Neuroprotective effects of Hu-Yi-Neng, a diet supplement, on SH-SY5Y human neuroblastoma cells.

OBJECTIVES: Oxidative stress is considered the potential risk to the development of dementia. Some medicines, vitamins, and diet supplements have been suggested to have possible benefits via the antioxidative effects to slow the decline of cognitive function in demented and non-demented individuals. However, few studies were conducted to examine their functions, especially in composite diet supplements. Hu-Yi-Neng is a composite diet supplement, including ginkgo biloba, extract of pine bark, phosphatidyl serine, docosahexaenoic acid, and folic acid, used extensively in Taiwan. Therefore, our aim is to investigate the potential protective effects of Hu-Yi-Neng on human neuron cells. MATERALS AND METHODS: H2O2-induced neuronal toxicity was characterized in SH-SY5Y human neuroblastoma cells by the decrease of cell viability using PrestoBlue™ assay and by the increase of intracellular reactive oxygen species (ROS) level using DCFH-DA (2', 7'-dichlorodihydrofluorescin diacetate) assays. HO-1 mRNA expression was detected by real-time PCR. Akt and Erk 1/2 proteins were detected by western blotting. RESULTS: Pretreatment with Hu-Yi-Neng significantly reversed the decrease in cell viability induced by H2O2 in SH-SY5Y cells. Furthermore, Hu-Yi-Neng dose-dependently suppressed the elevation of intracellular reactive oxygen species (ROS) level. Hu-Yi-Neng protected SH-SY5Y cells from oxidative stress may via the increase in mRNA expression of heme oxygenase-1 (HO-1), an antioxidant enzyme. In addition, Hu-Yi-Neng inhibited H2O2-induced phosphorylation of Akt kinase but further increased the phosphorylation of Erk 1/2. CONCLUSION: Our results suggest that Hu-Yi-Neng has protective effect against oxidative stress-induced neuron cell loss and it could be an ideal composite diet supplement for preventing neurodegenerative diseases.

Antioxidant, antioedema and analgesic activities of Andrographis paniculata extracts and their active constituent andrographolide.

Andrographis paniculata (AP), a popular ingredient of Oriental folk medicine, is commonly used for treating infection, inflammation, fever and diarrhoea. In this study, extracts prepared from cultivated AP and their active constituent andrographolide were evaluated for antioxidant, antioedema and analgesic activities. The results showed that the aqueous AP extract (AP-H2O) exhibited a greater antioxidant activity than the ethanol AP extract (AP-EtOH) in all model systems tested. At a concentration of 50 microg/mL, the free radical scavenging, xanthine oxidase inhibition and antilipid peroxidation activities for AP-H2O were 66.8%, 57.3% and 65.3%, respectively, and for AP-EtOH were 57.8%, 52.6% and 34.2%, respectively. At a dosage of 100 mg/kg, AP-H2O and andrographolide, but not AP-EtOH, showed antioedema and analgesic activities. In phytochemical analysis, AP-H2O showed a higher concentration of total flavanoid but a lower phenol content than AP-EtOH. In conclusion, AP-H2O was more potent than AP-EtOH in antioxidant activities. Furthermore, compared with andrographolide, AP-H2O as an extract also appears to possess potent antioedema and analgesic activities.

Supercritical carbon dioxide extract exhibits enhanced antioxidant and anti-inflammatory activities of Physalis peruviana.

