RESUMO
Although biomechanical changes of the trabecular meshwork (TM) are important to the pathogenesis of glucocorticoids-induced ocular hypertension (GC-OHT), there is a knowledge gap in the underlying molecular mechanisms of the development of it. In this study, we performed intravitreal triamcinolone injection (IVTA) in one eye of 3 rhesus macaques. Following IVTA, we assessed TM stiffness using atomic force microscopy and investigated changes in proteomic and miRNA expression profiles. One of 3 macaques developed GC-OHT with a difference in intraocular pressure of 4.2 mmHg and a stiffer TM with a mean increase in elastic moduli of 0.60 kPa versus the non-injected control eye. In the IVTA-treated eyes, proteins associated with extracellular matrix remodeling, cytoskeletal rearrangement, and mitochondrial oxidoreductation were significantly upregulated. The significantly upregulated miR-29b and downregulated miR-335-5p post-IVTA supported the role of oxidative stress and mitophagy in the GC-mediated biomechanical changes in TM, respectively. The significant upregulation of miR-15/16 cluster post-IVTA may indicate a resultant TM cell apoptosis contributing to the increase in outflow resistance. Despite the small sample size, these results expand our knowledge of GC-mediated responses in the TM and furthermore, may help explain steroid responsiveness in clinical settings.
Assuntos
Glucocorticoides , Pressão Intraocular , Injeções Intravítreas , Macaca mulatta , MicroRNAs , Proteômica , Malha Trabecular , Animais , MicroRNAs/genética , MicroRNAs/metabolismo , Malha Trabecular/efeitos dos fármacos , Malha Trabecular/metabolismo , Glucocorticoides/farmacologia , Glucocorticoides/administração & dosagem , Proteômica/métodos , Pressão Intraocular/efeitos dos fármacos , Pressão Intraocular/fisiologia , Hipertensão Ocular/metabolismo , Triancinolona Acetonida/farmacologia , Fenômenos Biomecânicos , Modelos Animais de Doenças , Microscopia de Força Atômica , Triancinolona/farmacologia , Triancinolona/administração & dosagemRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Euonymus alatus (Thunb.) Siebold. (EA), a traditional Chinese medicine, is widely used in the treatment of diabetes. Our group has previously found that EA could treat diabetic retinopathy (DR) and stigmast-4-en-3-one (Numbered E6) is the active substance responsible for inhibiting angiogenesis in vitro by EA. However, the effects and mechanisms of E6 in the treatment of DR is still unknown. AIM OF THE STUDY: The aim of this study was to investigate the effects and mechanisms of E6 in EA on DR. Additionally, a comparison was made between the effects of E6 and triamcinolone acetonide (TA), as well as the side effects of E6 and dexamethasone. MATERIALS AND METHODS: Ocular affinity assessment and pharmacokinetic parameter prediction were conducted to evaluate the potential of E6 to treat DR. Retinal endothelial cells were used to investigate the in vitro inhibitory effect of E6 on vascular proliferation. Additionally, chicken embryos, zebrafish, and mice were used to investigate the in vivo anti-vascular proliferation effect of E6. Finally, diabetic mice were used to investigate whether E6 improves diabetic retinopathy and to compare its efficacy with that of TA. We then used network pharmacology to study the targets of E6 and performed molecular docking; followed by immunofluorescence experiments, ELISA, Western blot, and tube formation experiments to further investigate its mechanism. Finally, we compared the side effects of E6 with those of dexamethasone. RESULTS: E6 was found to have an affinity for the eye and to inhibit vascular proliferation both in vivo and in vitro. Moreover, E6 was found to be more efficacious than TA in the treatment of DR. Molecular docking experiments predicted that the glucocorticoid receptor (GR) is a potential target of E6, and immunofluorescence analyses confirmed that E6 upregulated the expression of the GR in the retina of hyperglycemic mice. In addition, western blotting results and tube formation experiments showed that E6 also attenuated angiogenesis by inhibiting the Hippo and VEGF pathways. Finally, by comparing the effects of E6 and dexamethasone on glucose regulation and osteoporosis, E6 was found to have fewer side effects. CONCLUSIONS: E6 is a highly effective drug for the treatment of DR, superior to TA and with fewer side effects than dexamethasone. Its mechanism involves the activation of glucocorticoid receptor and inhibition of Hippo and VEGF pathways to alleviate angiogenesis and inflammation. This study is the first to investigate the role and mechanism of E6 in improving DR. The findings suggest that E6 has unique advantages in the treatment of DR.
