Gallic acid exhibits risks of inducing muscular hemorrhagic liposis and cerebral hemorrhage--its action mechanism and preventive strategy.

Gallic acid (3,4,5-trihydroxybenzoic acid) (GA) occurs in many plants. The adverse effects of GA are seldom cited. GA (6-14 µM) provoked the hemorrhagic liposis of the cervical muscles and intracranial hemorrhage. The cause of these pathological events and the method for prevention are still lacking. Using the chicken embryo model and some selected nutraceutics such as folate, glutathione (GSH), N-acetylcysteine, and vitamin E (Vit E), we carried out this study. Results revealed that the action mechanism of GA involved (i) inducing hypoxia with upregulated gene hif-1α and downregulated ratio vegf-r2/vegf-a, leading to dys-vascularization and myopathy; (ii) impairing cytochrome c oxidase; (iii) stimulating creatine kinase and lactate dehydrogenase release; (iv) eliciting carnitine accumulation and liposis via downregulating gene CPT1; (v) suppressing superoxide dismutase and stimulating NO, H2O2, and malondialdehyde; and (vi) depleting erythrocytic and tissue GSH, resulting in hemorrhage. When both Vit E and GSH were applied to the day 1 chicks, a better alleviation effect was revealed. Conclusively, GA potentially exhibits adverse effect by eliciting hemorrhagic liposis of cervical muscles and cerebral hemorrhage. Supplementation with GSH, Vit E, and N-acetylcysteine is able to ameliorate these adverse effects, warranting the importance of restricting the clinical phytotherapeutic doses of GA and related compounds.

Exercise ameliorates renal cell apoptosis in chronic kidney disease by intervening in the intrinsic and the extrinsic apoptotic pathways in a rat model.

We hypothesized that doxorubicin (DR) induced chronic kidney disease (CKD) could trigger the intrinsic and the extrinsic renal cell apoptotic pathways, while treadmill exercise could help prevent adverse effects. Male Sprague-Dawley rats were subjected to treadmill running exercise at a speed of 30 m/min, 30 or 60 min/day, 3 times per week, for a total period of 11 weeks. The physiological and biochemical parameters were seen substantially improved (DR-CKD control, 30 min, 60 min exercise): the ratio of kidney weight/body weight (0.89, 0.74, and 0.72); the WBC (1.35, 1.08, and 1.42 × 10(4) cells/ µ L); RBC (5.30, 6.38, and 6.26 × 10(6) cells/ µ L); the platelet count (15.1, 12.8, and 11.3 × 10(5)/ µ L); serum cholesterol (659, 360, and 75 mg/dL); serum triglyceride (542, 263, and 211 mg/dL); BUN (37, 25, and 22 mg/dL). Bcl-2 and intramitochondrial cytochrome c were upregulated, while the levels of Bax, SOD, MDA, cleaved caspases 9, 3, 8, 12, and calpain were all downregulated in DRCKD groups with exercise. CHOP (GADD153) and GRP78 were totally unaffected. FAS (CD95) was only slightly suppressed in the 60 min exercise DRCKD group. Conclusively, exercise can ameliorate CKD through the regulation of the intrinsic and extrinsic apoptosis pathways. The 60 min exercise yields more beneficial effect than the 30 min counterpart.

Unique bioactive polyphenolic profile of guava (Psidium guajava) budding leaf tea is related to plant biochemistry of budding leaves in early dawn.

