Antiangiogenic Activity of n-hexane Insoluble Fraction and Its Tylophorine Component from Ficus septica Leaves in Chicken Chorioallantoic Membrane Induced by bFGF.

OBJECTIVE: Ficus septica is an Indonesian medicinal plant traditionally used to treat various illness, including cancer. The n-hexane insoluble fraction of the ethanolic extract of F. septica leaves (HIFFS) shows a potential anticancer activity against breast cancer cell line T47D. Considering that angiogenesis is a pivotal factor in malignant cancer growth, progression, and invasion, we aimed to investigate the antiangiogenic effect of HIFFS on chicken chorioallantoic membrane (CAM) induced by bFGF. We also evaluated tylophorine, the cytotoxic alkaloid of F. septica. METHODS: Chicken CAM was used to assess the antiangiogenic effect. Fertilized chicken eggs were induced with basic fibroblast growth factor (bFGF) ex ovo. Prior to bFGF induction, HIFFS (2.33, 4.65, 6.98, and 9.30 µg/mL) or tylophorine (9.20 µM) was added (10 µL) to a paper disk and implanted to the CAM. After 48 h of incubation, each treatment group was photographed, and the number of new blood vessel was calculated and compared with that in the solvent-treated group to determine the antiangiogenic activity. Histology of the CAM was evaluated after hematoxylin-eosin and Mallory acid fuchsin staining. RESULTS: We found that HIFFS at low concentrations (2.33, 4.65, 6.98, and 9.30 µg/mL) inhibited angiogenesis activity (31.87, 41.99, 53.65, and 70.08, respectively) in chicken CAM induced by bFGF. Tylophorine (9.20 µM) also showed similar antiangiogenesis activity in the same model. Histopathology analysis revealed that HIFFS and tylophorine reduced the number of new blood vessels in CAM induced by bFGF. CONCLUSION: HIFFS and tylophorine showed antiangiogenic effect on chicken CAM induced by bFGF. This finding emphasized the potential of F. septica as a candidate anticancer agent.

Pharmacokinetic Herb-Drug Interactions of Glipizide with Andrographis paniculata (Burm. f.) and Andrographolide in Normal and Diabetic Rats by Validated HPLC Method.

Co-administered medicinal herbs can modify a drug's pharmacokinetics (PK), effectiveness, and toxicity. Andrographis paniculata (Burm. f.) ethanolic extract (APE) and andrographolide (AND) (a potent CYP2C9 inducer/inhibitor) can alter the pharmacokinetic parameters of glipizide (GLZ). This study aimed to determine the potential pharmacokinetics of herb−drug interactions between GLZ and APE/AND in the plasma of normal and diabetic rats using the HPLC bioanalysis method. The glipizide bioanalytical method established with RP-HPLC/UV instrument was validated following the EMA guidelines. GLZ was administered alone and in combination with APE or AND to normal and diabetic rats. The GLZ pharmacokinetic parameters were estimated according to the correlation between concentration and sampling time using the PK solver program. A simple and rapid GLZ bioanalysis technique with a lower limit of quantitation of 25 ng/mL was developed and presented the following parameters: accuracy (error ≤ 15%), precision (CV ≤ 15%), selectivity, stability, and linearity (R2 = 0.998) at concentrations ranging 25−1500 ng/mL. APE administration significantly improved the Cmax and AUC0−t/AUC0−∞ GLZ values in normal and diabetic rats (p < 0.05). AND significantly reduced the bioavailability of GLZ in diabetic rats with small values of T 1/2, Cmax, and AUC0−t/AUC0−∞ (p < 0.05). This combination can be considered in administering medications because it can influence the pharmacological effects of GLZ.

Carica papaya leaf extract inhibits prostatitis-associated prostatic hyperplasia via the TRAF6/TAK1/MEK/NF-κB pathway.