Physalis peruviana L. (PP) is a medicinal herb widely used in folk medicine. In this study, supercritical carbon dioxide (SFE-CO2) method was employed to obtain three different PP extracts, namely SCEPP-0, SCEPP-4 and SCEPP-5. The total flavonoid and phenol concentrations, as well as antioxidant and anti-inflammatory activities of these extracts were analyzed and compared with aqueous and ethanolic PP extracts. Among all the extracts tested, SCEPP-5 demonstrated the highest total flavonoid (234.63+/-9.61 mg/g) and phenol (90.80+/-2.21 mg/g) contents. At concentrations 0.1-30 microg/ml, SCEPP-5 also demonstrated the strongest superoxide anion scavenging activity and xanthine oxidase inhibitory effect. At 30 microg/ml, SCEPP-5 significantly prevented lipopolysaccharide (LPS; 1 microg/ml)-induced cell cytotoxicity in murine macrophage (Raw 264.7) cells. At 10-50 microg/ml, it also significantly inhibited LPS-induced NO release and PGE2 formation in a dose-dependent pattern. SCEPP-5 at 30 microg/ml remarkably blocked the LPS induction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. Taken together, these results suggest that SCEPP-5, an extract of SFE-CO2, displayed the strongest antioxidant and anti-inflammatory activities as compared to other extracts. Its protection against LPS-induced inflammation could be through the inhibition of iNOS and COX-2 expression.

Evaluation of hepatoprotective activity of legumes.

Mung bean, adzuki bean, black bean and rice bean are foods and folk medicines of Taiwan. We evaluated the effects of various water extract concentrations (100, 500 and 1000 mg/kg body wt.) and silymarin (25 mg/kg body wt. on acetaminophen-induced liver injury by measuring serum glutamate-oxalate-transaminase (sGOT) and serum glutamate-pyruvate-transaminase (sGPT) activities in rats. The results showed that the sGOT and the sGPT activities, increased by APAP, were decreased significantly (P < 0.05) through treatment with inceasing amounts up to 1000 mg/kg body wt. of the exracts. In particular, the mung bean aqueous extract showed the best hepatoprotective effect on APAP-induced hepatotoxicity. The pathological changes of liver injury caused by APAP improved by the treatment with all of the legume extracts, which were compared to silymarin as a standardized drug. In addition to these results, the extract of mung bean acted as a potential hepatoprotective agent in dietary supply.

Antiplatelet and antithrombotic effects of a combination of ticlopidine and ginkgo biloba ext (EGb 761).

The antiplatelet and antithrombotic effects of the oral combination treatment of ticlopidine and Ginkgo biloba extract (EGb 761) were studied in normal and thrombosis-induced rats. The ex vivo inhibitory effect on ADP-induced platelet aggregation of a small dose of ticlopidine (50 mg/kg/day) in combination with EGb 761 (40 mg/kg/day) was comparable to a larger dose of only ticlopidine (200 mg/kg/day). Bleeding time was also prolonged by 150%. Thrombus weight was also consistently decreased by a combination of ticlopidine and EGb 761 in an arterio-venous shunt model at two doses of ticlopidine (50 mg/kg) plus EGb 761 (20 mg/kg) and ticlopidine (50 mg/kg) plus EGb 761 (40 mg/kg). A combinatory treatment in acute thrombosis model in mice also showed a higher recovery than a single treatment.

Chemical and bioactive constituents from Zanthoxylum simulans.

Two new benzo[c]phenanthridine alkaloids, 6-methyldihydrochelerythrine [1] and 6-methylnorchelerythrine [2], together with 23 known compounds, were isolated from the root bark of Zanthoxylum simulans. Structures were elucidated by spectral analysis. Among them, the pyranoquinoline alkaloids, zanthosimuline [3], and huajiaosimuline [4], exhibited cytotoxic activity. In addition, compound 4 showed significant antiplatelet aggregation activity and induced terminal differentiation with cultured HL-60 cells.

Inhibition of collagen-induced platelet aggregation and adhesion by a pseudocyanide derivative of avicine isolated from Zanthoxylum integrifoliolum Merr.