Assuntos
Diabetes Mellitus Experimental , Retinopatia Diabética , Euonymus , Receptores de Glucocorticoides , Peixe-Zebra , Animais , Retinopatia Diabética/tratamento farmacológico , Camundongos , Receptores de Glucocorticoides/metabolismo , Embrião de Galinha , Diabetes Mellitus Experimental/tratamento farmacológico , Euonymus/química , Masculino , Simulação de Acoplamento Molecular , Dexametasona/farmacologia , Camundongos Endogâmicos C57BL , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Inibidores da Angiogênese/farmacologia , Triancinolona Acetonida/farmacologia , AngiogêneseRESUMO
Purpose: To compare gene expression changes following branch retinal vein occlusion (BRVO) in the pig with and without bevacizumab (BEV) and triamcinolone acetonide (TA). Methods: Photothrombotic BRVOs were created in both eyes of four groups of nine pigs (2, 6, 10, and 20 days). In each group, six pigs received intravitreal injections of BEV in one eye and TA in the fellow eye, with three pigs serving as untreated BRVO controls. Three untreated pigs served as healthy controls. Expression of mRNA of vascular endothelial growth factor (VEGF), glial fibrillary acidic protein (GFAP), dystrophin (DMD), potassium inwardly rectifying channel subfamily J member 10 protein (Kir4.1, KCNJ10), aquaporin-4 (AQP4), stromal cell-derived factor-1α (CXCL12), interleukin-6 (IL6), interleukin-8 (IL8), monocyte chemoattractant protein-1 (CCL2), intercellular adhesion molecule 1 (ICAM1), and heat shock factor 1 (HSF1) were analyzed by quantitative reverse-transcription polymerase chain reaction. Retinal VEGF protein levels were characterized by immunohistochemistry. Results: In untreated eyes, BRVO significantly increased expression of GFAP, IL8, CCL2, ICAM1, HSF1, and AQP4. Expression of VEGF, KCNJ10, and CXCL12 was significantly reduced by 6 days post-BRVO, with expression recovering to healthy control levels by day 20. Treatment with BEV or TA significantly increased VEGF, DMD, and IL6 expression compared with untreated BRVO eyes and suppressed BRVO-induced CCL2 and AQP4 upregulation, as well as recovery of KCNJ10 expression, at 10 to 20 days post-BRVO. Conclusions: Inflammation and cellular osmohomeostasis rather than VEGF suppression appear to play important roles in BRVO-induced retinal neurodegeneration, enhanced in both BEV- and TA-treated retinas. Translational Relevance: Inner retinal neurodegeneration seen in this acute model of BRVO appears to be mediated by inflammation and alterations in osmohomeostasis rather than VEGF inhibition, which may have implications for more specific treatment modalities in the acute phase of BRVO.
Assuntos
Inibidores da Angiogênese , Bevacizumab , Citocinas , Modelos Animais de Doenças , Injeções Intravítreas , Oclusão da Veia Retiniana , Triancinolona Acetonida , Animais , Bevacizumab/farmacologia , Bevacizumab/uso terapêutico , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/administração & dosagem , Triancinolona Acetonida/uso terapêutico , Oclusão da Veia Retiniana/tratamento farmacológico , Oclusão da Veia Retiniana/metabolismo , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/uso terapêutico , Inibidores da Angiogênese/administração & dosagem , Citocinas/metabolismo , Citocinas/genética , Suínos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , Glucocorticoides/farmacologia , Glucocorticoides/uso terapêutico , Regulação da Expressão Gênica/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/metabolismo , Proteína Glial Fibrilar Ácida/genética , Canais de Potássio Corretores do Fluxo de InternalizaçãoRESUMO
We conducted a comprehensive series of molecular biological studies aimed at unraveling the intricate mechanisms underlying the anti-fibrotic effects of triamcinolone acetonide (TA) when used in conjunction with fully covered self-expandable metal stents (FCSEMS) for the management of benign biliary strictures (BBS). To decipher the molecular mechanisms responsible for the anti-fibrotic effects of corticosteroids on gallbladder mucosa, we conducted a comprehensive analysis. This analysis included various methodologies such as immunohistochemistry, ELISA, real-time PCR, and transcriptome analysis, enabling us to examine alterations in factors related to fibrosis and inflammation at both the protein and RNA levels. Overall, our findings revealed a dose-dependent decrease in fibrosisrelated signaling with higher TA concentrations. The 15 mg of steroid treatment (1X) exhibited anti-fibrosis and anti-inflammatory effects after 4 weeks, whereas the 30 mg of steroid treatment (2X) rapidly reduced fibrosis and inflammation within 2 weeks in BBS. Transcriptomic analysis results consistently demonstrated significant downregulation of fibrosis- and inflammation-related pathways and genes in steroid-treated fibroblasts. Use of corticosteroids, specifically TA, together with FCSEMS was effective for the treatment of BBS, ameliorating fibrosis and inflammation. Our molecular biological analysis supports the potential development of steroid-eluted FCSEMS as a therapeutic option for BBS in humans resulting from various surgical procedures. [BMB Reports 2024; 57(4): 200-205].