Prostatitis, defined as a pathological inflammatory change in the prostate tissue, is one of the most prevalent urological conditions in men. However, optimal management of prostatitis remains unclear, and treatment outcomes are unsatisfactory owing to adverse effects. Carica papaya leaf extract (PAL) is known for its antioxidant, immunomodulatory, and anticancer properties; however, evidence of its anti-inflammatory effect in prostatic tissues remains elusive. In this study, the therapeutic effects and underlying molecular mechanisms of PAL in mice with experimental autoimmune prostatitis (EAP) and a prostatic cell line (RWPE-1 cells) exposed to inflammatory conditioned medium were investigated. PAL suppressed pathological alterations in EAP and markedly reduced prostate weight in EAP mice. Histological analysis revealed that PAL alleviates prostatic hyperplasia. Furthermore, PAL significantly reduced cyclooxygenase-2 mRNA and protein expression; production of inflammatory cytokines, including interleukin-6, tumor necrosis factor-α, and transforming growth factor-ß; and TRAF6/TAK1/MEK/ERK and NF-κB pathway-related protein expression. TRAF6/TAK1/MEK/ERK and NF-κB pathway-related proteins were upregulated in inflammatory conditioned medium-stimulated RWPE-1 cells, but PAL reduced the expression of these markers. Particularly, PAL treatment suppressed the nuclear translocation of NF-κB p65 and phosphorylation of p65 in RWPE-1 cells exposed to the inflammatory conditioned medium. Collectively, the results demonstrate the anti-proliferative and anti-inflammatory effects of PAL in the experimental prostatitis model, which highlights the potential of PAL as a new therapeutic agent in the treatment of prostatic disease.

Ginger extract and its compound, 6-shogaol, attenuates painful diabetic neuropathy in mice via reducing TRPV1 and NMDAR2B expressions in the spinal cord.

ETHNOPHARMACOLOGICAL RELEVANCE: In silico data revealed that the active compound of ginger (Zingiber officinale Roscoe), 6-shogaol, has strong affinity toward transient receptor potential vanilloid-1 (TRPV-1). TRPV-1 is expressed in nervous tissue and pancreatic ß-cells. Prolonged induction of TRPV-1 is related to the expression of N-methyl-D-aspartate receptor subunit 2B (NMDAR2B). However, there are no data on TRPV-1 and NMDAR2B expressions in nervous tissue after 6-shogaol or ginger extract treatment nor pancreatic islet morphology and insulin expression in mice model of painful diabetic neuropathy (PDN). AIM OF THE STUDY: This study aimed to investigate the mechanism of action of ginger extract and its compound, 6-shogaol, on pancreatic islets as well as on expressions of TRPV-1 and NMDAR2B in the spinal cord of streptozotocin (STZ)-induced mice model of PDN. MATERIALS AND METHODS: Sixty-four 5-6 weeks old male-Balb/C mice were induced with 110 mg/kgBW STZ i.p., while eight mice were used as control group. Mice with blood glucose level ≥200 mg/d, that suffered hyperalgesia and allodynia were classified as PDN mice. Hot plate and von Frey filament tests were performed once a week until termination. At day 28 after considered as PDN, ginger extracts, 6-shogaol or gabapentin as control treatment were given once daily for 21 days until day 49, except for the diabetic control group. Upon termination, mice' pancreas were fixed, processed as paraffin sections and stained with hematoxylin eosin. Total volume of pancreatic islets was estimated using Cavalieri methods. Immunohistochemistry on pancreatic sections were performed to observe insulin expression. mRNA was extracted from lumbar segments of the spinal cord, followed by cDNA preparation and quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) to measure the expressions of TRPV1 and NMDAR2B. The mean differences between groups were analyzed using one-way analysis of variance (ANOVA) with p < 0.05 considered statistically significant. RESULTS: Ginger extracts and 6-shogaol alleviated hyperalgesia and allodynia. The groups that received ginger extract 400 mg/kgBW or 6-shogaol 15 mg/kgBW had significantly lower TRPV1 and NMDAR2B expressions in the spinal cord compared to the diabetic control group (p < 0.001; p < 0.05). However, no differences in volume of pancreatic islets (p > 0.05) nor insulin expression were observed in all PDN groups. CONCLUSION: Ginger extracts and its compound, 6-shogaol, reduced pain symptoms in PDN via its effect on decreasing TRPV1 and NMDAR2B expressions in the spinal cord, with very limited effect on pancreatic islets.

