Effects of tranilast on the epithelial-to-mesenchymal transition in peritoneal mesothelial cells

BACKGROUND: We investigated the effects of tranilast on epithelial-to-mesenchymal transition (EMT) in an animal model and on the EMT signaling pathway in human peritoneal mesothelial cells (HPMCs).METHODS: We performed in vitro studies (cytotoxicity, cell morphology, and western blot analyses) on HPMCs from human omenta, along with in vivo studies (peritoneal membrane function and morphometric and immunohistochemical analyses) on Sprague Dawley rats. Thirty-two rats were divided into three groups: control (C) group (peritoneal dialysis [PD] catheter but not infused with dialysate), PD group (4.25% glucose-containing dialysate), and PD + tranilast group (4.25% glucose-containing dialysate along with tranilast).RESULTS: In in vitro experiments, transforming growth factor-beta 1 (TGF-β1) increased α-smooth muscle actin and Snail expression and reduced E-cadherin expression in HPMCs. TGF-β1 also reduced cell contact, induced a fibroblastoid morphology, and increased phosphorylation of Akt, Smad2, and Smad3 in HPMCs. Tranilast significantly inhibited TGF-β1-induced EMT and attenuated these morphological changes in HPMCs. In in vivo studies, after 6 weeks of experimental PD, the peritoneal membrane was significantly thicker in the PD group than in the C group. Tranilast protected against PD-induced glucose mass transfer change and histopathological changes in rats.CONCLUSION: Tranilast prevented EMT both in HPMCs triggered with TGF-β1 and in rats with PD-induced peritoneal fibrosis. Thus, tranilast may be considered a therapeutic intervention that enables long-term PD by regulating TGF-β1 signaling pathways.

A Case of Vulvar Milium-like Syringoma in a Child Treated with Tranilast / 대한피부과학회지

Syringoma is the most common type of benign intraepidermal eccrine sweat gland tumor in Korea, and is usually found in women in their forties. It presents mostly as a localized lesion, preferring the lower eyelid, cheek, or forehead, and rarely invades the vulval area, and in the case of children, vulvar invasion is even more rare. Tranilast is an antihistamine used for atopic dermatitis and asthma, and has recently been used for the treatment of keloid. A few previous studies have reported both localized and generalized forms of syringoma being effectively resolved with tranilast. Herein, we report a rare and interesting case of milium-like syringoma, which manifested on the vulval area of 10-year old girl that was successfully treated with tranilast.

Inhibition Effects of Tranilast on Hypertrophic Scar Tissue of Rabbits and Its Mechanism Study / 中国药房

OBJECTIVE:To study the inhibition effects of tranilast on hypertrophic scar tissue of rabbits and its mechanism. METHODS:Rabbits were selected to induce hypertrophic scar(HS)model,the HS model rats were randomly divided into model control group (normal saline),tranilast low-dose,medium-dose,high-dose groups (0.3,0.5,0.7 mg/kg),6 rabbits in each group,local intradermaly injected corresponding drugs. Scar thickness 1 h before injection and the first,third,fifth week after in-jection in each group were measured,pathological changes of scar(fifth week after injection)were observed,transforming growth factor β1 (TGF-β1),α-smooth muscle actin(α-SMA)mRNA and protein expression were detected. RESULTS:Compared with 1 h before injection,scar thickness of rabbits in other groups were decreased after injection except for model control group. In fifth week after injection,compared with model control group,scar thickness of rabbits in other groups were decreased,pathological changes were improved;TGF-β1,α-SMA mRNA and protein expression were decreased (P<0.05),showing positive correlation with tranilast dose. CONCLUSIONS:Tranilast can inhibit the formation of hypertrophic scar,and the mechanism may be related to inhibiting the TGF-β1,α-SMA mRNA and protein expression.