BACKGROUND: Guava leaf tea (GLT), exhibiting a diversity of medicinal bioactivities, has become a popularly consumed daily beverage. To improve the product quality, a new process was recommended to the Ser-Tou Farmers' Association (SFA), who began field production in 2005. The new process comprised simplified steps: one bud-two leaves were plucked at 3:00-6:00 am, in the early dawn period, followed by withering at ambient temperature (25-28 °C), rolling at 50 °C for 50-70 min, with or without fermentation, then drying at 45-50 °C for 70-90 min, and finally sorted. RESULTS: The product manufactured by this new process (named herein GLTSF) exhibited higher contents (in mg g(-1), based on dry ethyl acetate fraction/methanolic extract) of polyphenolics (417.9 ± 12.3) and flavonoids (452.5 ± 32.3) containing a compositional profile much simpler than previously found: total quercetins (190.3 ± 9.1), total myricetin (3.3 ± 0.9), total catechins (36.4 ± 5.3), gallic acid (8.8 ± 0.6), ellagic acid (39.1 ± 6.4) and tannins (2.5 ± 9.1). CONCLUSION: We have successfully developed a new process for manufacturing GLTSF with a unique polyphenolic profile. Such characteristic compositional distribution can be ascribed to the right harvesting hour in the early dawn and appropriate treatment process at low temperature, avoiding direct sunlight.

Selected nutraceutic screening by therapeutic effects on doxorubicin-induced chronic kidney disease.

SCOPE: The number of patients with chronic kidney disease (CKD) are increasing. Interventions such as controlling hypertension and specific pharmacologic options are recommended. Some nutraceutics may have benefits in this regard. METHODS AND RESULTS: Naringenin (a flavanon), catechin (a flavanol), and quercetin (a flavonol) and rutin (a flavonol rutinoside) were tried on CKD in a Sprague Dawley rat model. Results indicated quercetin to be the most effective therapeutic candidate with respect to renal edema, hypertension, serum creatinine, hematocrit, cardiopathy, aorta calcification, glomerular amyloidosis, erythrocyte depletion in bone marrow, collagen deposition, expressions of TNF-α, cleaved caspase-3, IκBα, PPARα, and serum insulin. But quercetin was only partially effective in restoring glomerular filtration rate, albuminuria, serum cholesterol, triglyceride, blood urea nitrogen (BUN), uric acid, malondialdehyde, superoxide dismutase; urinary BUN and urinary creatinine. As for signaling, quercetin was completely effective in alleviating the cleaved caspase-3, being only partially effective in suppressing Bax and Bad, restoring Bcl-2, and rescuing DNA damage. CONCLUSION: The CKD status cannot to be ameliorated by naringenin, rutin, and catechin. Comparatively, quercetin may be a better therapeutic candidate.

Ferulic acid is nephrodamaging while gallic acid is renal protective in long term treatment of chronic kidney disease.

BACKGROUNDS & AIMS: The long term therapeutic effect of ferulic acid (FA) and gallic acid (GA) in treatment of chronic kidney disease (CKD) has been lacking. METHODS: Doxorubicin (DR, Adriamycin)-induced CKD rat model was established for this study. RESULTS: DR significantly reduced levels of serum albumin, GOT, GPT, RBC, TNF-α, and urinary creatinine and elevated serum cholesterol, TG, BUN, creatinine, uric acid, WBC, platelet count, and IL-6. In DRCKD rats, FA and GA significantly increased kidney weight and glomerular volume. FA reduced glomerular filtration rate but GA did not. FA enhanced more collagen deposition than GA in renal cortex and glomeruli. Both FA and GA showed crucial hyperlipidemic activity. The inhibitory effects of FA and GA on MMP-2 were very comparable. GA suppressed MMP-2 more effectively than FA in DRCKD rats. Both FA and GA induced SOD elevation and MDA elimination. In DRCKD rats, Western blot analysis indicated that FA further up-regulated CD34, α-SMA, tissue pDGFR, p-PDGFR, and TGF-ß; and down-regulated p-PI3K, and p-Akt. Since both PDGF-BB and TGF-ß are considered to induce kidney prefibrosis stage, GA was proved to be more beneficial in this regard. CONCLUSIONS: GA tends to protect the CKD while FA is not recommended for the long term CKD therapy.

The Aqueous Soluble Polyphenolic Fraction of Psidium guajava Leaves Exhibits Potent Anti-Angiogenesis and Anti-Migration Actions on DU145 Cells.

