cytokine that is involved in the pathogenesis of fibrosis and inflammation, which acts through Smad signaling in renal pathology. We intended to investigate the expression of TGF-β1/Smad3 signaling in glomerulonephritis (GN) and to assess its role as risk factor for progression to chronic kidney disease (CKD). METHOD We evaluated the immunohistochemical expression of TGF-β1, phosphorylated Smad3 (pSmad3) and Smad7 semiquantitatively and quantitatively using computerized image analysis program in different compartments of 50 renal biopsies with GN and the results were statistically analyzed with clinicopathological parameters. We also examined the associations among their expressions, the impact of their co-expression and their role in progression to CKD. RESULTS TGF-β1 expression correlated positively with segmental glomerulosclerosis (P = .025) and creatinine level at diagnosis (p = .002), while pSmad3 expression with interstitial inflammation (P = .024). In glomerulus, we recorded different expression patterns of pSmad3 in crescents, while concomitant expressions of strong Smad7 and moderate pSmad3 were observed to be correlated with renal inflammation, such as cellular crescent (P = .011), intense interstitial inflammation (P = .029) and lower serum complement 3 (P = .028) and complement 4 (P = .029). We also reported a significant preferable expression between pSmad3 and glomerular endothelial cells of proliferative GN (P = .045) and podocytes of non-proliferative GN (P = .005). Finally, on multivariate Cox-regression analysis, TGF-β1 expression (HR = 5.08; 95% CI 1.133–22.78; P = .034) was emerged as independent predictor for CKD. CONCLUSION TGF-β1/Smad3 dependent signaling is upregulated, especially in proliferative GN, with specific characteristics in different forms of GN. TGF-β1 expression is indicated as independent risk factor for progression to CKD, while specific co-expression pattern of pSmad3 and Smad7 in glomerulus is correlated with renal inflammation.

Immunohistochemical Expression of TGF-β1 in Kidneys of Cats with Chronic Kidney Disease.

Transforming growth factor-beta 1 (TGF-β1) plays a central role in the progression of chronic kidney disease (CKD). However, in feline CKD, renal expression of TGF-β1 and how it changes as the disease progresses have not been fully studied. In the present study, we immunohistochemically assessed the renal expression levels of TGF-β1 in cats with CKD and statistically analyzed its correlation with CKD severity. Clear immunosignals were detected in the glomerular mesangial cells, Bowman’s capsules, proximal tubules, distal nephrons, platelets, and vascular smooth muscles in the kidneys of cats with CKD. Statistically, luminal signals in the distal nephrons showed positive correlations with plasma creatinine levels and glomerulosclerosis, while those in the proximal tubules and platelets showed negative correlations with plasma urea and/or creatinine levels. Therefore, it was suggested that the changes in the renal expression of TGF-β1 could be associated with progression of feline CKD.

Platelet-concentrated and platelet poor-plasma promote different pattern on immunohistochemical expression of TGF-β1, however they impairs the osteoneogensis in calvarial defects treated with autograft due suppression of osteocalcin.

ABSTRACTPurposeHerein we evaluated the effects of platelet concentrate (PC) and platelet-poor plasma (PPP) on bone repair using noncritical defects in the calvaria of rabbits and compared them to the presence of TGF-β1 and osteocalcin on reparative sites.MethodsFive noncritical defects of 8.7 mm in diameter were created on the calvaria of 15 animals. Each defect was treated differently, using autograft (ABG), ABG associated with PC (ABG + PC), ABG with PPP (ABG + PPP), isolated PPP, and blood clot (control). The animals were submitted to euthanasia on the second, fourth and sixth week post-surgery.ResultsThe defects that received ABG+PC or PPP demonstrated lower bone formation when compared to specimens that received ABG in the same period. These results coincided to significant higher immunopositivity for TGF-β1 for specimens that received PC, and lower presence of cytokine in the group PPP. However, either higher or lower presence of TGF-β1 were also correlated to lower presence of osteocalcin. Likewise, these results were similar to findings in specimens treated only with PPP when compared to control.ConclusionsPC and PPP were not effective when applied in association with ABG. Similarly, isolated use of PPP was not beneficial in optimizing the bone repair.