Corrigendum to "Antiadipogenic Effects of Aster glehni Extract: In Vivo and In Vitro Effects".

[This corrects the article DOI: 10.1155/2013/859624.].

Anti-inflammatory effects of an ethanol extract of Aster glehni via inhibition of NF-κB activation in mice with DSS-induced colitis.

Although Aster glehni has been reported to prevent diabetes mellitus, hypercholesterolemia, insomnia, and cardiovascular disease, the anti-inflammatory effect of Aster glehni in colonic tissue remains unclear. In this study, we investigated the anti-inflammatory effects and the underlying molecular mechanism of an ethanol extract of Aster glehni (AG) in mice with dextran sulfate sodium (DSS)-induced colitis. AG significantly attenuated DSS-induced DAI scores, which implied that it suppressed diarrhea, gross bleeding, and the infiltration of immune cells. AG administration also effectively prevented shortening of the colon length and enlargement of the spleen size. Histological examinations indicated that AG suppressed colonic damage and the thickness of the muscle layer induced by DSS. In addition, AG inhibited the production of pro-inflammatory cytokines, such as TNF-α, IL-1ß, and IL-6, and the protein expression of COX-2 and iNOS in mice with DSS-induced colitis. Administration with AG suppressed the activation of nuclear factor-κB (NF-κB) including the nuclear translocation of the p65 NF-κB subunit, phosphorylation and degradation of IκB-α. Taken together, these findings suggest that the anti-inflammatory effects of AG are mainly related to the inhibition of the expressions of inflammatory mediators via NF-κB inactivation, and support its possible therapeutic application in colitis.

Hypotensive Activity of Ethanolic Extracts of Morinda citrifolia L. Leaves and Fruit in Dexamethasone-Induced Hypertensive Rat.

The effect of ethanolic extract of Morinda citrifolia leaves and fruit on blood pressure in dexamethasone-induced hypertension rat was evaluated. Total phenolic content of Morinda citrifolia leaves ethanolic extract (MCLEE) and Morinda citrifolia leaves ethanolic extract (MCFEE) was 1.789 ± 0.116 and 1.677 ± 0.051 mg of gallic acid equivalents per gram sample, respectively. Rutin level in MCLEE was 0.92 ± 0.19%, and scopoletin level in MCFEE was 0.46 ± 0.05%. MCLEE, MCFEE, and its extract combination significantly decreased the blood pressure of hypertensive rats. The combination group showed highest hypotensive activity by lowering systolic blood pressure by 16.71 ± 3.95%, diastolic blood pressure by 21.49 ± 7.90%, and mean arterial blood pressure by 19.58% ± 6.35. All extract treatments have not been able to repair or inhibit renal damage caused by dexamethasone induction.

Gastroprotective Effect of Combination of Hot Water Extracts of Licorice (Glycyrrhiza glabra), Pulasari Stem Bark (Alyxia reinwardtii), and Sembung Leaf (Blumea balsamifera) Against Aspirin-Induced Gastric Ulcer Model Rats.