Assuntos
Fibrose , Inflamação , Triancinolona Acetonida , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Inflamação/tratamento farmacológico , Inflamação/patologia , Humanos , Animais , Constrição Patológica/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Masculino , StentsRESUMO
Contact lenses (CLs) constitute an advantageous platform for the topical release of corticosteroids due to their prolonged contact with the eye. However, the lipophilic nature of corticosteroids hampers CLs' ability to release therapeutic amounts. Two approaches to improve loading and release of triamcinolone acetonide (TA) from poly(2-hydroxyethyl methacrylate)-based hydrogels were investigated: adding 2-hydroxypropyl-ß-cyclodextrin (HP-ß-CD) to the monomers solution before polymerization (HEMA/i-CD) and an hydrogels' post-treatment with HP-ß-CD (HEMA/p-CD). The effect of HP-ß-CD and sterilization by high hydrostatic pressure (HHP) on the hydrogel properties (water content, oxygen and ion permeability, roughness, transmittance, and stiffness) was evaluated. The HEMA/i-CD hydrogels had stronger affinity for TA, sustaining its release for one day. HHP sterilization promoted the formation of cyclodextrin-TA complexes within the hydrogels, improving their drug-loading capacity ¼60 %. Cytotoxicity and irritability tests confirmed the safety of the therapeutic CLs. TA released from the hydrogels permeated through ocular tissues ex vivo and showed anti-inflammatory activity. Finally, a previously validated mathematical model was used to estimate the ability of the TA-loaded CLs to deliver therapeutic drug concentrations to the posterior part of the eye. Overall, HP-ß-CD-containing CLs are promising candidates for the topical ocular application of TA as an alternative delivery system to intraocular injections.
Assuntos
Lentes de Contato Hidrofílicas , Ciclodextrinas , Metacrilatos , Triancinolona Acetonida/farmacologia , 2-Hidroxipropil-beta-Ciclodextrina , Pressão Hidrostática , Corticosteroides , HidrogéisRESUMO
AIM: The primary objective of the research was to develop a cubosomal in situ gel encapsulated with Triamcinolone acetonide (TCA) in order to enhance its penetration through the blood-brain barrier (BBB) when administered via the intranasal route, thus enabling efficient and rapid action. METHOD: Cubosomes were formulated by top-down approach using glyceryl monooleate (GMO), using pluronics127 (PF127) and polyvinyl alcohol (PVA) in varying proportions based on the Box-Behnken design. High resolution transmission electron microscopy (HR-TEM) analysis confirmed the morphology of the cubosomes. The in situ gel was formulated and optimized. Experiments involving ex vivo permeation and histopathology analyses were undertaken to evaluate drug permeation and tissue effects. RESULTS: The cubosomes exhibited a particle size (PS) of 197.9 nm, zeta potential (ZP) of -31.11 mV, and entrapment efficacy (EE) of 84.31%, with low deviation. Batch F4 (19% PF127) showed favorable results. In vitro and ex vivo permeation studies revealed drug release of 78.59% and 76.65%, respectively, after 8 h. Drug release followed the Hixson Crowell model of release kinetics. The histopathological examination revealed no signs of toxicity or adverse effects on the nasal mucosa of the sheep. The formulation exhibited short-term stability, maintaining its integrity and properties when stored at room temperature. CONCLUSION: The utilization of an intranasal cubosomal in situ gel encapsulated with TCA was anticipated to lower intracranial pressure and improve patient adherence by offering effective relief for individuals suffering from Brain edema. This efficacy is attributed to its rapid onset of action and its safe and well-tolerated dosage form.
Assuntos
Portadores de Fármacos , Triancinolona Acetonida , Humanos , Animais , Ovinos , Administração Intranasal , Portadores de Fármacos/farmacologia , Triancinolona Acetonida/farmacologia , Encéfalo , Mucosa Nasal , Tamanho da Partícula , Géis/farmacologiaRESUMO
Traumatic or degenerative joint pain is abundant in the population. Symptom relief by intra- and periarticular glucocorticoid administration is frequently used, however may have potentially devastating effects, changing the normal healing process of the joint. Mesenchymal stem cells (MSCs) are important for wound-healing processes due to their multipotency in regenerating osteoblasts, chondrocytes and adipocytes but also have immunomodulatory properties. The aim of this study was to investigate the impact of triamcinolone acetonide (TA) a common glucocorticoid administrated intra- and periarticularly, on human bone marrow derived MSC viability, functionality, multi-lineage differentiation and transcriptomic output. We found that TA treatment induced apoptosis and promoted adipogenesis while impairing chondrogenesis of MSCs. RNA sequencing indicated that TA modulated the inflammatory response of MSCs, which may have an impact on the immunologic environment where the inflammatory phase is a physiological part of the natural healing process. These data indicate that triamcinolone acetonide should be used with consideration bearing the patient's outcome in mind, with the intention to optimize joint recovery and homeostasis.