Immunohistochemical Expression of TGF-β1 and Ki-67 in a Paraffin-Embedded Section of Breast Cancer Tissue

We studied laboratory and medical archives during the year 2017 for 43 female patients with ages ranged between 25 and 70 years who had been primary malignant breast cancer (BC) and operated for that. We study the TGF-β 1 and Ki-67 Immunohistochemical (IHC) expression in paraffin-embedded section of BC tissue of those patients. Then we classified the patients result according to the patients ages (premenopausal, postmenopausal), size of the BC tumor (T1-T4 lymph node metastases status ((No-N3) and distant metastases (M0–1) We detected there was high expression rate of TGF-β1 & Ki67 in the BC tissue samples from patients with age < 50years (pre-menopausal age), when it compared to patients whose age ≥ 50years (postmenopausal) Also statistically significant positive association noticed between TGF-β1 & Ki67 IHC expression with the size of BC (T1-T4), lymph node(s) metastases (N0-N3) and distance metastases (M0-M1).

Prolonged use of alendronate alters the biology of cranial repair in estrogen-deficient rats' associated simultaneous immunohistochemical expression of TGF-β1+, α-ER+, and BMPR1B.

TGF-β1 is a cytokine that may induce both osteoneogenesis through Runx-2 or fibrosis via the transcription of α-smooth muscle actin (α-SMA). Because it has been previously known that alendronate increases the level of TGF-β1 and that under the usual condition of bone metabolism the estrogen may prevent the fibrotic effect of TGF-β1, the aim of this study was to evaluate if alendronate alters the cellular differentiation process post calvarial surgery in estrogen-deficient specimens. A transosseous defect that was 5mm in diameter was created on the calvarium of each of 32 female rats with previous ovarian-salpingo-oophorectomy. All defects were treated with autografts, and 16 rats received the administration of 1mg/kg of alendronate three times a week until euthanasia on the 15th and 60th day post surgery. Histomorphometric and immunohistochemical analyses of the expression of TGF-β1, estrogen receptor alpha nuclear (α-ER), α-SMA, BMPR1B, and Runx-2 were performed, and ELISA was used to measure the level of estrogen. All animals demonstrated low levels of estrogen post ovarian-salpingo-oophorectomy. The histological results demonstrated larger bone matrix deposition in specimens treated with alendronate on the 15th day post surgery. The result was associated with a higher co-expression of TGF-β1, BMPR1B, and Runx-2 when compared with the control group. In addition, on the 60th day post surgery, the increase of bone matrix deposition from 15th to 60th day was discrete in specimens treated with alendronate compared with the control group. This result coincided with the intense simultaneous expression of TGF-β1, α-ER, and α-SMA, whereas the expression of BMPR1B and Runx-2 decreased. The prolonged administration of alendronate altered the cranial repair in ovarian-salpingo-oophorectomized specimens due to the simultaneous occurrence of low estrogen and the presence of TGF-β1+/α-ER+ inducing the presence of α-SMA+, whereas BMPR1B and Runx-2 were suppressed. The prolonged administration of alendronate alters osteoneogenesis and induces an unusual microenvironment in the bone that seems to imitate the physiological tissue damage that culminates in the loss of the functional layer of endometrium.

Immunohistochemical expression of TGF-β1 and MMP-9 in periapical lesions.

The objective of this study was to evaluate the expression of matrix metalloproteinase 9 (MMP-9) and transforming growth factor beta (TGF-β1) in periapical lesion samples correlated with the intensity of the inflammatory infiltrate and thickness of the epithelial lining. Forty-five cases of periapical lesions (23 periapical granulomas and 22 radicular cysts) were subjected to morphological and immunohistochemical analyses using anti-MMP-9 and anti-TGF-β1 antibodies. The data were analyzed using the following tests: non-parametric Mann-Whitney, chi-square, Fisher's exact test and Spearman's correlation test (P<0.05). Analysis of inflammatory infiltrate revealed that 78% of periapical granulomas presented infiltrate grade III, in contrast with 32% of radicular cysts (P<0.001). Morphological evaluation of the epithelial thickness in radicular cysts revealed the presence of atrophic epithelium in 86% of the cysts. The immunostaining of MMP-9 was score 2 in 67% of the granulomas and 77% of the cysts. Both lesions were predominantly score 1 for TGF-β1. Significant differences were confirmed between the expression scores of TGF-β1 and MMP-9 in periapical granulomas (p = 0.004) and in radicular cysts (p < 0.001). Expression of TGF-β1 was different for periapical granulomas and radicular cysts. This immunoregulatory cytokine seems more representative in asymptomatic lesions. The extracellular matrix remodeling process dependent on MMP-9 seems to be similar for both periapical granulomas and radicular cysts. TGF-β1 and MMP-9 may play an important role in the maintenance of periapical lesions.