Effect of tranilast on cardiac fibroblasts proliferation and phenotype transformation incubated with high concentration of glucose / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of tranilast on cardiac fibroblasts proliferation and phenotype transformation incubated with a high concentration of glucose and the possible mechanism. METHODS The cardiac fibroblasts were divided into seven groups in accordance with different nutrient solutions:normal control group(5.5 mmol · L-1 glucose),hypertonic group(5.5 mmol · L-1 glucose+mannitol 25 mmol · L- 1),high glucose group (25 mmol · L- 1 glucose),tranilast intervention groups (25 mmol·L-1 glucose+tranilast 50,100 and 200μmol·L-1),and activin receptor-like kinase 7(ALK7) inhibitor group(25 mmol · L-1 glucose+10μmol · L-1 SB431542). The cell proliferation was detected by MTT method. The transformation of cardiac fibroblasts was determined by immunfluorescence staining. The expression of fibroblast specific protein 1 (FSP-1),α-smooth muscle actin (α-SMA),transforming growth factor-β1(TGF-β1)and ALK7 was assessed by Western blotting. RESULTS Compared with nor?mal control group,A492 nm of cells in high glucose group was significantly increased(P<0.01),while the expression of α-SMA,TGF-β1 and ALK7 protein in high glucose group was significantly increased(P<0.01),but FSP-1 protein was significantly decreased(P<0.01). There was no difference between normal control group and hypertonic group. Compared with high glucose group,A492 nm of cells in tranilast or SB431542 intervention groups were decreased(P<0.05),and the expression of α-SMA,TGF-β1 and ALK7 protein was significantly decreased(P<0.05),but the expression of FSP-1 protein was increased (P<0.05)in tranilast or SB431542 intervention groups. CONCLUSION Tranilast can inhibit the proliferation and phenotype transformation of cardiac fibroblasts induced by high glucose,which may be related to down-regulation of the expression of ALK7.

The Result of Photorefractive Keratectomy Treated with 0.1% Fluorometholone and Tranilast Eye Drops

PURPOSE: To determine the effectiveness of the method for preventing corneal opacity and minimizing the intraocular pressure (IOP) increase after photorefractive keratectomy treated with 0.1% fluorometholone and tranilast (0.5% tranilast, Krix®, JW pharmaceutical, Seoul, Korea), especially in cases with elevated IOP. METHODS: The patients who underwent photorefractive keratectomy from May 2014 to May 2015 were enrolled in the present study. The data of 49 patients (49 eyes) with elevated IOP at 1 month postoperatively and who used 0.1% fluorometholone and tranilast eye drops (tranilast group) were analyzed and compared with the control group consisting of patients who underwent the same surgery from December 2012 to October 2013 but used only 0.1% fluorometholone. RESULTS: The visual acuity at postoperative 6 months was log MAR -0.08 ± 0.05 and log MAR -0.08 ± 0.04 in the tranilast group and control group, respectively. The eye drops were used postoperatively for 17.7 ± 3.3 weeks in the tranilast group and for 20.5 ± 3.7 weeks in the control group (p < 0.01). Anti-glaucoma eye drops were used for 18.4 ± 3.2 weeks and 20.9 ± 3.7 weeks postoperatively in the tranilast group and control group, respectively (p < 0.01). CONCLUSIONS: Adding tranilast eye drops to patients whose IOP was elevated because of 0.1% fluorometholone use after photorefractive keratectomy is an effective method for preventing corneal haze and minimizing IOP elevation.

Evaluation of the Combination of Methylprednisolone and Tranilast after Spinal Cord Injury in Rat Models

OBJECTIVE: The aim of our study was to evaluate the neuroprotective functions of the combination therapy using methylprednisolone (MP) and tranilast (TR) after spinal cord injury (SCI) in adult rats. METHODS: Spinal cord compression injury model was achieved using Yasargil aneurysm clip. Rats were divided into control group, MP group, TR group, and combination therapy group using TR and MP. Rat models were assessed for locomotor functional recovery using Basso, Beattie, and Bresnahan (BBB) score, spinal cord water content and myeloperoxidase (MPO) activity 24 hours post SCI, haematoxylin and eosin staining and glial fibrillary acid protein (GFAP) staining at 7 and 14 days post SCI. RESULTS: The spinal cord water content and MPO activity in the combination therapy group was significantly lower than the control group and the individual therapy groups p0.05). At 2 weeks after SCI there was a slight decrease in GFAP expression compared to the first week but the difference was not statistically significant (p>0.05), GFAP expression between the groups was not statistically significant p>0.05. CONCLUSION: Combining MP and TR is therapeutically more effective in improving functional recovery, inhibiting inflammation and glial scar formation after acute SCI.