Assuntos
Células-Tronco Mesenquimais , Triancinolona Acetonida , Humanos , Triancinolona Acetonida/farmacologia , Glucocorticoides/farmacologia , Diferenciação Celular , Adipogenia/genética , Células da Medula ÓsseaRESUMO
A hypertrophic scar is a complex medical problem. The study of triamcinolone acetonide for the treatment of scars is necessary. The 7mm full-thickness skin wounds were created on the back of BALA/c mice to construct the animal scar model. The different doses of triamcinolone acetonide injection or normal saline were injected into the wound on the 15th, 30th and 45th day after the operation. The skin histopathological changes of mice were observed by Hematoxylin-Eosin (H&E) staining. The proteins and mRNA expression level of scar-biomarkers (COL1, COL3, α-SMA) in mice scar tissue were detected by western blot and qRT-PCR. Besides, the effect of triamcinolone acetonide on the proliferation, invasion, and migration of human hypertrophic scar fibroblast (hHSFs) in vitro was also explored by cck-8, transwell and wound healing assays. After triamcinolone acetonide was injected into the wound, the proportion of scar was significantly reduced, and the treatment effect was concentration-dependently. H&E staining showed that the skin histopathological of mice was improved dose-dependently after injecting the low/middle/high-dosage of triamcinolone acetonide. The proteins and mRNA expression levels of COL1, COL3, and α-SMA were reduced dose-dependently in mice scar tissue. Furthermore, triamcinolone acetonide dose-dependently suppressed the proliferation, invasion, and migration of hHSFs in vitro. Together, triamcinolone acetonide suppressed scar formation in mice and human hypertrophic scar fibroblasts in a dose-dependent manner, phenotypically and mechanistically. The research and further exploration of triamcinolone acetonide in treating scar formation may find new effective treatment methods for the scar.
Assuntos
Cicatriz Hipertrófica , Humanos , Animais , Camundongos , Cicatriz Hipertrófica/tratamento farmacológico , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Pele , Amarelo de Eosina-(YS) , Fibroblastos , RNA Mensageiro/genéticaRESUMO
Triamcinolone acetonide is a glucocorticosteroid used in standard clinical practice for its anti-inflammatory properties. Although it can be given via different routes of administration, the intramuscular route is unique compared with other corticosteroids-its effects remain potent over a longer period of time. We summarize the existing literature on the pharmacokinetic and pharmacodynamic mechanisms of intramuscular triamcinolone acetonide (Kenalog®). The fascinating nature of the purported efficacy of triamcinolone acetonide may be attributed to differing binding mechanisms, low solubility in blood, a low renal clearance rate, and various metabolites and other yet defined effects on skin. The enigma of the purported efficacy of triamcinolone acetonide may lie in the fact that it has a unique nature of having a long-term effect on dermatologic disease using a seemingly low dose compared with other routes of administration and other corticosteroids. Possible reasons for this may be binding differences at the intramuscular site, low solubility due to acetonide esters, a slow rate of absorption from the injected site, and a low renal clearance rate. There is still much to be learned about its mechanism of action, which may be of clinical and therapeutic significance.
Assuntos
Anti-Inflamatórios , Glucocorticoides , Triancinolona Acetonida , Triancinolona Acetonida/administração & dosagem , Triancinolona Acetonida/farmacocinética , Triancinolona Acetonida/farmacologia , Injeções Intramusculares , Humanos , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacocinética , Anti-Inflamatórios/farmacologia , Glucocorticoides/administração & dosagem , Glucocorticoides/farmacocinética , Glucocorticoides/farmacologia , Taxa de Filtração GlomerularRESUMO
Objectives: Keloid is a highly aggressive fibrotic disease resulting from excessive extracellular matrix deposition after dermal injury. Intra-lesional injection of triamcinolone acetonide (TAC) in combination with 5-fluorouracil (5-FU) is a commonly used pharmacological regimen and long-term repeated injections can achieve sustained inhibition of keloid proliferation. However, the molecular mechanisms underlying the inhibitory effect on keloids remain insufficiently investigated. Methods and materials: This study performed single-cell RNA sequencing analysis of keloids treated with TAC+5-FU injections, keloids, and skins to explore patterns of gene expression regulation and cellular reprogramming. Results: The results revealed that TAC+5-FU interrupted the differentiation trajectory of fibroblasts toward pro-fibrotic subtypes and induced keloid atrophy possibly by inhibiting the FGF signaling pathway in intercellular communication. It also stimulated partial fibroblasts to develop the potential for self-replication and multidirectional differentiation, which may be a possible cellular source of keloid recurrence. T cell dynamics demonstrated elevated expression of secretory globulin family members, which may be possible immunotherapeutic targets. Schwann cell populations achieved functional changes by increasing the proportion of apoptotic or senescence-associated cell populations and reducing cell clusters that promote epidermal development and fibroblast proliferation. Conclusions: Our findings elucidated the molecular and cellular reprogramming of keloids by intra-lesional injection of TAC+5-FU, which will provide new insights to understand the mechanism of action and therapeutic targets.