Licorice (Glycyrrhiza glabra), Pulasari stem bark (Alyxia reinwardtii) and Sembung leaf (Blumea balsamifera) are traditionally used to treat gastrointestinal disorders. The aim of the study was to investigate gastroprotective effect of hot water extracts combination of those herbal against aspirin-induced gastric ulcer model in rats. The combination consisted of fixed doses of Licorice 273 mg/kg BW and Sembung leaf 457.5 mg/kg BW, and also consisted of Pulasari stem in various doses i.e. 100 mg/kg BW (first group), 200 mg/kg BW (second and sixth group) and 300 mg/kg BW (third group). The fourth grup rats received sucralfate 360 mg/kg BW. Ten minute after seven consecutive days of drug administration, the rats were induced with aspirin 450 mg/kg BW except sixth group rats. The fifth group rats only received aspirin without any protective agents. The number and area of gastric ulcers were evaluated macroscopically. Whereas, histopatological observation was used for evaluation of mucosal damage score, and the number of eosinophils and mast cells. In the study, herbal extracts combination markedly exhibited protective effects indicated by less number and smaller area of gastric ulcers in comparison to those of aspirin group (P < 0.05). The score of mucosal damages were also decreased in herbal extracts combination groups. The number of eosinophils and mast cells of herbal combination groups were observed to be smaller than those of aspirin group (P < 0.05). In conclusion, herbal combination of Licorice (Glycyrrhiza glabra), Pulasari stem bark (Alyxia reinwardtii) and Sembung leaf (Blumea balsamifera) is potential to develop as a gastroprotective agent.

Isoeugenin, a Novel Nitric Oxide Synthase Inhibitor Isolated from the Rhizomes of Imperata cylindrica.

Phytochemical studies on the constituents of the rhizomes of Imperata cylindrica (Gramineae) were performed using high-performance liquid chromatography (HPLC). We also aimed to search for any biologically active substance capable of inhibiting nitric oxide (NO) formation in lipopolysaccharide (LPS)-activated macrophage 264.7 cells, by testing four compounds isolated from this plant. Four compounds, including a new chromone, isoeugenin, along with ferulic acid, p-coumaric acid, and caffeic acid were isolated and identified by NMR spectroscopy. The structure of isoeugenin was determined as 7-hydroxy-5-methoxy-2-methylchromone by the 2D-NMR technique. Among the four compounds, isoeugenin has the lowest IC50 value on the inhibition of NO production in LPS-activated macrophage RAW264.7 cells (IC50, 9.33 µg/mL). In addition, isoeugenin significantly suppressed the LPS-induced expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and proinflammatory cytokines mRNA levels. Taken together, these results suggest that the anti-inflammatory activity of isoeugenin is associated with the down-regulation of iNOS, COX-2, and pro-inflammatory cytokines in RAW264.7 cells. Accordingly, our results suggest that the new chromone isoegenin should be considered a potential treatment for inflammatory disease.

Cytotoxic Effect and Constituent Profile of Alkaloid Fractions from Ethanolic Extract of Ficus septica Burm. f. Leaves on T47D Breast Cancer Cells.

The study aimed to investigate the profile of alkaloids in two ethyl acetate soluble fractions, namely fractions A and B from an ethanolic extract of Ficus septica leaves and cytotoxic effect on T47D breast cancer cells. Preparation of both fractions involved maceration of leaves with 70% (v/v) ethanol, filtration with Al2O3, precipitation with 0.1 N HCl, Mayer reagent, and 0.1 N NaOH, and also partition with ethyl acetate. Qualitative thin layer chromatography (TLC) was conducted to determine the profile of alkaloids in the two fractions, using alkaloid specific reagents such as Dragendorff, sodium nitrite, and Van Urk-Salkowski. Cytotoxic effects of both fractions on T47D cells were evaluated using MTT assay with a concentration series of 1.56; 3.12; 6.25; 12.5; 25 and 50 µg/mL. The TLC test showed that fractions A and B contained alkaloids with Rx values of 0.74 and 0.80 for fraction A and 0.74, 0.84, 0.92 for fraction B with regard to yohimbine using the mobile phase of n-buthanol:glacial acetic acid:distilled water (3:1:1 v/v/v). Moreover, an indole alkaloid was detected with Rx values of 0.80 and 0.84, respectively. Fractions A and B exhibited high cytotoxic effects on T47D cells with IC50 values of 2.57 and 2.73 µg/mL, respectively. In conclusion, overall the results of this study showed that fractions of Ficus septica contain alkaloids including indole alkaloid or its derivatives and possess a cytotoxic effect on T47D cells. This research supports the idea that alkaloids in F. septica have anticancer activity.