Comparison of Effect on Antioxidative Stress Between Benezepril and Tranilast in Diabetic Nephropathy Rat / 医药导报

Objective To investigate ACEI ( benazepril ) and tranilast exert renoprotective properties in diabetic nephropathy( DN) through the inhibition of thioredoxin( Trx) . Methods Forty male SD rats were randomly divided into normal control group,model control group,tranilast group and benazepril group (n=10 each).Normal control group was fed with normal diet. Other groups were fed with high-glucose high-fat diet to make DN models. Rats in model control, tranilast, and benazepril groups were fed with normal diet,400 mg??kg-1??d-1 tranilast plus normal diet,and 10 mg??kg-1??d-1 benazepril plus normal diet,respectively,via oral gavage for 12 weeks.The 24-hour proteinuria,blood glucose(BG),blood urea nitrogen (BUN),serum creatinine ( Scr) and renal pathology changes were detected. Expression of Trx was measured by Western-blot. Results The 24 h urine protein, BG, BUN, Scr, kidney/body weight, and glomerular sclerosis index were significantly decreased in tranilast group and benazepril group,as compaired with model control group ( P0.05).Both tranilast and benazepril can reduce renal pathological changes,and can increase the expression of Trx of DN rats, but benazepril had a more significant effect on increasing Trx expression. Conclusion Both tranilast and benazepril have renoprotective function in DN, and benazepril is more effective than tranilast in delaying the progression of diabetic nephropathy by increasing Trx expression and decreasomg oxidative stress.

TRANILAST ADMINISTRATION TIME FOR SUPPRESSION OF BURN SCAR HYPERPLASY / 现代医院

Objective To study the clinical effects of oral administration of tranilast to suppress burn scar hyperplasy when the burn wounds began repairing or were completely healed.Methods Sixty burn patients from July 2011 to December 2013 were randomly divided into groups A and B, 30 patients in each group.Thirty patients were treated with orally-administered tranilast at the beginning of wound repair, while the other patients were treated with orally administered tranilast when the burn wounds were completely healed.The therapeutic effect of these two groups was compared.Results Six months after burn wound healing, Vancouver scar scale was applied to evaluate the ther-apeutic effect.No significant differences were found between these two groups ( p>0.05) , and no adverse reactions were observed.Conclusion No significant differences in clinical effects of oral tranilast administered at the begin-ning of burn wound repair and at the time point of complete healing.

Effects of Topical Tranilast on Corneal Haze with the Pentacam(R) after Photorefractive Keratectomy

PURPOSE: To evaluate the inhibitory effect of tranilast on formation of corneal haze using the Pentacam(R) after photorefractive keratectomy (PRK). METHODS: A prospective, randomized, paired eye study was performed. A total of 60 eyes from 30 patients were enrolled in the present study. Eyes were categorized as myopic eyes or =-5 D. Patients undergoing PRK were randomized to receive tranilast in one eye and no medication in the contralateral eye. Three months postoperatively, corneal haze was measured with the Pentacam(R) and compared between the 2 groups. RESULTS: Statistical differences were not found in preoperative data in the tranilast or control groups (all P > 0.05). There was a strong decreasing density trend from the apex to the 3 mm radius in both groups (P 0.05). CONCLUSIONS: The use of tranilast after PRK did not inhibit corneal opacity. Additionally, Pentacam(R) can provide a useful objective measure of corneal haze.

Role and mechanism of tranilast preventing the progression of tubulointerstilial ifbrosis in diabetic kidney diseases / 中南大学学报(医学版)