Assuntos
Queloide , Triancinolona Acetonida , Humanos , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Queloide/tratamento farmacológico , Queloide/genética , Queloide/patologia , Fluoruracila , Transcriptoma , Quimioterapia Combinada , Resultado do Tratamento , Injeções IntralesionaisRESUMO
BACKGROUND: The chondrotoxic effects of methylprednisolone acetate (MP) and triamcinolone acetonide (TA) have been well described. However, the mechanical effects of these commonly used steroids on native cartilage are largely unknown. PURPOSE: To investigate the in vitro effects of a single 1-hour MP or TA exposure on the viability, mechanics, and biochemical content of native articular cartilage explants. STUDY DESIGN: Controlled laboratory study. METHODS: Articular cartilage explants (n = 6 per group) were harvested from the femoral condyles of bovine stifles. Explants were exposed to chondrogenic medium containing a clinical dose of MP or TA for 1 hour, followed by fresh medium wash and exchange. Explants in the control group underwent the same treatment with chondrogenic medium alone. At 24 hours after treatment, samples were assessed for viability (live/dead), mechanical properties (creep indentation and Instron tensile testing), biochemical (collagen and glycosaminoglycan) content, and pyridinoline crosslinking via mass spectrometry. RESULTS: Mean cell viability was significantly decreased in native explants exposed to MP (35.5%) compared with the control (49.8%; P < .001) and TA (45.7%; P = .01) specimens. Significant decreases were seen in the mechanical properties of steroid-treated native explants when compared with controls, with decreases in aggregate modulus (646.3 vs 312.8 kPa [MP] and 257.0 kPa [TA]; P < .001), shear modulus (370.1 vs 191.2 kPa [MP] and 157.4 kPa [TA]; P < .001), and ultimate tensile strength (9.650 vs 5.648 MPa [MP; P = .021] and 6.065 MPa [TA; P = .0403]). No significant differences in collagen and glycosaminoglycan content were found in the steroid-treated groups. Pyridinoline crosslinking was significantly decreased in explants exposed to TA compared with controls (P = .027). CONCLUSION: Exposure of MP to articular cartilage explants was chondrotoxic, and exposure of articular cartilage explants to MP or TA resulted in significant decreases in mechanical properties of articular cartilage explants compared with controls. Clinicians should be judicious regarding use of intra-articular steroids, particularly in patients with intact healthy articular cartilage.
Assuntos
Cartilagem Articular , Metilprednisolona , Humanos , Animais , Bovinos , Metilprednisolona/farmacologia , Triancinolona/farmacologia , Triancinolona Acetonida/farmacologia , Acetato de Metilprednisolona , GlicosaminoglicanosRESUMO
OBJECTIVES: Recurrent aphthous stomatitis (RAS) is a painful disorder that commonly appears as ulcers on the oral mucosa, lasting â¼two weeks (minor) to months (major and herpetiform). Current treatment often necessitates the use of topical steroids in the form of pastes, mouthwashes, or gels, but these forms are often ineffective due to inadequate drug contact time with the ulcers. In this study, the performance of novel bilayer mucoadhesive buccal films loaded with triamcinolone acetonide (TA) has been evaluated for targeted drug delivery. METHODS: Experimental mucoadhesive films of hydroxypropyl methylcellulose (HPMC), polyvinyl alcohol (PVA), and polyvinyl pyrrolidone (PVP) were prepared by the solvent casting method, and ethyl cellulose (EC) was applied as the backing layer. The films were characterized for their physical properties, including swelling index (SI), folding endurance, adhesion force with porcine buccal mucosa, residence time and in-vitro drug release. RESULTS: The data showed that the films were flexible with folding endurance> 300 times. With porcine buccal mucosa i) suitable adhesion forces were obtained (between 2.72 and 4.03 N), ii) residence times of> 24 h, and iii) surface pH between 6.8 and 7.1 indicating they would be non-irritant. All films released 100% TA over 6 h, but with varying profiles. The release of TA (over 6 h) from PVP-free films followed Fickian diffusion kinetics (diffusion-controlled release of drug), whereas the mechanism of release from PVP-containing films was found to be a superposition of diffusion-controlled and erosion-controlled release (anomalous). SIGNIFICANCE: The developed films hold great promise for potentially treating RAS and other oral conditions.
