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Cytoprotection by Achyrocline satureioides (Lam) D.C. and some of its main flavonoids against oxidative stress.

Epidemiological studies indicate that dietary antioxidants can influence the incidence of neurodegenerative diseases. Among them flavonoids have been proposed to be effective cytoprotectors. Consequently, herbs with a high concentration of these compounds such as Achyrocline satureioides, Ginkgo biloba and Epilobium parviflorum are of special interest. In this context a comparative study of the cytoprotective capacity of infusions from the three plants against an oxidative insult was performed. Hence, the cytoprotective activity of each infusion against H2O2 injury to PC12 cells was tested and the antioxidant capacity was assessed by the ABTS*+ radical bleaching assay. Free and glycosylated flavonoids contained in the infusions were identified by HPLC and the cytoprotective effect of some of these individual flavonoids was tested. The analysis of the flavonoid content of the infusions revealed different profiles. Epilobium parviflorum infusion showed the highest antioxidant capacity but only Achyrocline satureioides infusion proved to be cytoprotective. Moreover, the free flavonoids quercetin and luteolin contained in this infusion were also cytoprotective. In conclusion, the free radical scavenger capacity did not correlate with the cytoprotective profile of the infusions. The special mixture of unglycosylated Achyrocline satureioides flavonoids could be a clue to explain the unique effect of this plant.

Achyrocline↗

Three different pathways for human LDL oxidation are inhibited in vitro by water extracts of the medicinal herb Achyrocline satureoides.

In this study we investigated the antioxidant properties of one herbal preparation widely used in complementary and alternative medicine in large areas of the world: Achyrocline satureoides (AS), popularly known as "marcela". Although rich in flavonoids, the ethnopharmacological uses of this plant do not include atherosclerosis prevention. Furthermore, no study had been conducted so far exploring the antioxidant activity of Achyrocline satureoides vis-à-vis human LDL oxidation, which is the compelling issue in pinpointing potential cardioprotective new uses for a traditional remedy. We explored the effects of AS extracts on human LDL oxidation, employing 3 different systems which are thought to play a role in oxidation of LDL in the arterial wall: copper, peroxynitrite, and lipoxygenase. Oxidation was monitored by conjugate dienes, TBARS formation and aggregation of apoB using SDS-PAGE. In copper-initiated oxidation a dose dependent inhibition of the initiation and propagation of lipid oxidation is shown by an increase in the lag phase for conjugate diene production which was 60 +/- 15 min in the absence and 120 +/- 20 min in the presence of 4 microg/ml AS extracts (p < 0.001). TBARS production was reduced by 95% after 3 h incubation at 5 microg/ml. Aggregation of apoB was abolished at the same concentrations. SIN-1 (3-morpholinosydnonimine) produces peroxynitrite via generation of NO and O2-. When LDL was incubated in its presence, a milder oxidation was observed as compared with Cu2+, and AS produced over 70% inhibition. Finally, we show a striking dose-dependent inhibitory effect of lipoxygenase conjugate diene production, which is over 95% at AS concentrations of 5 microg/ml. When compared with other antioxidants, AS effect is greater but in the same order of magnitude than that of ascorbic acid and similar to the popular herbal tea Ilex paraguariensis. In all three systems employed an effect is already substantiated at a concentration of the AS extract of 4 microg/ml, which corresponds to a 1/100 dilution of the preparations usually drunk.

Complementary Therapies↗

Evaluation of the antiherpetic activity of standardized extracts of Achyrocline satureioides.

Traditionally, Achyrocline satureioides or 'marcela' has been used in South America for the treatment of several disorders. For the present study, three spray-dried extracts (N1, N2 and N3) were used, all of them prepared with 50% of an hydroethanolic extract rich in flavonoid compounds and 50% of blends of different adjuvants. The cytotoxic concentration which causes destruction in 50% monolayer cells (CC50) was 62.5 microg/ml for the three extracts. The antiviral activity was evaluated by using two different strains of herpes simplex virus (HSV-1) and the best results were obtained with KOS strain and N2 extract. Studies concerning the mechanism of the antiherpetic activity demonstrated that N2 extracts showed no virucidal effect or activity on cellular receptors. HSV-1 DNA synthesis was not inhibited. The antiherpetic activity occurred between the second and ninth hour of the virus replication cycle, probably indicating a perturbation on late stages of this cycle.