Immunomodulatory Effects of Ethanolic Extract of Thyphonium flagelliforme (Lodd) Blume in Rats Induced by Cyclophosphamide.

The present study aimed to examine the immunomodulatory effect of ethanolic extract of Typhonium flagelliforme (Lodd) Blume in cyclophosphamide-treated rats. The immunomodulatory effects were determined by lymphocytes proliferation, phagocytic activity of macrophages, plasma cytokines of tumor necrosis factor-α, interleukin-1α, interleukin-10 levels, and killer T cells (CD8+ T cells) counts. The results showed that the administration of ethanolic extract of T flagelliforme reduced immunosupessive effect on lymphocyte proliferation, increase the number and phagocytic activity of macrophages in cyclophosphamide-treated rats. Moreover, the ethanolic extract of T flagelliforme also significantly (P < .05) improved the immune system activities especially the proliferation of CD8+T cells and reduced the suppressive effects on cytokines such as tumor necrosis factor-α and interleukin-1α. In conclusion, the ethanolic extract of T flagelliforme has immunomodulatory properties in cyclophosphamide-treated rats. The results suggest that T flagelliforme can reduce immunosuppresive effect caused by a chemotherapeutic agent.

Quantification of major compounds from Ixeris dentata, Ixeris dentata Var. albiflora, and Ixeris sonchifolia and their comparative anti-inflammatory activity in lipopolysaccharide-stimulated RAW 264.7 cells.

The aim of the present study was to evaluate the comparative anti-inflammatory activities of Ixeris dentata (ID), Ixeris dentata var. albiflora (IDA), and Ixeris sonchifolia (IS) and to identify the main compounds present in extracts. The anti-inflammatory activity was evaluated through lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 murine macrophages. Five main compounds consisting of chlorogenic acid, caffeic acid, luteolin 7-O-glucoside, luteolin 7-O-glucuronide, and luteolin were used for simultaneous high-performance liquid chromatography quantification. The total phenolic content present in ID (30 mg/g GAE), IDA (35.33 mg/g GAE), and IS (43.79 mg/g GAE) was correlated to the corresponding LPS-induced NO production inhibitory effect in RAW 264.7 cells as expressed with IC(50) values 26.19, 21.43, and 7.59 µg/mL, respectively. Luteolin 7-O-glucoside was found as the major compound in ID (8.76 mg/g dry weight) and IDA (10.35 mg/g dry weight) and luteolin 7-O-glucuronide was the major compound in IS (34.66 mg/g dry weight). Luteolin 7-O-glucoside and luteolin 7-O-glucuronide inhibited LPS-induced NO production with IC(50) values of 30 and 4.5 µM, respectively. Furthermore, luteolin, luteolin 7-O-glucoside, and luteolin 7-O-glucuronide suppressed the expression of iNOS and COX-2, and t-BHP-induced ROS generation in LPS-stimulated RAW 264.7 cells. These results clearly showed that the anti-inflammatory potential of ID, IDA, and IS extract are primarily due to their contents of luteolin 7-O-glucoside and luteolin 7-O-glucuronide, respectively.

Simultaneous quantification and validation of new peroxynitrite scavengers from Artemisia iwayomogi.