Assuntos
Estomatite Aftosa , Triancinolona Acetonida , Animais , Suínos , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Úlcera/tratamento farmacológico , Preparações de Ação Retardada/farmacologia , Preparações de Ação Retardada/uso terapêutico , Estomatite Aftosa/tratamento farmacológico , Mucosa Bucal , Sistemas de Liberação de Medicamentos/métodosRESUMO
BACKGROUND: Tracheobronchial stenosis due to tuberculosis (TSTB) seriously threatens the health of tuberculosis patients. The inflammation and autophagy of fibroblasts affect the development of TSTB. Triamcinolone acetonide (TA) can regulate the autophagy of fibroblasts. Nevertheless, the impact of TA on TSTB and underlying mechanism has remained unclear. OBJECTIVE: To study the impact of TA on TSTB and underlying mechanism. MATERIAL AND METHODS: In order to simulate the TSTB-like model in vitro, WI-38 cells were exposed to Ag85B protein. In addition, the cell counting kit (CCK)-8 assay was applied to assess the function of TA in Ag85B-treated WI-38 cells. Quantitative real-time polymerase chain reaction was applied to detect the mRNA level of sirtuin 1 (SIRT1) and forkhead box O3 (FOXO3a), and autophagy-related proteins were evaluated by Western blot analysis. Vascular endothelial growth factor (VEGF) level was investigated by immunohistochemical staining. Enzyme-linked immunosorbent serologic assay was applied to detect the secretion of inflammatory cytokines. Furthermore, hematoxylin and eosin staining was applied to observe tissue injuries. RESULTS: Ag85B affected WI-38 cell viability in a limited manner, while TA notably suppressed Ag85B-treated WI-38 cell viability. TA induced the apoptosis of Ag85B-treated WI-38 cells in a dose-dependent manner. In addition, Ag85B-treated WI-38 cells demonstrated the upregulation of interleukin (IL)-6, tumor necrosis factor-α (TNF-α), interferon gamma (IFN-γ), and fibrotic proteins (transforming growth factor-beta [TGF-ß] and vascular endothelial growth factor [VEGF]), which can be significantly destroyed by the TA. Meanwhile, TA reversed Ag85-induced inhibition of cell autophagy by mediation of p62, LC3, and Beclin1. Furthermore, silencing of SIRT1/FOXO3a pathway could reverse the effect of TA on the autophagy of Ag85B-treated cells. CONCLUSION: TA significantly induced the autophagy of fibroblasts in Ag85B-treated cells by mediation of SIRT1/FOXO3 pathway. This study established a new theoretical basis for exploring strategies against TSTB.
Assuntos
Sirtuína 1 , Triancinolona Acetonida , Humanos , Triancinolona Acetonida/farmacologia , Sirtuína 1/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/farmacologia , Autofagia , RNA Mensageiro , Proteína Forkhead Box O3RESUMO
Rheumatoid arthritis (RA) is a common systemic autoimmune disease in developed countries. In clinical treatment, steroids have been used as bridging and adjunctive therapy after disease-modifying anti-rheumatic drug administration. However, the severe side effects caused by the nonspecific targeting of organs followed by long-term administration have limited their usage in RA. In this study, poorly water-soluble triamcinolone acetonide (TA), a highly potent corticosteroid for intra-articular injection, is conjugated on hyaluronic acid (HA) for intravenous purposes with increased specific drug accumulation in inflamed parts for RA. Our results demonstrate that the designed HA/TA coupling reaction reveals >98 % conjugation efficiency in the dimethyl sulfoxide/water system, and the resulting HA-TA conjugates show lower osteoblastic apoptosis compared with that in free TA-treated osteoblast-like NIH3T3 cells. Furthermore, in a collagen-antibody-induced arthritis animal study, HA-TA conjugates enhanced the initiative targeting ability to inflame tissue and reduce the histopathological arthritic changes (score = 0). Additionally, the level of bone formation marker P1NP in HA-TA-treated ovariectomized mice (303.6 ± 40.6 pg/mL) is significantly higher than that in the free TA-treated group (143.1 ± 3.9 pg/mL), indicating the potential for osteoporotic reduction using an efficient HA conjugation strategy for the long-term administration of steroids against RA.
Assuntos
Artrite Reumatoide , Triancinolona Acetonida , Camundongos , Animais , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Ácido Hialurônico/farmacologia , Células NIH 3T3 , Artrite Reumatoide/tratamento farmacológico , Injeções Intra-ArticularesRESUMO
In the current work, triamcinolone acetonide (TAA) loaded dual responsive in situ gelling system was designed and optimized using reacted tamarind seed xyloglucan (RXG) (thermoresponsive) and kappa-Carrageenan (κ-CRG) (ion-sensitive) polymers. Tamarind seed xyloglucan (TSX) was subjected to purification followed by enzymatic treatment to produce RXG with ~40 % reduction in galactose content compared to TSX. RXG was characterized using size exclusion chromatography, Fourier transform infrared and proton nuclear magnetic resonance spectroscopy to confirm the ~40 % reduction in galactoside content compared to TSX. The proportions of RXG and κ-CRG in the in situ gels (TAA loaded RXG-κ-CRG) were optimized based on their rheological properties. The optimized in situ gel exhibited good flow properties at 25 °C, but transformed rapidly into a stronger gel in the presence of STF at 35 °C. The optimized formulation had strong mucoadhesion with good spreadability on the surface of excised goat cornea. The drug release followed zero-order kinetics from the optimized in situ gel. Ex vivo ocular toxicity studies indicate that the optimized formulation was well tolerated. The ocular pharmacokinetic studies in rabbits showed significantly higher and sustained vitreous humor exposure of TAA for optimized in situ gel compared to hydroxypropyl-ß-cyclodextrin based aqueous suspension of TAA.