Achyrocline↗

Photoprotection by topical application of Achyrocline satureioides ('Marcela').

It has been demonstrated that the use of topical antioxidants can contribute to controling the free radicals excess produced by ultraviolet (UV) irradiation of the skin, being beneficial for processes such as photo aging and eventually cancer. Some plant extracts, particularly because of their polyphenolic constituents, can be beneficial for skin photo damage. Plant extracts of Achyrocline satureioides (AS) and Epilobium parviflorum (EP), potent antioxidant medicinal plants, were mixed with a cosmetic base and applied to the back skin of rabbits. Afterwards the skin was exposed to 1 h of UV irradiation from a known source. The production of the hydroxyl (OH.) radical was assessed in the skin after UV by measuring 2,3-dyhydroxybenzoic acid (2,3-DHBA), produced by the hydroxylation of sodium salicylate, previously injected intracutaneously (i.c.) in the irradiated areas. The UV provoked a marked increase in 2,3-DHBA that was significantly decreased by the AS cosmetic preparation. The EP extract did not show any effect on 2,3-DHBA production. It is concluded that the cosmetic preparation containing the AS extract is able to scavenge OH. production likely to be due to the presence of high concentrations of flavonoid aglycones such as quercetin, luteolin and 3-O-methylquercetin.

Achyrocline↗

Toxicological studies of the aqueous extract from Achyrocline satureioides (Lam.) DC (Marcela).

Achyrocline satureioides (Lam.) DC (Marcela) is known to possess a broad spectrum of pharmacological, medicinal and therapeutic properties. Previous studies have demonstrated various protective abilities of the marcela extracts against various pathological conditions. However, no extensive safety studies have been conducted on these extracts to date. In this paper, we evaluated the acute toxicity (dose levels of 30-300 mg/kg) of an aqueous extract of marcela, administered intraperitoneally and orally in mice and rats. The acute oral maximun tolerable dose in repeated administration during 4 h (1, 3 until 5 g/kg) was also studied in rats. The extract had low acute toxicity when administered intraperitoneally and no toxicity upon oral administration. The LD(50) of aqueous extracts of marcela was found to be greater than 5 g/kg when administered once via gastric intubation to rats. Weight gain, toxicity signs, enzymatic studies (transaminases and phosphatases) and histological evaluation of several organs indicated that the extract was devoid of acute toxicity. These acute studies demonstrated that an aqueous extract of marcela obtained after a 2% infusion is safe and did not cause any detrimental effects in vivo under the conditions investigated in this study.

Achyrocline↗

Antioxidant, a pro-oxidant and cytotoxic effects of Achyrocline satureioides extracts.

In this study we compared the antioxidant properties of five different extracts of different composition obtained from Achyrocline satureioides' inflorescences (Compositae), a widely used Brazilian folk medicinal herb. All of the extracts presented significant antioxidant potential identified by TRAP assay, which increased in the presence of human plasma. Characterization of the content of flavonoids in each extract showed that the FDP80 (ethanol 80%) and FFr (enriched flavonoid fraction) extracts contained a higher content of flavonoids. Cytotoxicity of the extracts as determined in Sertoli cell culture showed that FDP80 and FFr were highly toxic at most concentrations tested. The extracts induced a significant increase in lipid peroxidation levels in Sertoli cells. These results suggest that medicinal herb extracts that contain higher flavonoid concentrations and shows higher antioxidant protection in vitro might not always produce the greatest benefit.

Achyrocline↗

The botanical extracts of Achyrocline satureoides and Ilex paraguariensis prevent methylglyoxal-induced inhibition of plasminogen and antithrombin III.