Immunohistochemical Expression of TGF-β1 and Osteonectin in engineered and Ca(OH)2-repaired human pulp tissues.

The aim of the present study was to evaluate the expression of transforming growth factor-β1 (TGF-β1) and osteonectin (ON) in pulp-like tissues developed by tissue engineering and to compare it with the expression of these proteins in pulps treated with Ca(OH)2 therapy. Tooth slices were obtained from non-carious human third molars under sterile procedures. The residual periodontal and pulp soft tissues were removed. Empty pulp spaces of the tooth slice were filled with sodium chloride particles (250-425 µm). PLLA solubilized in 5% chloroform was applied over the salt particles. The tooth slice/scaffold (TS/S) set was stored overnight and then rinsed thoroughly to wash out the salt. Scaffolds were previously sterilized with ethanol (100-70°) and washed with phosphate-buffered saline (PBS). TS/S was treated with 10% EDTA and seeded with dental pulp stem cells (DPSC). Then, TS/S was implanted into the dorsum of immunodeficient mice for 28 days. Human third molars previously treated with Ca(OH)2 for 90 days were also evaluated. Samples were prepared and submitted to histological and immunohistochemical (with anti-TGF-β1, 1:100 and anti-ON, 1:350) analyses. After 28 days, TS/S showed morphological characteristics similar to those observed in dental pulp treated with Ca(OH)2. Ca(OH)2-treated pulps showed the usual repaired pulp characteristics. In TS/S, newly formed tissues and pre-dentin was colored, which elucidated the expression of TGF-β1 and ON. Immunohistochemistry staining of Ca(OH)2-treated pulps showed the same expression patterns. The extracellular matrix displayed a fibrillar pattern under both conditions. Regenerative events in the pulp seem to follow a similar pattern of TGF-β1 and ON expression as the repair processes.

Transforming growth factor-β1 and α-smooth muscle actin in stromal fibroblasts are associated with a poor prognosis in patients with clinical stage I–IIIA nonsmall cell lung cancer after curative resection

The aims of this study were to investigate the expression of transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) in surgical resection specimens from nonsmall cell lung cancer (NSCLC) and to evaluate the prognostic significance of this gene expression in stromal fibroblasts for patients with clinical stage I-IIIA NSCLC. The immunohistochemical expression of TGF-β1 and α-SMA was evaluated in 78 formalin-fixed paraffin-embedded tumor specimens from clinical stage I-IIIA NSCLC. Correlations between this gene expression and the clinicopathologic characteristics were determined by chi-square test. The prognostic impact of this gene expression in stromal fibroblasts with regard to overall survival (OS) was determined by Kaplan-Meier and Cox hazard proportional model. The percentages of high TGF-β1 expression in stromal fibroblasts and cancer cells were 19.2 % (15/78) and 35.9 % (28/78), respectively. There were 28.2 % (22/78) of patients with high α-SMA expression in stromal fibroblasts. The analysis revealed a significant positive association between TGF-β1 expression in stromal fibroblasts and in cancer cells (χ (2) = 4.86, p = 0.03). No significant association was found between TGF-β1 in cancer cells and α-SMA expression in stromal fibroblasts (χ (2) = 0.978, p = 0.326). The 3-year OS rates with low and high TGF-β1 expression in stromal fibroblasts were 52.4 and 26.7 %, respectively (χ (2) = 5.42, p = 0.019). The 3-year OS rates with low and high α-SMA expression in stromal fibroblasts were 53.9 and 31.0 %, respectively (χ (2) =5.01, p=0.025). The multivariate analysis revealed that clinical stage and TGF-β1 and α-SMA expression levels in stromal fibroblasts were identified as independent predictive factors of OS. The results suggest that the expression level of TGF-β1 and α-SMA in stromal fibroblasts may have prognostic significance in patients with clinical stage I-IIIA NSCLC after curative resection.