The aqueous extract of Psidium guajava budding leaves (PE) bears an extremely high content of polyphenolic and isoflavonoids. Whether it could be used as an anti-tumor chemopreventive in view of anti-angiogenesis and anti-migration, we performed the assay methods including the MTT assay to examine the cell viability; the ELISA assay to test the expressions of VEGF, IL-6 and IL-8; the western blot analysis to detect TIMP-2; the gelatinolytic zymography to follow the expression of MMPs; the wound scratch assay to examine the migration capability; and the chicken chorioallantoic membrane assay to detect the suppressive angiogenesis. Results indicated that the IC50 of PE for DU145 cells was ∼0.57 mg ml(-1). In addition, PE effectively inhibited the expressions of VEGF, IL-6 and IL-8 cytokines, and MMP-2 and MMP-9, and simultaneously activated TIMP-2 and suppressed the cell migration and the angiogenesis. Conclusively, PE potentially possesses a strong anti-DU145 effect. Thus, clinically it owns the potential to be used as an effective adjuvant anti-cancer chemopreventive.

Structural Characteristics and Antioxidative Capability of the Soluble Polysaccharides Present in Dictyophora indusiata (Vent. Ex Pers.) Fish Phallaceae.

Dictyophora indusiata (Vent. ex Pers.) Fish Phallaceae (Chinese name Zhu-Sun, the bamboo fungi) has been used as a medicinal mushroom to treat many inflammatory, gastric and neural diseases since 618 AD in China. We hypothesize that the soluble polysaccharides (SP) present in D. indusiata and their monosaccharide profiles can act as an important role affecting the antioxidative capability, which in turn would influence the biological activity involving anti-inflammatory, immune enhancing and anticancer. We obtained six SP fractions and designated them as D1, a galactoglucan; D2, a galactan; D3, the isoelectrically precipitated riboglucan from 2% NaOH; D4, a myoinositol; D5 and D6, the mannogalactans. The total SP accounted for 37.44% w/w, their molecular weight (MW) ranged within 801-4656 kDa. D3, having the smallest MW 801 kDa, exhibited the most potent scavenging effect against the α,α-diphenyl-ß-picrylhydrazyl, •OH(-), and •O(2) (-) radicals, yielding IC(50) values 0.11, 1.02 and 0.64 mg mL(-1), respectively. Thus we have confirmed our hypothesis that the bioactivity of D. indusiata is related in majority, if not entirely, to its soluble polysaccharide type regarding the MW and monosaccharide profiles.

The polyphenolics in the aqueous extract of Psidium guajava kinetically reveal an inhibition model on LDL glycation.

Guava [Psidium guajava L. (Myrtaceae)] budding leaf extract (PE) has shown tremendous bioactivities. Previously, we found seven major compounds in PE, i.e., gallic acid, catechin, epicatechin, rutin, quercetin, naringenin, and kaempferol. PE showed a potentially active antiglycative effect in an LDL (low density lipoprotein) mimic biomodel, which can be attributed to its large content of polyphenolics. The glycation and antiglycative reactions showed characteristic distinct four-phase kinetic patterns. In the presence of PE, the kinetic coefficients were 0.000438, 0.000060, 0.000, and -0.0001354 ABS-mL/mg-min, respectively, for phases 1 to 4. Computer simulation evidenced the dose-dependent inhibition model. Conclusively, PE contains a large amount of polyphenolics, whose antiglycative bioactivity fits the inhibition model.

Modeling of the in vivo kinetics of antioxidants delineates suitable parameters for selecting potential antioxidant adjuvants for cancer therapy.

To find in vivo behaviors of an antioxidant when used as an adjuvant cancer therapy, a more detailed integrated pharmacokinetic scheme is needed. Major reaction parameters associated with the sequential routes from ingestion to decay of an antioxidant were used in mathematical analysis, which included absorption rate coefficient k(a), quenching rate coefficient of the antioxidant k(q1) and tissue quenching rate coefficient k(r). The model was then treated with computer simulation using cited decay rate coefficients and some assumed parameters. When intestinal absorption rate coefficient k(a) becomes larger, retention time of antioxidant in plasma would be prolonged. moreover, k(a) had no effect on either quenching ability of antioxidants or tissue recovering capability. in quenching plasma ROS, the larger the quenching coefficient k(q1), the shorter peak- and the life-times would be for the secondary free radicals that are formed in primary quenching. Conclusively, it is suggestive to prescribe an antioxidant therapy with an appropriate values of k(a) and larger values of k(q1).