CONTEXT: Artemisia iwayomogi Kitamura (Compositae) has been very widely used for the treatment of acute or chronic hepatitis, jaundice, and gastritis. In the course of our continuing efforts to identify and quantify peroxynitrite scavengers from Compositae plants, A. iwayomogi was used in this study. OBJECTIVE: The present study was aimed to identify and quantify the peroxynitrite scavengers of A. iwayomogi. MATERIALS AND METHODS: Silica gel and ODS were used for column chromatography. The isolated compounds were quantified using an HPLC equipped with a Capcell Pak C18 column (5 µm, 250 mm × 4.6 mm i.d.), and the method was validated for the quality control. Peroxynitrite (ONOO(-))-scavenging activities of the compounds and extracts were evaluated on the measurement of highly fluorescent rhodamine 123 converted from non-fluorescent dihydrorhodamine (DHR)-123 under the presence of peroxynitrite. RESULTS: Based on the spectroscopic evidences, a new compound, 2"-O-caffeoylrutin (2"-O-trans-caffeic acid ester of quercetin 3-O-α-L-rhamnopyranosyl(1 → 6)-ß-D-glucopyranoside) was isolated and determined together with patuletin 3-O-glucoside, scopolin, scopoletin, rutin, 3,4-dicaffeoylquinic acid, and chlorogenic acid. All of them were potent peroxynitrite scavengers (IC50 ≤ 1.88 µg/mL). DISCUSSION AND CONCLUSION: The peroxynitrite scavengers were mainly distributed in the EtOAc fraction rather than the ether and BuOH fractions. The 70% MeOH extract exhibited a high peroxynitrite-scavenging activity. Through the validation, the present HPLC method was verified to be sufficiently sensitive, accurate, precise, and stable. Therefore, this method can be used for the quality control of A. iwayomogi.

Simultaneous quantification and peroxynitrite-scavenging activities of flavonoids in Polygonum aviculare L. herb.

The plant Polygonum aviculare L. (Polygonaceae) is an annual herbaceous plant which is known to be beneficial for treating gastroduodenal ulcer, hypertension, diarrhea, hemorrhage, and hemorrhoids. Ten phenolic compounds, including nine flavonoids (myricetin, quercetin, kaempferol, myricitrin, desmanthin-1, isoquercitrin, quercitrin, avicularin, juglanin), and gallic acid were used for simultaneous HPLC quantification and peroxynitrite-scavenging assay. Simultaneous quantification of these substances were performed on five extracts (EtOH-, MeOH-, 70% MeOH-, 30% MeOH-, and H2O extracts) as well as on the three fractions (Et2O-, EtOAc-, and BuOH fractions), under the condition of a Capcell Pak C18 column (5µm, 250mm×4.6mm i.d.) and a gradient elution of 0.05% trifluoroacetic acid (TFA) and MeOHCH3CN (60:40). Of the three fractions, the EtOAc fraction displayed the highest content of flavonoids (sum, 208.9mg/g) with the strongest peroxynitrite-scavenging activity (IC50, 2.68µg/mL). The activities of the eight compounds (myricitrin, isoquercitrin, avicularin, quercitrin, myricetin, desmanthin-1, quercetin, and kaempferol) were comparable to that of the positive control (l-penicillamine; IC50: 1.03µg/mL). These results suggest that folkloric medicinal uses of P. aviculare are mainly attributed to flavonoids, such as particularly highly contained myricetin, myricitrin, and desmanthin-1.

Simultaneous analysis and peroxynitrite-scavenging activity of galloylated flavonoid glycosides and ellagic acid in Euphorbia supina.