Immunohistochemical Expression of TGF-β1 in Keloids and Hypertrophic Scars

Aberrant wound healing of skin injury may lead to 2 pathologic entities, termed keloids and hypertrophic scars (HS). There has been growing evidence suggesting a role for transforming growth factor beta (TGF-β) family members in the pathogenesis of fibrosis. The aim of the present work was to investigate the role of TGF-β1 in the pathogenesis of keloids and HS. TGF-β1 was analyzed on skin biopsies of 30 patients presenting with keloids (16) or HS (14) and 10 normal surgical scar and 10 age- and sex-matched normal subjects (controls). TGF-β1 was expressed in dermal fibroblasts, inflammatory cells, and endothelial cells of normal surgical scar (60%) and aberrant scar (86.7%) with an absence of statistical difference. Although it is expressed in 90% of epidermis of aberrant scar (diffuse expression) compared with 60% of normal surgical scar (basal layer expression) and 20% of normal skin biopsies (basal layer expression) with highly significant differences. Dermal TGF-β1 expression in aberrant scar lesions was significantly associated with lesions of shorter duration (P = 0.01) and older age group (P = 0.02). The intense dermal expression was also associated with lesions of shorter duration (P = 0.0001) and immature scars (P = 0.002). TGF-β1 is involved in the pathogenesis of both keloids and HS with maximum effect at early stages. Keratinocytes are not passive partners but rather may have an active role in the induction of fibrosis. Targeting of TGF-β1 may be of benefit if applied early and if directed against keratinocytes as an unusual target of therapy.

Mycophenolate Mofetil Impairs the Integrity of Colonic Anastomosis

The aim of this experimental study was to investigate the effect of mycophenolate mofetil (MMF) during the three phases of colonic anastomosis healing and specifically to check the effect of MMF on the expression of transforming growth factor-beta1 (TGF-beta1), one of the most important growth factors contributing to mechanical stability of colonic anastomosis. Sixty male Wistar rats underwent colonic resection and end-to-end anastomosis. The animals were divided into two groups, a study group given MMF 40 mg/kg, intraduodenally and a control group given vehicle. The rats were sacrificed at 3, 7, and 14 days (10 animals in each group). The anastomoses were tested by measuring bursting pressure and hydroxyproline content. Histological examination and immunohistochemical expression of TGF-beta1 also were assessed. The mean bursting pressure in the study group was significantly lower on day 3 and 7, but there was no statistical significance on day 14. The mean hydroxyproline content was lower in the study group on days 3, 7, and 14. Histology showed decreased number of macrophages and fibroblasts on days 3 and 7 but no difference on day 14. The expression of TGF-beta1 was significantly reduced in the study group, with the difference being more pronounced on days 3 and 7. MMF weakens the integrity of colonic anastomosis, and this effect is more significant during the inflammatory phase of healing. MMF has a negative effect on macrophages and TGF-beta1 expression, resulting in decreased collagen accumulation at the anastomosis.

Expression of Transforming Growth Factor β1 in Bronchial Biopsies in Asthma and COPD

The role of transforming growth factor β1 (TGF β1) in airway remodeling in asthma and chronic obstructive pulmonary disease (COPD) has not been fully described. To evaluate the possible pathogenetic role of TGF β1 in asthma and COPD, immunohistochemical expression of TGF β1 was described in bronchial biopsies from patients with asthma and COPD compared with healthy individuals. Twelve subjects with asthma, 13 subjects with COPD, and 10 healthy individuals enrolled in the study. Bronchial biopsies were stained with hematoxylin and eosin and anti‐TGF β1 antibody. As a result, immunoreactive TGF β1 was mainly localized in association with connective tissue in all groups. The staining intensity was not statistically different among the groups in bronchial epithelium, whereas it was significantly higher in the group of asthma in the submucosa. Because there is evidence showing a significant increase of staining intensity in the submucosa from asthmatics but not from subjects with COPD, we may conclude that TGF β1 may play a significant role in pathogenesis of asthma but not in COPD.