Action mechanism and signal pathways of Psidium guajava L. aqueous extract in killing prostate cancer LNCaP cells.

Aqueous extract of Psidium guajava L. budding leaves (PE) has been shown to possess anti-prostate cancer activity in a cell line model. We examined whether its bioactivity could be conserved either in the presence or the absence of synthetic androgen R1881. In both cases, PE was shown to inhibit LNCaP cell proliferation and down-regulate expressions of androgen receptor (AR) and prostate specific antigen (PSA). The cytotoxicity of PE was shown by enhanced LDH release in LNCaP cells. The flow cytometry analysis revealed cell cycle arrests at G(0)/G(1) phase with huge amount of apoptotic LNCaP cells after treatment with PE for 48 h in a dose-responsive manner, which was also confirmed by TUNEL assay. From the results of decreased Bcl-2/Bax ratio, inactivation of phosphor-Akt, activation of phosphor-p38, phospho-Erk1/phospho-Erk2, the molecular action mechanism of PE to induce apoptosis in LNCaP cells was elucidated. Compatible with the in vitro study findings, treatment with PE (1.5 mg/mouse/day) significantly diminished both the PSA serum levels and tumor size in a xenograft mouse tumor model. Conclusively, PE is a promising anti-androgen-sensative prostate cancer agent.

Anisotropic diffusion deviates chicken embryo chorioallantoic membrane assay (CAM) to reflect inherent therapeutic behaviors.

Chorioallantoic membrane assay (CAM) has become a widely used tool for determination of anti-angiogenesis capability of many drugs including herbal extracts. Because varying results in same set of chicken embryos are often encountered, we developed the complex diffusion model that combined the Fick's second diffusion law, chemical-protein interaction (or binding) to explain the diffusion- or kinetic-limiting phenomena in egg white when performing CAM. In addition, we performed diffusion studies in egg white with Color Blue No. 1, Evans Blue, Color Red No. 40, and the aqueous extract of Psidium guajava budding leaves (PE) to support our model. Under same conditions, the diffusion coefficients of Blue No. 1, Evans Blue, Red No. 40, and PE were (2.0-2.8)x10(-9), (0.89-31)x10(-9), (2.8-12)x10(-9), and (7.0-21)x10(-9)m(2)s(-1), respectively, depending upon the distance diffused. Whilst at the interface of egg white and embryo (egg yolk), a site about 1cm apart from the aeration sac, the percent concentration reached only 10.5, 3.0, 3.6, and 2.2% of the original applied medicine, respectively. We conclude that CAM could only serve as a preliminary screening tool for angiogenesis, because the anisotropic diffusion in egg white affects greatly the effective dosages of medicines tested.

Relevance of phenolic diterpene constituents to antioxidant activity of supercritical CO(2) extract from the leaves of rosemary.

Isolation of phenolic diterpene constituents from the freeze-dried leaves of Rosmarinus officinalis has been obtained by supercritical extraction with carbon dioxide. To determine the ideal conditions for the maximum yield of extract, nine different conditions using three levels of pressures (3000, 4000 and 5000 psi) in combination with three temperatures at 40, 60 and 80 degrees C, respectively, in combination with the analyses of the corresponding antioxidant activities and constituents which existed in extracts has been investigated. The antioxidant activity of each obtained extract was determined by using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals test. GC/MS method was used as an alternative to conventional HPLC method for the determination of the principal antioxidant constituents in extract, including phenolic diterpenes carnosic acid (CA) and carnosol (CAL). The confirmation of CA and CAL in extract was forward performed by subjecting HPLC isolates from extract into an ion trap mass spectrometer through an electrospray ionization (ESI) interface for MS/MS analysis. These results indicate that an ideal extraction process was obtained at 5000 psi and 80 degrees C with an extraction yield of 4.27% (w/w) and rich in phenolic antioxidants CA and CAL as contents of 35.23 and 0.46 mg g(-1) in extract, respectively.