Objective:To determine the role and mechanism of tranilast preventing the progression of tubulointerstilial ifbrosis in diabetic kidney disease (DKD). Methods:Sprague-Dawley rats were randomly divided into a control group (n=6), DKD model group (n=8), low dose tranilast group [200 mg/(kg.d), n=8], and high dose tranilast group [400 mg/(kg.d), n=8]. Tranilast was administered daily after the model was built. Rats were sacrificed at day 56, 24 hour urine was collected to measure 24-hour urine albumin excretion, and blood was collected to determine the renal function and serum albumin. Then the kidneys were harvested and subjected to studies. The expression of C3aR, E-cadherin,α-SMA, fibronectin(FN), collagen I (Col I), stem cell factor (SCF) and c-kit were detected by immunohistochemical staining respectively. The expression of E-cadherin,α-SMA, FN, Col I, SCF and c-kit protein was analyzed by Western blot, and the expression of FN, Col I, SCF and c-kit mRNA was examined by RT-PCR. Results:Tranilast can inhibit the inifltration of mast cells in the kidneys of DKD rats. The expression ofα-SMA in the kidneys of DKD rats inereased signiifcantly (P Conclusion:Mast cells participate in and aggravate the renal tubulointerstitial fibrosis in DKD rats. Tranilast can reverse the EMT of renal tubular cells and inhibit the tubulointersitial fibrosis of DKD by blocking the inifltration of mast cells induced by SCF/c-kit pathway.

Influence of tranilast on the cyclosporine A-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells / 中华器官移植杂志

Objective To study the effect of tranilast on cyclosporine A (CsA)-induced epithelial-to-mesenchymal transition in human renal tubular epithelial cells, and investigate the mechanism of its antifibrotic effect. Methods Cultured HK-2 cells were divided into four groups: (1)In the control group, cells were treated without any medicine; (2) The cell were treated with CsA (4. 2μmol/L) for 72 h; (3) The cells were treated with a combination of CsA (4. 2 μmol/L) and tranilast (100μmol/L); (4) The cells were treated with tranilast (100 μmol/L) alone for 72 h.Morphological changes of the cells were assessed by phase-contrast microscopy. The immunofluorescence and Western blotting were adopted to detect the expression of E-cadherin, α-SMA and OPN mRNA and proteins respectively. Results Tranilast could markedly ameliorate the morphological changes of HK-2 cells stimulated by CsA. The irmmunofluorescence staining revealed the expression of E-cadherin was markedly decreased in HK-2 cells stimulated with CsA for 72 as compared with the control group, while the expression of α-SMA and OPN was significantly higher in CsA group than the control group. The expression of E-cadherin in the CsA + Tranilast group was higher than the CsA group, while the expression of α-SMA and OPN in the CsA + Tranilast group was lower than the CsA group. Western blotting showed that protein expression level of E-cadherin in CsA group was dramatically lower than that in the control group (P＜0. 05), while that of α-SMA and OPN in CsA group was significantly higher than in the control group (P＜0.05). The protein expression level of E-cadherin in HK-2 cells in the CsA + Tranilast group was markedly higher than in the CsA group (P＜0.05), and that of α-SMA and OPN in CsA + Tranilast group was significantly lower than in the CsA group (P＜0. 05). Conclusion Tranilast can block the CsA-induced epithelialto-mesenchymal transition in HK-2 cells probably by suppressing the expression of OPN.

The Effectiveness of Tranilast in the Prevention of Posterior Capsular Opacity

PURPOSE: To evaluate the inhibitory effect of tranilast on the formation of posterior capsular opacity (PCO) after a cataract operation ex vivo and in a rabbit model. METHODS: A human lens epithelial cell line (B3) was treated with 0.005-0.1 mM tranilast. Cytotoxicity assessment and effective dosage determination of tranilast were performed using MTT assays. B3 cell lines were cultured in Eagle's minimal essential medium (EMEM) containing 20% FBS with different concentrationsof tranilast, and morphological differences were observed. To investigate the effect of tranilast on cytokine production in B3 cell lines, cells were treated with 0.01 mM tranilast and expression profiles of cytokines were analyzed by RT-PCR. After performing phacoemulsification and intraocular lens implantation in 10 white rabbits, 0.5% tranilast eye drops were given 4 times per day, and the severity of PCO was evaluated bi-weekly using POCOman for 8 weeks after the operation. RESULTS: Cell death was observed in the 0.05 mM tranilast-treated B3 cell lines, and inhibition of epithelial-mesenchymal transition (EMT) was also observed in the 0.01 mM tranilast-treated B3 cell lines. TGF-beta1/2, IL-18, and CDK7 mRNA expression decreased in the 0.01 mM tranilast-treated B3 cell lines. Significant suppression of PCO formation was observed in rabbits treated with 0.5% tranilast eye drops 5 weeks post operative (p<0.05). CONCLUSIONS: The results from this study show that tranilast suppresses EMT through inhibition of TGF-beta, IL-18,and CDK7 expression. The results suggest that tranilast can be used toprevent PCO formation after cataract surgery.