The herbs of Euphorbia supina (Euphorbiaceae) have been used to treat hemorrhage, chronic bronchitis, hepatitis, jaundice, diarrhea, gastritis, and hemorrhoids as a medicinal herb. This work is aimed to qualitatively and quantitatively analyze the polyphenols with peroxynitrite-scavenging activities. The eight compounds: gallic acid, methyl gallate, avicularin, astragalin, juglanin, isoquercitrin 6″-gallate, astragalin 6″-gallate, and ellagic acid, were isolated from E. supina and used for HPLC analysis and peroxynitrite (ONOO(-))-scavenging assay. Simultaneous analysis of the eight compounds was performed on MeOH extract and its fractions. The contents in MeOH extract and peroxynitrite-scavenging activities of the dimer of gallic acid, ellagic acid (15.64 mg/g; IC50 0.89 µM), and two galloylated flavonoid glycosides, astragalin 6″-gallate (13.72 mg/g; IC50 1.43 µM) and isoquercitrin 6″-gallate (16.99 mg/g; IC50 1.75 µM), were high, compared to other compounds. The legendary uses of E. supina could be attributed to the high content of polyphenols, particularly ellagic acid, isoquercitrin 6″-gallate, and astragalin 6″-gallate as active principles.

Antiadipogenic Effects of Aster glehni Extract: In Vivo and In Vitro Effects.

Aster glehni (AG) is a Korean traditional herb that grows in Ulleungdo Island, Republic of Korea. None of the several reports on AG include a determination of the effect of AG on adipogenesis. The primary aim of this study was to determine whether AG attenuates adipogenesis in mouse 3T3-L1 cells and epididymal fat tissue. AG blocked the differentiation of 3T3-L1 preadipocytes in a concentration-dependent manner and suppressed the expression of adipogenesis-related genes such as PPAR γ , C/EBP α , and SREBP1c, the master regulators of adipogenesis. Male C57BL/6J mice were divided randomly and equally into 4 diet groups: control diet (CON), high-fat diet (HFD), HFD with 1% AG extract added (AG1), and HFD with 5% AG extract added (AG5). The experimental animals were fed HFD and the 2 combinations for 10 weeks. Mice fed HFD with AG gained less body weight and visceral fat-pad weight than did the mice fed HFD alone. Moreover, AG inhibited the expression of important adipogenic genes such as PPAR γ , C/EBP α , SREBP1c, LXR, and leptin in the epididymal adipose tissue of the mice treated with AG1 and AG5. These findings indicate antiadipogenic and antiobesity effects of AG and suggest its therapeutic potential in obesity and obesity-related diseases.

Cardioprotective and hepatoprotective effects of Citrus hystrix peels extract on rats model.

OBJECTIVE: To observe the combination effect of doxorubicin and Citrus hystrix (kaffir lime's) peel ethanolic extract (ChEE) on blood serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity and cardio-hepato-histopathology of female Sprague Dawley rats. METHODS: Doxorubicin and ChEE (5 rats per group) were administered in five groups of 3 rats each for 11 d. Group I: doxorubicin (dox) 4.67 mg/kg body weight; Group II: dox+ChEE 500 mg/kg body weight; Group III: dox+ChEE 1 000 mg/kg body weight; Group IV: ChEE 1 000 mg/kg body weight; Group V: untreated (control). RESULTS: ChEE repaired cardiohistopathology profile of doxorubicin induced cardiotoxicity and hepatotoxicity rats, but did not repair neither hepatohistopathology profile nor reduce serum activity of ALT and AST. CONCLUSION: ChEE has potency to be developed as cardioprotector agent in chemotherapy.

Simultaneous quantification and validation of caffeoylquinic acids and flavonoids in Hemistepta lyrata and peroxynitrite-scavenging activity.