Assuntos
Tamarindus , Triancinolona Acetonida , Animais , Coelhos , Carragenina/química , Triancinolona Acetonida/farmacologia , Géis/química , CórneaRESUMO
Purpose: Apoptosis is a key process in neural degeneration associated with retinal vascular diseases. Vascular endothelial growth factor (VEGF) antagonists, including bevacizumab, are used to treat macular edema in these diseases. As VEGF has a critical role in the preservation of retinal neuronal cells, this study investigates the effects of bevacizumab on neural damage in a pig model of branch retinal vein occlusion (BRVO) and compares it with triamcinolone acetonide (TA) which is reported to possess neuroprotective properties. Methods: Thirty-six pigs had a photothrombotic BRVO in both eyes. Six pigs were injected with bevacizumab in one eye and TA in the fellow eye, then they were sacrificed, the eyes enucleated, and retinas processed at 2, 6, 10, and 20 days, respectively, together with three pigs (six eyes) BRVO only and three normal pigs (six eyes). Neuronal degeneration (apoptosis) and associated inner retinal changes were determined by terminal deoxyynuclotidyl transferase dUTP nick-end labeling (TUNEL), histology, and immunohistochemistry for macrophages. Results: TUNEL labeling showed significantly higher apoptosis rates in the ganglion cell layer (GCL) and the inner nuclear layer (INL) in the bevacizumab-treated compared with the TA-treated retinas at 2, 10, and 20 day time points after occlusion (P < 0.05). Pyknotic cells were significantly higher in the GCL in bevacizumab-treated eyes at 6, 10, and 20 days and in the INL at 2 days compared to TA-treated retinas (P < 0.05). Macrophage infiltration was seen at all time points in both untreated and treated retinas with an absence of significance between bevacizumab- and TA-treated retinas (P > 0.05). Conclusions: Neurodegeneration in the BRVO acute phase is exacerbated by current standard treatments for BRVO. These results may have implications for the timing and treatment type. Translational Relevance: In the acute phase of BRVO, VEGF suppression with bevacizumab and to a lesser extent with triamcinolone exacerbates apoptosis in the inner retinal layers, which has implications for both the timing and choice of treatment.
Assuntos
Oclusão da Veia Retiniana , Triancinolona Acetonida , Suínos , Animais , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Bevacizumab/farmacologia , Bevacizumab/uso terapêutico , Oclusão da Veia Retiniana/tratamento farmacológico , Fator A de Crescimento do Endotélio Vascular , Retina/metabolismoRESUMO
OBJECTIVE: To determine the effect of concurrent versus delayed treatment with corticosteroid on equine articular tissues also treated with local anesthetic in vitro in the presence of inflammatory mediators. STUDY DESIGN: Controlled laboratory study. ANIMALS: Five geldings, one mare (aged 3-18 years). METHODS: From each horse, 24 synovial and 12 osteochondral explants were cultured in a 12-well plate (2 wells/group, 2 synovial and 1 osteochondral explant/well, total 216 explants in the study). Explants were stimulated in culture medium with 10 µg/ml recombinant equine interleukin-1ß and 10 µg/ml tumor necrosis factor-α for 48 hours, then randomly assigned to six treatments: unstimulated control, stimulated control, triamcinolone acetonide (TA, 10-6 M), mepivacaine hydrochloride (MH, 4.4 mg/ml), MH + TA (concurrent) and MH + TA (delayed). The delayed group was treated with MH and, 6 days later, treated with TA. Every 3 days for 9 days total, medium levels of lactate dehydrogenase (LDH), prostaglandin E2 (PGE2 ), matrix metalloproteinase 13 (MMP-13) and glycosaminoglycan (GAG) were quantified via ELISA. Data were analyzed with mixed-effects models with Tukey's multiple comparisons. RESULTS: Stimulation increased medium PGE2 and MMP-13 and had no effect on LDH or GAG. Treatment with MH increased LDH and decreased PGE2 and MMP-13. Treatment with TA decreased PGE2 and MMP-13. CONCLUSION: There were no differences in cytotoxicity, inflammation or matrix degradation for delayed or concurrent MH and TA treatment groups up to 9 days in culture. CLINICAL SIGNIFICANCE: The lack of an effect of concurrent versus delayed treatment might indicate that concurrent therapy is acceptable.