Molecular action mechanism against apoptosis by aqueous extract from guava budding leaves elucidated with human umbilical vein endothelial cell (HUVEC) model.

Chronic cardiovascular and neurodegenerative complications induced by hyperglycemia have been considered to be associated most relevantly with endothelial cell damages (ECD). The protective effects of the aqueous extract of Psidium guajava L. budding leaves (PE) on the ECD in human umbilical vein endothelial cell (HUVEC) model were investigated. Results revealed that glyoxal (GO) and methylglyoxal (MGO) resulting from the glycative and autoxidative reactions of the high blood sugar glucose (G) evoked a huge production of ROS and NO, which in turn increased the production of peroxynitrite, combined with the activation of the nuclear factor kappaB (NFkappaB), leading to cell apoptosis. High plasma glucose activated p38-MAPK, and high GO increased the expressions of p38-MAPK and JNK-MAPK, whereas high MGO levels induced the activity of ERK-MAPK. Glucose and dicarbonyl compounds were all found to be good inducers of intracellular PKCs, which together with MAPK acted as the upstream triggering factor to activate NFkappaB. Conclusively, high plasma glucose together with dicarbonyl compounds can trigger the signaling pathways of MAPK and PKC and induce cell apoptosis through ROS and peroxynitrite stimulation and finally by NFkappaB activation. Such effects of PE were ascribed to its high plant polyphenolic (PPP) contents, the latter being potent ROS inhibitors capable of blocking the glycation of proteins, which otherwise could have brought forth severe detrimental effects to the cells.

Brain derived metastatic prostate cancer DU-145 cells are effectively inhibited in vitro by guava (Psidium gujava L.) leaf extracts.

The aqueous extract of Psidium guajava L. (PE) inhibited the cancer cell DU-145 in a dose- and time-dependent manner. At 1.0 mg/mL, PE reduced the viability of PCa DU-145 (the androgen independent PCa cells) to 36.1 and 3.59%, respectively after 48 h and 72 h of incubations. The absolute cell viability suppressing capability (VSC)(AC) could reach 262.5 cells-mL-h/mg on exposure to PE for 72 h, corresponding to the safe ranges, i.e. the percent viability suppressing rates (PVSR) of 2.72 and 2.41 folds for DU-145 comparing to PZ-HPV-7 cells when treated with PE at 0.5 and 1.0 mg/mL respectively for 72 h. In addition, the colony forming capability of DU-145 cells was apparently lowered. The suppressing rates of which reached 8.09 and 5.96 colony/mg/day for D-145 and PZ-HPV-7 cells, respectively within the concentration range of PE at 0.1 asymptotically equal to 0.25 mg/mL. Cell cycle arrests at G0/G1 phase in both cells were observed by TUNEL assay and flow cytometric analysis, yet more prominently evident in DU-145. In addition, suppression of the matrix metalloproteinases MMP-2 and MMP-9, and the upregulation of active caspase-3 at 0.10 to 1.0 mg/mL in DU-145 were also effected in a dose-dependent manner by PE at 0.25 to 1.0 mg/mL, implicating a potent anti-metastasis power of PE. Conclusively, we ascribe the anticancer activity of PE to its extraordinarily high polyphenolic (165.61 +/- 10.39 mg/g) and flavonoid (82.85 +/- 0.22 mg/g) contents. Furthermore, PE might be useful for treatment of brain derived metastatic cancers such as DU-145, acting simultaneously as both a chemopreventive and a chemotherapeutic.

Low-density lipoprotein, collagen, and thrombin models reveal that Rosemarinus officinalis L. exhibits potent antiglycative effects.

