RESUMEN
BACKGROUND: Tinospora cordifolia (Willd).Miers is a perennial climbing medicinal shrub that has been traditionally used for the treatment of chronic inflammatory ailments. Our previous pre- clinical studies on anti-inflammatory effects, proved that the chloroform extract of T. cordifolia (CETC) suppressed the LPS induced up-regulation of pro-inflammatory biomarkers, hence, further follow up study was carried out to evaluate whether CETC can exhibit a protective effect against LPS induced lethal endotoxemia in vivo and also to analyze the impact of CETC pre-treatment on the secretion of pro-inflammatory cytokines in vitro by THP-1 cells. METHODS: To corroborate our previous preclinical studies on inflammation, we investigated the mechanism of the anti-inflammatory effect of T. cordifolia on THP-cells which were pre-incubated with CETC (30 min) and stimulated subsequently with LPS (1 µg/ml) for 20 h. Levels as well as gene expressions of various cytokines were compared with that of LPS alone incubated cells. Alongside, in vivo oral anti-inflammatory efficacy against LPS induced endotoxemia study was effectuated, wherein rats were administered with CETC 48, 24, 12 and 1 h prior to the injection of LPS and the survival of rats were monitored upto 10 days. Cytokine levels were quantified by ELISA. Nitrite levels were measured using Griess reagent. Expression of pro-inflammatory proteins was inspected in rat tissues by histochemical and immuno -histochemical examinations. RESULTS: CETC was able to down-regulate the up-regulation of pro-inflammatory biomarkers in THP-1 macrophages though blockade of NF-κB nuclear translocation and could improve the survival rate during endotoxemic episodes with a marked suppression of the tissue expression of pro-inflammatory proteins. CONCLUSION: These findings concomitantly reveal the anti-inflammatory mechanism of CETC and support us to move forward for the development of drugs against disorders resulting from deregulated immune reactions.
Asunto(s)
Macrófagos/efectos de los fármacos , Extractos Vegetales/farmacología , Sepsis/tratamiento farmacológico , Tinospora/química , Animales , Citocinas/metabolismo , Evaluación Preclínica de Medicamentos , Endotoxemia/tratamiento farmacológico , Humanos , India , Inflamación/tratamiento farmacológico , Lipopolisacáridos , Masculino , FN-kappa B/metabolismo , Ratas , Ratas Wistar , Transducción de Señal/efectos de los fármacos , Células THP-1RESUMEN
AIMS: Superparamagnetic iron oxide nanoparticles (SPIONs), as drug delivery vehicles, offer to eliminate the concerns associated with hydrophobic anti-cancer agents. The current study was intended to fabricate a SPION based delivery system for sorafenib that can simultaneously enable targeted delivery of sorafenib and expand its therapeutic index against hepatocellular carcinoma (HCC). MAIN METHODS: Co-precipitation and physical entrapment methods were employed for the synthesis of sorafenib loaded PVA coated SPIONs. Physicochemical characterizations were done using TEM, XRD, FTIR, Raman spectra and VSM measurements. The superior activity of nanoconjugate was demonstrated by AO/EB staining, FACS, immunofluorescence and Western blot. The safety of the sorafenib conjugated nanoparticles were verified in Wistar rats. KEY FINDINGS: The synthesized nanoparticles were in the size range of 5-15â¯nm. The adsorption of PVA to the SPIONs and the conjugation of sorafenib to the nanocarrier were confirmed by XRD, FTIR and Raman spectra analyses. VSM study ascertained the superparamagnetic nature of the nanoconjugate. Cellular uptake studies suggested its efficient entrapment in HepG2 cells. MTT assay showed that the cytotoxicity of sorafenib loaded PVA/SPIONs was comparable or higher than free sorafenib. The activation of apoptosis and autophagy pathways in HepG2 by the nanoconjugate was evidenced. Acute toxicity testing in Wistar rats supported the safe administration of the nanoconjugate and established its localization in animal tissues by Perl's Prussian Blue reaction. SIGNIFICANCE: The novel combination of sorafenib with PVA/SPIONs showed better anticancer efficiency than free sorafenib demonstrative of its potential in cancer chemotherapy.
Asunto(s)
Antineoplásicos/administración & dosificación , Antineoplásicos/uso terapéutico , Sistemas de Liberación de Medicamentos , Neoplasias Hepáticas Experimentales/tratamiento farmacológico , Neoplasias Hepáticas/tratamiento farmacológico , Nanopartículas de Magnetita , Niacinamida/análogos & derivados , Compuestos de Fenilurea/administración & dosificación , Compuestos de Fenilurea/uso terapéutico , Animales , Antineoplásicos/efectos adversos , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Disponibilidad Biológica , Supervivencia Celular/efectos de los fármacos , Sistemas de Liberación de Medicamentos/efectos adversos , Células Hep G2 , Humanos , Nanopartículas de Magnetita/efectos adversos , Masculino , Proteínas de Neoplasias/biosíntesis , Proteínas de Neoplasias/genética , Niacinamida/administración & dosificación , Niacinamida/efectos adversos , Niacinamida/uso terapéutico , Tamaño de la Partícula , Compuestos de Fenilurea/efectos adversos , Alcohol Polivinílico/química , Ratas , Ratas Wistar , SorafenibRESUMEN
OBJECTIVES: Tinospora cordifolia (Willd.) Miers is an inevitable ingredient of Ayurvedic rasayanas for the treatment of disorders with unregulated inflammation. However, studies regarding the mechanism of anti-inflammatory potential of this plant at the molecular level are lacking. METHODS: In vitro evaluations were conducted in RAW264.7 macrophages which were preincubated with chloroform extract of T. cordifolia (CETC) and subsequently stimulated with LPS. The expressions of COX-2, TNF-α and iNOS genes were analysed by SQRT-PCR and Western blot, cytokines (IL-6, IL-1ß and PGE2 ) levels by ELISA, NF-κB activation and p38 MAPK phosphorylation by Immunoblot and confocal imaging. Anti-inflammatory potential of CETC was validated further in a rat model of carrageenan-induced hind paw edema. Phytochemical characterisation was carried out using the HPLC technique. KEY FINDINGS: The LPS-induced upregulation of proinflammatory biomarkers was significantly prevented by CETC, without inhibiting COX-1. CETC- and LPS-incubated cells showed reduced phosphorylated p38 MAPK levels, and higher levels NF-κB were retained in cytoplasm. Rats pretreated with CETC showed a statistically significant decrease in paw oedema (P ≤ 0.05), and HPLC characterisation detected stigmasterol and ß-sitosterol. The LD50 of CETC lies above 2000 mg/Kg body weight. CONCLUSIONS: These findings encourage us strongly to focus on CETC to develop anti-inflammatory drugs with lower degree of inhibition to the constitutively expressing COX-1.