Assuntos
Anestésicos Locais , Cartilagem Articular , Cavalos , Animais , Masculino , Feminino , Anestésicos Locais/farmacologia , Anestésicos Locais/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 13 da Matriz/farmacologia , Corticosteroides/metabolismo , Corticosteroides/farmacologia , Triancinolona Acetonida/metabolismo , Triancinolona Acetonida/farmacologia , Glicosaminoglicanos/análise , Glicosaminoglicanos/metabolismo , Glicosaminoglicanos/farmacologiaRESUMO
Triamcinolone acetonide (TA) has been shown to improve morphological and functional outcome in diabetic macular edema (DME) patients. However, the functional mechanism of TA has not been elucidated yet. In this study we investigated the detailed functional mechanism of TA using culture cells and retinopathy mouse models in which retinal inflammation and abnormal angiogenesis were induced by pericyte depletion. TA significantly prevented retinal hemorrhage, edema and partially improved abnormal angiogenesis. TA decreased retinal vascular endothelial growth factor (VEGF) concentration, presumably by preventing recruitment of macrophages into retina and TA also inhibited expression of inflammatory cytokines in retina. TA inhibited proliferation/migration of vascular endothelial cells and vessel sprouting. No direct inhibition of VEGF receptor 2 (VEGFR2) autophosphorylation was observed by TA. These results suggested that TA improved inflammatory retinal events which were induced in pericyte-deleted mice by mainly decreasing macrophage-derived VEGF and expression of inflammatory cytokines followed by attenuation of vascular permeability and proliferation/migration of endothelial cells. Furthermore, in these processes, translocation of glucocorticoid receptor (GR) was partially involved.
Assuntos
Retinopatia Diabética , Edema Macular , Camundongos , Animais , Triancinolona Acetonida/farmacologia , Triancinolona Acetonida/uso terapêutico , Fator A de Crescimento do Endotélio Vascular , Retinopatia Diabética/tratamento farmacológico , Pericitos , Células Endoteliais/metabolismo , Retina/metabolismo , Inflamação/tratamento farmacológico , CitocinasRESUMO
Standard ocular drug delivery methods generally are safe and effective for treating diseases of the eye. However, many routes of administration carry the risk of adverse effects due to drug exposure to anterior ocular tissues. Additionally, these delivery methods may not result in high and consistent levels of a therapeutic agent delivered to target tissues for diseases affecting the posterior segment of the eye. Injection into the suprachoroidal space (SCS) represents an alternative method of ocular drug delivery to the posterior segment. SCS injection facilitates targeted distribution to affected chorioretinal tissues for potential efficacy benefits, compartmentalization away from unaffected anterior segment tissues for potential safety benefits, and a high degree of bioavailability. Furthermore, the SCS may serve as a drug depot for long-acting drug delivery of small-molecule suspensions. Until recently, drug delivery to the SCS could be achieved only in the operating room setting with anesthetic immobilization of the eye and surgical dissection through the sclera. A novel microneedle device, the SCS Microinjector® (Clearside Biomedical, Inc) was developed to permit physicians to administer therapies safely and reliably into the SCS in the office setting. Successful use of SCS injection has been demonstrated with triamcinolone acetonide injectable suspension (Xipere®, Bausch + Lomb), a novel formulation optimized for use with the SCS Microinjector®. FDA approval of this combination drug and device for the treatment of macular edema associated with uveitis (UME) was based on outcomes from the phase 3 PEACHTREE study (NCT02595398); other important studies included its long-term observational extension (MAGNOLIA; NCT02952001) and an open-label safety study (AZALEA; NCT03097315). The SCS Microinjector® together with triamcinolone acetonide injectable suspension for use in the SCS presents an opportunity for safe and effective drug delivery for the treatment of UME and, potentially, for broader use with alternate medications to treat other ocular diseases that impact chorioretinal tissues (eg, age-related macular degeneration, diabetic retinopathy, choroidal melanoma).
Assuntos
Corioide , Efusões Coroides , Humanos , Microinjeções , Triancinolona Acetonida/farmacologia , Agulhas , Estudos Observacionais como AssuntoRESUMO
The four extraocular rectus muscles in the rabbits were disinserted for induction of anterior segment ischemia (ASI) and the changes in the concentrations of prostaglandin E2 (PGE2), hypoxia-inducible factor-1 (HIF-1α), and vascular endothelial growth factor (VEGF) in the aqueous and vitreous humor were evaluated. Disinsertion of four rectus muscles in rabbits was performed in the right eyes of rabbits (ASI group). The concentrations of PGE2, HIF-1α, and VEGF in the aqueous and vitreous humor were measured at 1, 3, 6, 12, and 24 h by ELISA. The concentrations were compared with those of the fellow eyes (contralateral group) and normal healthy eyes (control group). Subconjunctival injection of triamcinolone acetonide (TA) was administered and three cytokine concentrations in the aqueous humor and vitreous humor were measured at 12 h after TA injection. A total of 48 eyes from 28 rabbits were included. The concentrations of PGE2, HIF-1α, and VEGF in the aqueous humor in the ASI and contralateral groups were significantly higher than those in the control group (p < 0.05, all). The aqueous and vitreous humor concentrations of VEGF in eyes with simultaneous TA injection were significantly lower than were those in the ASI group (p = 0.02, all). The concentration of PGE2, HIF-1α, and VEGF in the aqueous humor was increased after induction of ASI and TA injection seems to be effective in inhibiting VEGF elevation in ASI.