Using the low-density lipoprotein (LDL), collagen, and thrombin models, we report here that the rosemary extracts (REs), either the aqueous (REw) or the acetonic (REA), all possessed many antiglycation-related features, and the effective concentrations required were as follows: 0.1 mg/mL for suppressing the relative electrophoretic mobility, 1.3 microg/mL for anticonjugated diene induction, 0.5 mg/mL for inhibition of thiobarbituric acid reactive substances production, 0.1 mg/mL for AGEs (advanced glycation end products) formation, 0.1 mg/mL to block glucose incorporation, and 0.05 mg/mL as an effective anti-antithrombin III. Using high-performance liquid chromatography/mass spectrometry, we identified five major constituents among eight major peaks, including rosmarinic acid, carnosol, 12-methoxycarnosic acid, carnosic acid, and methyl carnosate. In the LDL model, REA was proven to be more efficient than REw; yet, the reverse is true for the collagen and the thrombin III models, the reason of which was ascribed to the higher lipid-soluble antioxidant content (such as rosmarinic acid, carnosol, carnosic acid, 12-methoxycarnosic acid and methyl carnosate) in REA than in REw and the different surface lipid characteristics between LDL and collagen; although to act as anti-AGEs, both extracts were comparable. To assist the evidence, a larger 2,2-diphenyl-1-picrylhydrazyl radical scavenging capability with less total polyphenolic content was found in REA. We conclude that rosemary is an excellent multifunctional therapeutic herb; by looking at its potential potent antiglycative bioactivity, it may become a good adjuvant medicine for the prevention and treatment of diabetic, cardiovascular, and other neurodegenerative diseases.

Kinetic analysis on the sensitivity of glucose- or glyoxal-induced LDL glycation to the inhibitory effect of Psidium guajava extract in a physiomimic system.

Experimentation with a physiomimic system and kinetic analysis exhibited four distinct reaction phases in LDL glycation despite of the type of inducer: glucose or glyoxal. LDL glycation was more sensitive to a status of hyperglycemia (such as 400 mg glucose/100 mL) as evidenced by the reaction order of 0.53. Glucose reacted intensively in the Initial Phase (reaction period 0-2h) which was identified to result from a parallel mechanism involving both the direct Schiff's product formation and the auto-oxidative cleavages. In contrast, a physiological level of glyoxal revealed merely a reaction order of only 0.09, implicitly indicating a far less sensitive glycation which can be attributed to a mechanism proceeding simply through a molecular Schiff's reaction. On treatment with Psidium guajava L. aqueous extract (PE) (0.01-0.625 mg/mL), a rather unique and significant inhibitory characteristic on LDL glycation was observed with a dose-dependent manner. We attributed such an effect of PE to its distinct abundance of polyphenolic content (165.61+/-10.39 mggallic acid equivalent (GAE)/g). Conclusively, PE is an excellent anti-LDL glycative agent whose potential therapeutic uses can be extended to the prevention of a variety of cardiovascular and neurodegenerative diseases associated with glycations.

Inhibitory effect of some selected nutraceutic herbs on LDL glycation induced by glucose and glyoxal.

Anti-LDL glycative agents were investigated using aqueous extracts of Psidium guajava L. (PE), Toona sinensis Roem. (TE), Momordica charantia L. (ME) and Graptopetalum paragugayene E. Walther (GE). Concentrations of extracts 0.01-0.625 mg/mL, low density lipoprotein (LDL; 100 microg protein/mL) and inducers glucose (400 mM) and glyoxal (2.5 mM) were incubated at 37 degrees C. Evaluation parameters involved the thiobarbituric acid reactive substances (TBARS), conjugated dienes (CD), relative electrophoretic mobility (REM), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capability and total polyphenolic content. Results for anti-TBARS efficiency (in%) were PE (75.77), TE (75.10), ME (68.81) and GE (19.81) at 0.5 mg/mL, respectively, when induced by glucose; 36.68, 35.60, 32.62 and inactive, respectively, by glyoxal. The lag times for CD formation (in min) were: 289 and 125 by PE and TE, respectively, comparing to the control (45). REM was 1.6 with respect to PE (0.1 mg/mL) compared to the control (4.2). PE at 0.01 mg/mL effectively inhibited with 63.45% efficiency on AGEs induced by glucose. We conclude that PE virtually is a potent antiglycative agent, which can be of great value in the preventive glycation-associated cardiovascular and neurodegenerative diseases.