The Effectiveness of Tranilast in the Prevention of Posterior Capsular Opacity

PURPOSE: To evaluate the inhibitory effect of tranilast on the formation of posterior capsular opacity (PCO) after a cataract operation ex vivo and in a rabbit model. METHODS: A human lens epithelial cell line (B3) was treated with 0.005-0.1 mM tranilast. Cytotoxicity assessment and effective dosage determination of tranilast were performed using MTT assays. B3 cell lines were cultured in Eagle's minimal essential medium (EMEM) containing 20% FBS with different concentrationsof tranilast, and morphological differences were observed. To investigate the effect of tranilast on cytokine production in B3 cell lines, cells were treated with 0.01 mM tranilast and expression profiles of cytokines were analyzed by RT-PCR. After performing phacoemulsification and intraocular lens implantation in 10 white rabbits, 0.5% tranilast eye drops were given 4 times per day, and the severity of PCO was evaluated bi-weekly using POCOman for 8 weeks after the operation. RESULTS: Cell death was observed in the 0.05 mM tranilast-treated B3 cell lines, and inhibition of epithelial-mesenchymal transition (EMT) was also observed in the 0.01 mM tranilast-treated B3 cell lines. TGF-beta1/2, IL-18, and CDK7 mRNA expression decreased in the 0.01 mM tranilast-treated B3 cell lines. Significant suppression of PCO formation was observed in rabbits treated with 0.5% tranilast eye drops 5 weeks post operative (p<0.05). CONCLUSIONS: The results from this study show that tranilast suppresses EMT through inhibition of TGF-beta, IL-18,and CDK7 expression. The results suggest that tranilast can be used toprevent PCO formation after cataract surgery.

Influence of tranilast on expression of OPN and interstitial ED1+ macrophages infiltration in rats with chronic cyclosporine A nephrotoxicity / 中国实用内科杂志

Objective To evaluate the effect of tranilast on chronic cyclosporin A(CsA)nephrotoxicity rats and the possible mechanisms of protective effect of it.Methods Rats were on low salt diet and CsA was administered by gastric gavage at a dose of 20 mg/(kg?d)for 28 days,and tranilast was gave to these rats in a dose of 400 mg/(kg?d).The renal function and renal morphology were observed at1,2 and 4 weeks after administration of tranilast.Results CsA induced the corruption of normal condition and weight lost were markedly delayed after administration of tranilast.On histopathology tranilast attenuated tubular cells vacuolations as well as interstitial damage including interstitial ED1+ macrophages infiltration and focal interstitial fibrosis.Meanwhile,tranilast down-regulated the protein of OPN in kidney.Conclusion Tranilast can attenuate the structural changes of kidney in chronic CsA nephrotoxicity rats.Decreased CsA-related ED1+ macrophages,OPN up-regulation expression in the kidney may be related to mechanisms of protective effect of tranilast in the treatment of CsA induced chronic nephrotoxicity.

Effect of Tranilast on the Proliferation of Human Corneal Keratocytes in Vitro

PURPOSE: To evaluate the inhibitory effect of tranilast on proliferation of cultured human keratocytes, and to investigate the apoptotic response and the cellular morphologic changes associated with tranilast in vitro. METHODS: Human corneal keratocytes were exposed to tranilast at a concentration of 0.05, 0.1, 0.2, 0.4, 0.8, 1.6, and 3.2 mg/ml for a period of 4, 24, and 48 hours. Evaluations were conducted with MTT-based-calorimetric assay for measuring the metabolic activity, flow cytometric analysis and fluorescent micrograph for assessing the apoptotic response, and inverted phase-contrast micrograph and electron microscopy for observing the morphologic changes. RESULTS: The inhibitory effect of human keratocyte proliferation was found to have a dose and time dependent pattern (p<0.05). In flow cytometry, the maximal apoptotic response developed at 0.8 mg/ml concentration after 4 and 24 hours of exposure time, and apoptotic cells were demonstrated in fluorescent micrograph. At higher concentration of Tratnilast, human corneal keratocytes were more swollen rather than having a spindle shape and being detached from the bottom of the dish. The damaged keratocytes had degenerative and apoptotic changes like the formation of phagolysosomal granule, marginal condensation in the nucleus, and bleb formation of the nuclear membrane. CONCLUSIONS: The apoptotic response of tranilast is concerned with the inhibitory effect of human corneal keratocyte proliferation. Therefore, tranilast shows promise in clinical use for the inhibition of postoperative excimer laser induced corneal opacity or haze with fewer side effects.