Endogenously produced dicarbonyls, such as methylglyoxal (MG), are involved in advanced glycation end-product formation and thus linked to the pathophysiology of diabetic chronic complications. While the search for synthetic new antiglycation agents continues, little attention has been paid to putative antiglycation agents in natural compounds. Given the link between glycation and oxidation, in this work, we study the effects of methylglyoxal on two model systems; plasminogen and antithrombin III (AT III), then we set out to unravel a possible antiglycation effect for extracts of the flavonoid-rich common herbal species Achyrocline satureoides (AS) and Ilex paraguariensis (IP). Using SAR-PRO-ARG-pNA as a specific thrombin substrate, we show that incubation of plasma with MG decreases heparin activation of AT III by up to a 70%, in a dose-dependent manner. A parallel dose-dependent decrease in plasminogen activity reaching more than 50% was shown using D-BUT-CHT-lys-pNA as a plasmin-specific substrate. Extracts of AS and IP display a dose dependent inhibition of the action of the dicarbonyl, already significant at a 1/100 dilution of the herbal infusions. The inhibition was comparable to that obtained by using millimolar concentrations of known AGE inhibitors such as aminoguanidine and carnosine as well as micromolar concentrations of the antioxidant ascorbic acid. We believe our system of whole plasma glycation over 16 h with micromolar concentrations of MG, coupled with the measurement of activities of plasminogen and AT III by specific substrates provides a straightforward, practical method for monitoring the action of putative antiglycation agents. If predictably milder glycated forms of AT III and plasminogen were to be secreted in vivo, the loss of activities shown here could act synergistically to generate hyperthrombicity.

Achyrocline↗

Cyclobutane dimers from the Colombian medicinal plant Achyrocline bogotensis.

Five cyclobutane dimers, achyrodimers A-E (1-5), along with 11 known compounds were isolated from the methanol extract of the Colombian medicinal plant Achyrocline bogotensis. Their structures were elucidated by spectroscopy. Several of these compounds were screened for cytokine-inducing activity in human peripheral blood mononuclear cells.

Achyrocline↗

Hepatoprotective activity of Achyrocline satureioides(Lam) D. C.

Aerial parts of Achyrocline satureioides(Lam) D. C. (Asteraceae) are used in folk medicine as infusions or decoctions for the management of several diseases including gastrointestinal and hepatic disorders. These data and the presence of flavonoids and caffeoyl derivatives have led us to study its hepatoprotective and choleretic activities. The hepatoprotective activity was evaluated in the bromobenzene- (BB-) induced hepatotoxicity model in mice through the measurement of the serum levels of alanine-aminotransferase (ALT) and aspartate transaminase (AST), thiobarbituric acid reacting substances (TBARS) and glutathione levels. The aqueous extract of the aerial parts of A. satureioides administered before BB, at the dose of 300 mg kg(-1)p.o., demonstrated significant inhibition (P< 0.01) in the BB increase of liver ALT and AST and in the BB-induced increase of liver TBARS content. Also it was able to significantly increase (P< 0.05) the depleted levels of liver glutathione. In addition, at the same dose, a significant increase (P< 0.01) in the bile flow of rats was found. The results obtained with the aqueous extract of A. satureioides support its use in popular medicine as a hepatoprotective and digestive agent, and the effects might be mediated through the antioxidant and choleretic activities.

Alanine Transaminase↗

Immunologically active metallic ion-containing polysaccharides of Achyrocline satureioides.

Two homogeneous, metallic ion-containing pectic polysaccharides with mean M(r)s of 7600 and 15,000 were isolated from dried aerial parts of Achyrocline satureioides by anion exchange column chromatography on DEAE-Sepharose CL-6B and gel filtration column chromatography on Fractogel TSK HW-50 (S). The structures, as determined by methylation analysis, carboxyl reduction, and partial acid hydrolysis, were shown to be rhamnogalacturonans. Both pectins show a pronounced anticomplementary effect in vitro. The larger carbohydrate AS 4 of higher M(r) exerts anti-inflammatory activity and a strong enhancement of phagocytosis in vivo.

Carbohydrate Sequence↗

Mutagenic and genotoxic effects of aqueous extracts of Achyrocline satureoides in prokaryotic organisms.

Aqueous extracts of Achyrocline satureoides (Marcela and/or Macela) were tested for the presence of genotoxic activity in microorganisms. This species belongs to the family Compositae and is used on a large scale by the population of South Brazil. The extracts showed genotoxic activity in the presence of S9 mix in the Ames test TA100, TA98 and TA102 strains, 'SOS' spot chromotest and Microscreen phage-induction assay. The positive results were related to the presence of quercetin and caffeic acid in the aqueous extracts.

