RESUMEN
Aleuritolic acid (AA) is a triterpene that is isolated from the root of Croton crassifolius Geisel. In the present study, the cytotoxic effects of AA on hepatocellular carcinoma cells were evaluated. AA exerted dose- and time-dependent cytotoxicity by inducing mitochondria-dependent apoptosis in the hepatocellular carcinoma cell line, HepG2. Meanwhile, treatment with AA also caused dysregulation of autophagy, as evidenced by enhanced conversion of LC3-I to LC3-II, p62 accumulation, and co-localization of GFP and mCherry-tagged LC3 puncta. Notably, blockage of autophagosome formation by ATG5 knockdown or inhibitors of phosphatidylinositol 3-kinase (3-MA or Ly294002), significantly reversed AA-mediated cytotoxicity. These data indicated that AA retarded the clearance of autophagic cargos, resulting in the production of cytotoxic factors and led to apoptosis in hepatocellular carcinoma cells.
Asunto(s)
Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Autofagia/efectos de los fármacos , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Ácidos Palmíticos/farmacología , Células Hep G2 , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Mitocondrias/metabolismoRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Schefflera octophylla (Lour.) Harms, a traditional Chinese herb mainly distributed in Southeast Asia, is extensively prescribed to alleviate pain and treat rheumatoid arthritis (RA), influenza, throat swelling, pain, etc. In this paper, the antinociceptive and anti-inflammatory activities of the ethanol extract and its five different polar fractions of this plant were evaluated. Furthermore, the anti-rheumatoid arthritis activity of the ethanol extract and its active fraction (CHCl3 fraction) were evaluated. And the chemical constituents of the CHCl3 active fraction displayed significant antinociceptive and anti-inflammatory activities were investigated. MATERIALS AND METHODS: Antinociceptive and anti-inflammatory activities were investigated by hot plate test, acetic acid-induced abdominal writhing test and formalin test, xylene-induced ear edema test. The anti-rheumatoid arthritis activity was evaluated through the model of adjuvant-induced arthritis (AA) in rats, paw swelling, pain response, arthritis index and histopathological changes of ankle, the levels of TNF-α, IL-1ß, IL-6 and rheumatoid factor (RF) of rats were detected. The chemical constituents of the CHCl3 fraction were isolated using chromatographic techniques. Their structures were elucidated by spectroscopic data analysis. RESULTS: The results showed that the ethanol extract of S. octophylla has significant dose-dependent anti-inflammatory and antinociceptive activities. And its five different polar fractions especially the CHCl3 fraction significantly inhibited the abdominal writhing induced by acetic acid and ear edema induced by xylene, also increased pain threshold in hot plate test in 120 min and reduced ticking times in formalin test. The ethanol extract of S. octophylla and the CHCl3 fraction demonstrated an anti-RA effect in a dose-dependent manner. The levels of TNF-α, IL-1ß and IL-6 in ethanol extract (600 mg/kg) and CHCl3 fraction (300 mg/kg) groups were significantly lower than those of the model group. The chemical constituents study of the CHCl3 fraction from S. octophylla led to six triterpenoids which were identified as taraxerone (1), 3-epi-taraxerol (2), aleuritolic acid (3), 3-oxofriedelan-28-oic acid (4), 3ß,19α -dihydroxy-urs-12-ene- 24,28-dioic acid (5) and asiatic acid (6). Compounds 1-5 were obtained from this plant for the first time. CONCLUSION: This study proved the antinociceptive, anti-inflammatory and anti-rheumatoid arthritis activities of S. octophylla. Triterpenoids obtained from its CHCl3 fraction may be responsible for those activities. These results could support the fact that S. octophylla is used traditionally to cure inflammatory and pain diseases.