The clinical effects of probucol combined with multivitamins and Tranilast on restenosis after percutaneous coronary intervention / 대한내과학회지

BACKGROUND: Tranilast is an anti-allergic drug that suppresses the release of cytokines. An antioxidant, probucol, prevents endothelial dysfunction and oxidation of low density lipoprotein and also inhibits the secretion of interleukin-1 by macrophages. In several studies, both the tranilast and probucol with multivitamins have been shown to decrease the frequency of angiographic restenosis after PCI. METHODS: We analyzed clinical events and restenosis at 6 months following percutaneous coronary angioplasty in 93 patients with 113 coronary arterial lesions after coronary stenting at Soonchunhyang University Hospital between Jan 2001 and Apr 2003. The patients were assigned to following three groups: 39 patients who didn't receive tranilast and antioxidants (control group, M 29, F 10, 61 +/- 10 years) ; 25 patients who received probucol (500 mg), vitamin C (1,000 mg), vitamin E (400 mg) (antioxidant group, M 19, F 6, 62 +/- 10 years) ; 29 patients who received tranilast (400 mg) (Tranilast group, M 18, F 11, 59 +/- 9 years). RESULTS: The restenosis per lesion between three groups was not different significantly (control group, 32.7%; antioxidant group, 26.7%; Tranilast group, 20.6%). At follow-up, minimal luminal diameter (MLD) was not different significantly between three groups (control group, 1.8 +/- 1.07 mm; antioxidant group, 2.1 +/- 1.18 mm; Tranilast group, 2.1 +/- 0.94 mm). Target lesion revascularization was lower in Tranilast group (3.4%) as compared with control group (25.6%) and antioxidant group (16%, p<0.05). CONCLUSION: Neither probucol combined with vitamin C and E nor tranilast did not improve significantly the angiographic restenosis rate. But tranilast had reduced the target lesion revascularization rate as compared with control group and antioxidant group.

Effect of tranilast on airway remodeling of experimental COPD rats / 中国临床药理学与治疗学

AIM: To observe the effect of tranilast on experimental COPD rats in terms of airway remodeling. METHODS: Forty eight SD rats were divided into two groups in random: untreated model group, tranilast-treated group. Another eight rats were selected as control group. The COPD rat model was established by passive smoking and intratracheal instillation of lipopolysaccharide (LPS) and then treated with sterile saline or tranilast (400 mg?kg~ -1 ?d~ -1 ) respectively. Eight rats in each group were killed in 7th, 14th, 28th day after the beginning of proceeding. Bronchoalveolar lavage fluid (BALF) was collected, and the total and differential cells were counted. The distribution and the ratio of type I to type III collagen in the lung tissue were determined using a sirius red polarizing microscopy morphometry method. Lung tissues were observed by hemotoxylin and eosin stain, then the image analysis were made. RESULTS: The total cells and the AM ratio in BALF of tranilast-treated group significantly decreased in comparison with those in model group (P

The Effects of Tranilast gamma on the Cellular Proliferation and alpha-Smooth Muscle Actin Expression in the Cultured Keratocytes of Rabbit

PURPOSE: To evaluate the effects of Tranilast gamma on the cellular proliferation and expression of alpha-smooth muscle actin on the cultured keratocytes of the rabbit that influence the formation of corneal scar and haze in vitro. METHODS: After the keratocytes of the rabbit were cultured, they were exposed to Tranilast gamma (N-(3, 4-dimethoxycinnamoyl) anthranilic acid) 0.05, 0.1, 0.2, 0.4, 0.8, 1.0, 1.6 mg/ml with DMEM used as a control. MTT was used to measure the metabolic activity, and the Western blot assay was performed to confirm the alpha-smooth muscle actin expression, which was expressed with time by Tranilast gamma treatment. RESULTS: Higher the concentration and longer the exposure time of Tranilast gamma, the more the inhibition of the cellular proliferation. LD50 concentration was about 0.4 mg/ml in the exposure time of 24 hours and 0.2 mg/ml in the exposure time of over 48 hours(P<0.05). There was a decrease in response of alpha-smooth muscle actin expression with increasing concentrations of Tranilast gamma relative to the control. CONCLUSIONS: Tranilast gamma trends to have an inhibitory effect on the cellular proliferation and induction of alpha-smooth muscle actin expression. Tranilast gamma may inhibit excessive haze of the cornea and inhibit scar formation after the corneal wound healing and photorefractive surgery in the cornea.

