RESUMEN
Protocatechuic acid (PCA), chlorogenic acid (CA) and luteolin (LT) are plant phenols found in Chinese medicinal herbs such as Lonicera japonica. Cytotoxicity assays showed that PCA, CA and LT (at 100 micromol/L) effectively killed the HepG2 hepatocellular carcinoma cells. Among these three naturally occurring compounds, only PCA was capable of stimulating the c-Jun N-terminal kinase (JNK) and p38 subgroups of the mitogen-activated protein kinase (MAPK) family. Coincidently, PCA-induced cell death was rescued by specific inhibitors for JNK and p38, while the cytotoxicities of CA and LT were partially eliminated by the antioxidant effect of N-acetyl-L-cysteine (NAC). Further investigation demonstrated that the aqueous extract of Lonicera japonica also triggered HepG2 cell death in a JNK-dependent manner, but the amount of PCA alone in this herbal extract was insufficient to contribute the subsequent cytotoxic effect. Collectively, our results suggest that PCA is a naturally occurring compound capable of inducing JNK-dependent hepatocellular carcinoma cell death.
Asunto(s)
Antineoplásicos/farmacología , Carcinoma Hepatocelular/tratamiento farmacológico , Hidroxibenzoatos/farmacología , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Neoplasias Hepáticas/tratamiento farmacológico , Muerte Celular , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Humanos , Lonicera/metabolismo , Sistema de Señalización de MAP Quinasas , Modelos Químicos , Fosforilación , Extractos Vegetales/farmacología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
It is generally believed that the popular nutraceutical 'Kwei Ling Ko' (KLK; Tortoise shell-Rhizome jelly) has antiinflammatory effects, but the mechanism by which its effects are manifested remains unknown. Peroxisome proliferation-activated receptors (PPARs) are members of the nuclear hormone receptor/transcription factor superfamily with multiple roles in adipocyte differentiation, glucose homeostasis, immunomodulation and antiinflammatory regulation. As PPAR is required for adipocyte induction, we used adipogenesis as a possible screen for the activation of the PPAR pathway. Interestingly, an aqueous extract of KLK (sKLK) was able to induce the adipocyte differentiation of fibroblast cell lines. Adipogenesis was confirmed by flow cytometric analysis using a fluorescent lipid stain. Up-regulation of PPARgamma transcripts during adipogenesis was also demonstrated by reverse transcription-polymerase chain reaction (RT-PCR). The sKLK-induced adipogenesis was similar to that elicited by insulin. The activity of nuclear factor-kappaB (NFkappaB), a transcription factor responsible for the regulation of proinflammatory genes, was also down-regulated in response to sKLK. Luciferase reporter gene assays further demonstrated that sKLK inhibited both basal and tumor necrosis factor-alpha-stimulated NFkappaB activation. The activities reported in this study support an immunomodulatory effect for sKLK. As activation of PPAR pathway has a dual role in adipogenesis and anti-inflammation, our observations are consistent with the notion that KLK possesses antiinflammatory properties.
Asunto(s)
Adipogénesis/efectos de los fármacos , Antiinflamatorios/farmacología , Medicamentos Herbarios Chinos/farmacología , Receptores Activados del Proliferador del Peroxisoma/efectos de los fármacos , Smilax , Células 3T3 , Animales , Células Cultivadas , Regulación hacia Abajo , Genes Reporteros/efectos de los fármacos , Humanos , Ratones , FN-kappa B/efectos de los fármacos , Rizoma , Factor de Necrosis Tumoral alfa/farmacología , Tortugas , Regulación hacia ArribaRESUMEN
The bacterial formyl peptide N-formylmethionine-leucine-phenylalanine (fMLP) is a potent chemoattractant for mammalian neutrophils. In this study, we demonstrated the binding of fluorescent dye-conjugated-fMLP to haemocytes of the penaeid shrimp Penaeus penicillatus (Alcock), through the use of flow cytometry. Fluorescence microscopy with rhodamine-fMLP suggested that fMLP receptors are present only in sub-populations of the haemocytes: granulocytes and the semi-granular cells. In addition, fMLP dose-dependently mediated chemotaxis in sub-populations of haemocytes. Microphysiometry experiments demonstrated rapid extracellular acidification upon addition of fMLP, which is in agreement with the observation in neutrophils. t-BOC, the specific fMLP receptor antagonist, was able to block the binding, chemotaxis and extracellular acidification induced by the peptide. The ability of shrimp haemocytes to migrate toward fMLP in vitro suggests that this mechanism may be important for the accumulation of these cells in infected tissues of the shrimps.