Traditionally, Hemistepta lyrata is consumed as a mountainous vegetable or a medicinal herb to treat inflammation, fever, hemorrhage, and hemorrhoids. In order to provide the scientific evidence of traditional uses of this plant, we identified and quantified thirteen active substances (caffeic acid, chlorogenic acid, and 3,5-di-O-caffeoylquinic acid as caffeoylquinic acids; apigenin, isorhoifolin, acacetin, linarin, diosmetin, diosmin, pectolinarigenin, and pectolinarin as flavones or their glycosides; kaempferol 3-O-rutinoside and rutin as flavonol glycosides) from H. lyrata and evaluated their peroxynitrite-scavenging activity. The chromatographic separation was performed on a Capcell Pak C18 column (5µm, 250mm×4.6mm i.d.) with a gradient elution of 0.05% TFA (trifluoroacetic acid) and 0.05% TFA in MeOH-CH(3)CN (60:40). Validation of HPLC methods on the linearity, LOD, LOQ, intra-day and inter-day variabilities, recovery, and repeatability proved that this method is selective, sensitive, precise, accurate, and reproducible. In peroxynitrite-scavenging assay, caffeic acid derivatives (chlorogenic acid, caffeic acid, and 3,5-di-O-caffeoylquinic acid) exhibited relatively lower IC(50) values than other substances tested. And HPLC simultaneous quantification showed that the 70% MeOH extract and the BuOH fraction contain a higher quantity of caffeic acid derivatives (17.82 and 30.09mg/g, consecutively). Therefore, caffeic acid derivatives could be the main contributors to the peroxynitrite-scavenging activity of H. lyrata than other phenolic substances.

Anti-diabetic effect of a combination of andrographolide-enriched extract of Andrographis paniculata (Burm f.) Nees and asiaticoside-enriched extract of Centella asiatica L. in high fructose-fat fed rats.

Traditionally, a combination of medicinal plants is commonly used for lowering blood glucose in diabetic patients in order to provide additional benefits of the single drug. A. paniculata and C. asiatica are two traditional medicines form South Asian and Southeast Asain countries consumed by people for treating daibates mellitus and its complications. Hyperglycemia in the rats was stimulated by high fructose-fat diet that consists of 36% fructose, 15% lard, and 5% egg yolks in 0.36 g/200 g body weight for 70 days. The rats were orally administered with the combination of andrographolide-enriched extract of A. paniculata (AEEAP) leaves and asiaticoside-enriched extract of C. asiatica (AEECA) herbs from day 70 for 7 days. Antidiabetic activity was evaluated by estimating mainly the blood glucose levels and other parameters such as HDL, LDL, cholesterol and triglyceride. The results showed that combination at the ratio of 70:30 exhibited a promosing antidiabetic effect in high-fat-fructose-fed rat, and exhibited sinergistic effects on blood cholesterol and HDL levels. It can be concluded that its antidiabetic effect was better than that of single treatment of AEEAP or AEECA. That combination was also potential to develop as a blood glucose-lowering agent for diabetic patients.

Synergistic effects of ethyl acetate fraction of Ficus septica Burm. f. and doxorubicin chemotherapy on T47D human breast cancer cell line.

OBJECTIVE: Previously, ethanolic extract of Ficus septica Burm. f. (Moraceae) leaves and its ethyl acetate soluble fraction (EASF) exhibited potent cytotoxic effects on T47D breast cancer cells. In the present study, we further investigated the effects of EASF of ethanolic extract of F. septica leaves in combination with doxorubicin on T47D breast cancer cell line in cytotoxicity, cell cycle arrest and apoptosis induction. METHODS: The cytotoxic effect analysis on T47D cells was carried out using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Analysis of cell cycle distribution was performed using a flowcytometer and the data were analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethidium bromide-acridin orange. The expression of cleaved-poly ADP-ribose polymerase in the T47D cell line was identified using immunohistochemical techniques. RESULTS: The combination of doxorubicin (2 to 8 nmol/L) with EASF (0.875 to 7 µg/mL) more potently inhibited cell growth than the single treatment of doxorubicin in T47D cells. In addition, the combination of doxorubicin and EASF could increase the incidence of cells undergoing apoptosis. EASF was found to improve cytotoxic effect of doxorubicin by changing the inhibition of cell cycle G(2)/M to G(1) phase. The combination also exhibited a more intensive stimulatory effect on cleaved-PARP expression in T47D cells than the single treatment. CONCLUSION: It is concluded that EASF may enhance doxorubicin activities in T47D cells by inducing apoptosis and cell cycle arrest.