The Effects of Tranilast(R) on the Cellular Proliferation, Collagen Synthesis, and Secretion of TGF-beta1 in Bovine Retinal Pigment Epithelial Cells

PURPOSE: To evaluate the effects of Tranilast(R) on the cellular proliferation, collagen synthesis, and secretion of transforming growth factor-beta1 on retinal pigment epithelial cells that influence the development of proliferative vitreoretinopathy in vitro. METHODS: After bovine RPE cells were isolated, they were exposed to Tranilast(R) 0.05, 0.1, 0.2, 0.4, 0.8, 1.0, 1.6 mg/ml and only DMEM which was used as control. MTT was used as a measurement of metabolic activity, and collagen synthesis and TGF-beta1 secretion were assayed by collagen kit (Sigma, USA) and TGF-beta1 kit (Gibco, USA). Western blot assay was used to confirm TGF-beta1, which was expressed by treated Tranilast(R) in bovine RPE cells. RESULTS: The higher the concentration of Tranilast(R), the more the inhibition of the cellular proliferation. LD50 concentration was about 1.0 mg/ml, and there was more significant difference the concentration of over Tranilast(R) 0.4 mg/ml than control (P<0.05). The higher the concentration of Tranilast(R), the more the inhibition of collagen synthesis and TGF-beta1 secretion from RPE cells, especially over Tranilast(R) 0.05 mg/ml (P<0.05). There was no response of TGF-beta1 expression in the concentration of Tranilast(R) 0.2 mg/ml compared to untreated bovine retinal epithelial cells. CONCLUSIONS: Tranilast(R) has a tendency of inhibitory effect in the cellular proliferation, collagen synthesis, and TGF-beta1 secretion. Tranilast(R) may prevent excessive proliferation of RPE and scar formation concerned with collagen synthesis and TGF-beta1 secretion in the proliferative vitreoretinal diseases.

Inhibitory Effect of Tranilast on the Proliferation of Human Conjunctival Epithelial Cells and Subconjunctival Fibroblasts

PURPOSE: Tanilast has been clinically used for various allergic diseases. Recently, it has also been found to inhibit excessive scarring in wound healing process. The purpose of this study is to investigate the inhibitory effect of Tranilast on the proliferation of human conjunctival epithelial cells and subconjunctival fibroblasts. METHODS: Human conjunctival epithelial cells and subconjunctival fibroblasts were exposed for 24 hours to Tranilast 0.05, 0.2, 0.4, 0.8, and 1.6 mg/ml. MTT based calorimetric assay was performed to assess the metabolic activity and inhibition of cellular proliferation. RESULTS: As the concentration of Tranilast increased, the absorbance of spectrometer decreased. Inhibitory effect of cellular proliferation was stronger in subconjunctival fibroblasts than in conjunctival epithelial cells. The inhibitory effect of celluar proliferation in conjunctival epithelial cells was 75%, 66%, 64%, 59%, and 39% at 0.05, 0.2, 0.4, 0.8, and 1.6 mg/ml, respectively (LD50 was 0.9 mg/ml). In subconjunctival fibroblasts, the inhibitory effect of celluar proliferation was 66%, 62%, 47%, 33%, and 23% at 0.05, 0.2, 0.4, 0.8, and 1.6 mg/ml, respectively (LD50 was 0.3 mg/ml). CONCLUSION: Tranilast may prevent excessive proliferation of both human conjunctival epithelial cells and subconjunctival fibroblasts. This inhibitory effect on cellular proliferation was higher in subconjunctival fibroblasts than in conjunctival epithelial cells, suggesting the possibility of being used after glaucoma filtering surgery or pterygium excision.