Asunto(s)
Analgésicos/uso terapéutico , Antiinflamatorios/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Edema/tratamiento farmacológico , Dolor/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Ácido Acético , Analgésicos/aislamiento & purificación , Analgésicos/farmacología , Animales , Articulación del Tobillo/patología , Antiinflamatorios/aislamiento & purificación , Antiinflamatorios/farmacología , Araliaceae , Artritis Experimental/sangre , Artritis Experimental/patología , Femenino , Formaldehído , Calor , Interleucina-1beta/sangre , Interleucina-6/sangre , Masculino , Ratones , Dolor/etiología , Fitoterapia , Corteza de la Planta , Extractos Vegetales/química , Extractos Vegetales/farmacología , Raíces de Plantas , Ratas Sprague-Dawley , Factor Reumatoide/sangre , Triterpenos/aislamiento & purificación , Triterpenos/farmacología , Triterpenos/uso terapéutico , Factor de Necrosis Tumoral alfa/sangre , XilenosRESUMEN
The 19-nor-clerodane trans-crotonin (CTN) and the triterpene acetyl aleuritolic acid (AAA), isolated from the stem bark of Croton cajucara Benth (Euphorbiaceae), a traditional medicinal plant from Amazon region of Brazil, as well as the aqueous extract (AE) from its stem bark, were submitted to pharmacological screening for anti-inflammatory and antinociceptive activities in animal models. The oral administration of AAA (50 mg/kg), CTN (50 mg/kg) or AE (300 mg/kg) inhibited the acetic acid-induced writhing in mice. The AE, CTN and AAA had shown significant inhibition of carrageenin-induced edema in rats, in all time intervals measured after the injection of the stimulus, with the greatest inhibition at the first hour for AAA (47.7 percent) and the second hour for CTN (54.4 percent). They have also exhibited significant inhibition in the dextran-induced edema 90 minutes after the stimulus: 31.9 percent for CTN and 28.5 percent for AAA. In the histamine-induced edema, the inhibition showed by CTN and AAA were 43.2 percent and 40.5 percent, respectively, 90 minutes after the injection of stimulus. This study extends and supports the popular medicine and folkloric uses of Croton cajucara in the Amazon region of Brazil.
O 19-nor-clerodano trans-crotonina (CTN) e o triterpeno ácido acetil aleuritólico (AAA), isolados das cascas do caule de Croton cajucara Benth (Euphorbiaceae), uma planta tradicional da Região Amazônica do Brasil, bem como o extrato aquoso (EA) das cascas do caule deste Croton, foram submetidos a experimentos farmacológicos utilizando animais, para avaliação das atividades anti-inflamatória e antinociceptiva. A administração oral de AAA (50 mg/kg), CTN (50 mg/kg) ou AE (300 mg/kg) inibiram as contorções em ratos, induzidas por ácido acético. Os terpenóides AAA e CTN, bem como o extrato polar EA, inibiram significativamente o edema de pata em ratos, induzido por carragenina. As inibições foram observadas em todos os intervalos de medições, tendo sido evidenciado melhores inibições para os terpenóides AAA (47,7 por cento, após a primeira hora) e CTN (54,4 por cento, após a segunda hora). Evidenciou-se ainda, após 90 minutos do estímulo, significante inibição no edema induzido por dextrana (31,9 por cento para CTN e 28,5 por cento para AAA) e por histamina (43,2 por cento para CTN e 40,5 por cento para AAA). Estes resultados confirmam o uso popular de Croton cajucara na região Amazônica do Brasil, no combate a inflamações.
Asunto(s)
Animales , Ratones , Ratas , Antiinflamatorios , Analgésicos/farmacología , Croton , Croton/química , EuphorbiaceaeRESUMEN
A new compound and twelve known compounds were isolated from the ethyl acetate extract of the roots of Homonoia riparia Lour, which are used in folk medicine for treatment of hepatitis, bellyache and scald, by the method of silica gel column chromatography repeatedly with a gradient of PE-EtOAc, PE-Me2CO, CHCl3-Me2CO, CHCl3-MeOH. Their structures were identified as a new compound 1-oxo-aleuritolic acid (1), and twelve known compounds aleuritolic acid (2), 3-acetoxy-aleuritolic acid (3), taraxerone (4), taraxerol (5), methyl 3-acetoxy-12-oleanen-28-oate (6), 3-acetoxy-12-oleanen-28-ol (7), ursolic acid (8), lupenol (9), 3β-acetoxy-lupenol (10), cleomiscosin A (11), chrysophanol (12), and gallic acid (13), which were obtained from this plant for the first time, by the spectroscopic techniques of NMR, HMBC, IR and MS, separately. Among the cytotoxicities evaluation of compounds 1-3 towards AGZY 83-a (human lung cancer cells) and SMMC-7721 (human liver cancer cells) tumor cells was assayed by MTT methods with cis-dichlorodiamminoplatinum (DDP) used as positive control. Compound 2 exerted weak activity against AGZY 83-a with the IC50 value of 33.055 μg·mL-1, while 1 and 3 showed no activity to these two cell lines.