Animals↗

Pharmacological investigations on Achyrocline satureioides (LAM.) DC., Compositae.

Achyrocline satureioides (Lam.) DC. inflorescences have been used as remedies in folk medicine for the treatment of a variety of human ailments, particularly those of the gastrointestinal tract. Different extracts of inflorescences have been tested for anti-inflammatory, analgesic, antispasmodic, constipating and sedative activities. The aqueous extracts (maceration and decoction) and ethanolic macerate exhibited an inhibition of the carrageenan-induced rat paw oedema at a dose range of 75-500 mg kg-1 i.p., and also showed analgesic effect with the acetic acid-induced writhing test in mice. The gastrointestinal propulsion of a charcoal suspension was not affected significantly by any extract, at a dose of 200 mg kg-1 p.o., in mice. The aqueous decoction increased pentobarbital-induced sleeping time, at doses of 200 and 500 mg kg-1 i.p. and p.o., in mice. The ethanolic macerate inhibited contractions induced by acetylcholine, histamine, noradrenaline and barium chloride in four different smooth muscle tissues. The antispasmodic and anti-inflammatory activities were reproduced with quercetin, luteolin and quercetin 3-methyl ether, flavonoids that have been isolated from this plant. A partial evaluation of the toxicity of the extracts was also performed. The pharmacological effects assayed are discussed in relation to the chemical constituents of this plant and its popular use in gastrointestinal disturbances, and inflammatory conditions could be related to the presence of the flavonoids.

Analgesics↗

Effects of hydroalcoholic extracts of Portulaca pilosa and Achyrocline satureioides on urinary sodium and potassium excretion.

Porulaca pilosa has been used in Brazil as a traditional remedy to cause diuresis, antipyresis and analgesia. Achyrocline satureioides has been used in folk medicine as antiinflammatory, hypoglycemic, sedative and to treat gastrointestinal disorders such as diarrhea and dysentery suggesting that it may affect salt and water reabsorption by the gastrointestinal tract. In the current study, hydroalcoholic extracts of both plants were investigated in order to examine their renal effects. The results support the claim that extracts of P. pilosa present renal effects but not the popular belief that it affects diuresis. It has also been provided that, in rats, it causes an increase in K excretion without a concomitant change in water diuresis or Na excretion. Our findings also support the popular belief that A. satureioides does not apparently have renal effects and it might change renal ion transport based on observations that it affects gastrointestinal reabsorption.

Animals↗

LC determination of flavonoids: separation of quercetin, luteolin and 3-O-methylquercetin in Achyrocline satureioides preparations.

The pharmacological activities of the flavonoids show the interest in quantifying these constituents in phytopharmaceutical preparations, as well as in the validation of the analytical methodologies. LC methods have been reported to quantify isolated flavonoids or these compounds in complex biological matrices, such as herbal raw materials and extractive preparations. This work was designed, therefore, to develop an LC system to separate quercetin, luteolin and 3-O-methylquercetin and to quantify them in extractive solutions from Achyrocline satureioides. The main validation parameters of the method are also determined. The method showed linearity for quercetin and luteolin in the range 1-10 microg/ml. The aqueous and ethanol 80% extractive solutions showed linear response in the range 2.5-20 microl/ml and ethanol 40% extractive solution in the range 2.5-10 microl/ml. Precision and accuracy were determined for ethanol 80% extractive solution, in the concentration of 10 microl/ml. The LC method showed an excellent performance in separating the flavonoids quercetin, luteolin and 3-O-methylquercetin in A. satureioides extracts, since the presence of interference has been previously evaluated.

Calibration↗

Achyrocline satureioides essential oils from southern Brazil and Uruguay.

The essential oils of the aerial parts of Achyrocline satureioides (D.C.) Lam., a regional medicinal plant, from different collection locations in the South of Brazil and Uruguay were examined by GC and GC-MS. Monoterpene and sesquiterpene hydrocarbons were the main fractions of both the oils from Brazil and Uruguay. The oxygenated monoterpenes and sequiterpenes were at much lower percentage in both samples. The Uruguayan samples have 1,8-cineole which is not present in the Brazilian samples and has not been reported in other samples from Brazil. The results indicated a high biodiversity of the native populations of A. satureioides.

Asteraceae↗