Overexpression of Transforming Growth Factor-β1 in Scirrhous Carcinoma of the Stomach Correlates with Decreased Survival

A retrospective study of 50 patients with scirrhous carcinoma of the stomach who had undergone surgery was performed to elucidate whether immunohistochemical overexpression of transforming growth factor-β1 (TGF-β1) could predict the patients’ survival. To evaluate the proliferative activity of the tumor, the proliferating cell nuclear antigen (PCNA) index was used. Expression of TGF-β1 was low in 27 patients and high in 23. Clinicopathological characteristics were not significantly different between the two groups with high and low TGF-β1 expression. Tumors with higher TGF-β1 expression had higher PCNA indices (p = 0.0467, unpaired Student’s t test). The 5-year survival rate was 40.4% for the low TGF-β1 group and 28.7% for the high TGF-β1 group (p = 0.0388, log rank test). By multivariate analysis, pT, residual tumor, and TGF-β1 were the independent prognostic factors. These results suggest that immunohistochemical expression of TGF-β1 is one of the useful predictors for estimating the patients’ survival in scirrhous carcinoma of the stomach particularly of the T3 and T4 stages.

544 ApolipoproteinE enhances βA4-induced Ca-increases in neurons and astrocytes

We have previously shown that the brains of patients with Alzheimer's disease (AD) express transforming growth factor (TGF)-β2 in neurofibrillary tangle (NFT) -bearing neurons and reactive astrocytes. The present study was undertaken to determine whether other neurodegenerative diseases were also associated with an alteration of the TGF-β's. The immunohistochemical expression of TGF-β1, -2 and -3 was assessed in the brains of patients with progressive supranuclear palsy (n=2), amyotrophic lateral sclerosis (n=3), Lewy body disease (n=5), Parkinson's disease (n=1), Shy-Drager syndrome (n=1), Pick's disease (n=3), lobar atrophy (n=1), and corticobasal degeneration (n=2). Our results were compared to norms for controls (n=8). We found expression of TGF-β2 in both NFT bearing neurons and tangle-bearing glial cells in progressive supranuclear palsy and in neurons with age-related NFT formation. Widespread staining of reactive astrocytes for TGF-β2 was observed in all degenerative diseases. TGF-β1 and -3 staining was not selectively altered in these diseases. We conclude that induction of TGF-β2 may be an intrinsic part of the processes that underlie NFT formation and reactive gliosis in a variety of neurodegenerative diseases.

Transforming Growth Factor-β: Neuronal and Glial Expression in CNS Degenerative Diseases

We have previously shown that the brains of patients with Alzheimer's disease (AD) express transforming growth factor (TGF)-β2 in neurofibrillary tangle (NFT) -bearing neurons and reactive astrocytes. The present study was undertaken to determine whether other neurodegenerative diseases were also associated with an alteration of the TGF-β's. The immunohistochemical expression of TGF-β1, -2 and -3 was assessed in the brains of patients with progressive supranuclear palsy ( n=2), amyotrophic lateral sclerosis ( n=3), Lewy body disease ( n=5), Parkinson's disease ( n=1), Shy-Drager syndrome ( n=1), Pick's disease ( n=3), lobar atrophy ( n=1), and corticobasal degeneration ( n=2). Our results were compared to norms for controls ( n=8). We found expression of TGF-β2 in both NFT bearing neurons and tangle-bearing glial cells in progressive supranuclear palsy and in neurons with age-related NFT formation. Widespread staining of reactive astrocytes for TGF-β2 was observed in all degenerative diseases. TGF-β1 and -3 staining was not selectively altered in these diseases. We conclude that induction of TGF-β2 may be an intrinsic part of the processes that underlie NFT formation and reactive gliosis in a variety of neurodegenerative diseases.