Avicine pseudocyanide, a derivative of avicine isolated from Zanthoxylum integrifoliolum Merr., inhibited collagen-induced platelet aggregation and release reaction in a concentration-dependent manner. Trimucytin is a collagen-like snake venom protein isolated from Trimeresurus mucrosquamatus. Avicine pseudocyanide also inhibited trimucytin (1 microgram/mL)-induced platelet aggregation and release reaction concentration dependently. The IC50 values of avicine pseudocyanide on collagen (10 micrograms/mL)- and trimucytin (1 microgram/mL)-induced platelet aggregation were 47.3 +/- 4.1 and 62.5 +/- 5.6 microM, respectively. Avicine pseudocyanide at a concentration of 300 microM inhibited less than 30% of platelet aggregation induced by ADP (20 microM), AA (100 microM), U46619 (1 microM), PAF (2 ng/mL) and thrombin (0.1 U/mL). The concentration-response curve of collagen-induced platelet aggregation was shifted to the right by avicine pseudocyanide (20-100 microM) concentration dependently. The Schild plot showed that pA2 and pA10 values of avicine pseudocyanide were 4.8 and 4.3, respectively, with slope of -1.9. Avicine pseudocyanide also inhibited collagen (10 micrograms/mL)-induced aggregation of rabbit whole blood with an IC50 of 145 +/- 13 microM. Collagen-induced thromboxane B2 formation was also inhibited by avicine pseudocyanide in a concentration-dependent manner with a maximal effect at 100 microM. However, arachidonic acid (AA)-induced thromboxane B2 and prostaglandin D2 formations were only partially suppressed by a high concentration of avicine pseudocyanide (300 microM). Avicine pseudocyanide (100 microM) inhibited the [3H]inositol monophosphate formation and the rise of intracellular Ca2+ concentration caused by collagen but not those caused by AA, U46619, platelet-activating factor and thrombin. In the presence of prostaglandin E1, Mg(2+)-dependent platelet adhesion to collagen was inhibited by avicine pseudocyanide with an IC50 of 278 +/- 16 microM. These data indicate that avicine pseudocyanide is an inhibitor of collagen-induced platelet aggregation and platelet-collagen adhesion.

Determination of erythrocyte glutathione peroxidase activity and its reference range in Chinese adults.

A method to assess selenium status of the body by measuring glutathione peroxidase activity in erythrocytes was studied. Reaction was measured by continuous monitoring of the decrease of NADPH at 340 nm. The erythrocyte glutathione peroxidase activity was determined at 37 degrees C, pH 7.5 by using one of the substrates, t-butyl hydroperoxide, to initiate the reaction, and using glutathione reductase as the coupling enzyme. The Km of the enzyme for glutathione and t-butyl hydroperoxide were determined to be 1.8 mM and 238 microM, respectively. The enzyme was stable at -20 degrees C or -70 degrees C for at least 5 months, and for at least 2 months when stored at 4 degrees C. The within-run and between-run coefficients of variation for this method were 3.3-4.9% and 3.0-7.1%, respectively. The test was linear up to 100 U/g hemoglobin, and had a sensitivity of 0.002 delta A/min at 3 U/L. The reference range of erythrocyte glutathione peroxidase in Chinese adults was estimated to be 28.6-87.8 U/g hemoglobin (n = 84), without a significant difference in the results between males and females.

Antiplatelet effects of chelerythrine chloride isolated from Zanthoxylum simulans.

Chelerythrine chloride is an antiplatelet agent isolated from Zanthoxylum simulans. Aggregation and ATP release of washed rabbit platelets caused by ADP, arachidonic acid, PAF, collagen, ionophore A23187 and thrombin were inhibited by chelerythrine chloride. Less inhibition was observed in platelet-rich plasma. The thromboxane B2 formation of washed platelets caused by arachidonic acid, collagen, ionophore A23187 and thrombin was decreased by chelerythrine chloride. Phosphoinositides breakdown caused by collagen and PAF was completely inhibited by chelerythrine chloride, while that of thrombin was only partially suppressed. Chelerythrine chloride inhibited the intracellular calcium increase caused by arachidonic acid, PAF, collagen and thrombin in quin-2/AM-loaded platelets. The cyclic AMP level of washed platelets did not elevated by chelerythrine chloride. The antiplatelet effect of chelerythrine chloride was not dependent on the incubation time and the aggregability of platelets inhibited by chelerythrine chloride was easily recovered after sedimenting the platelets by centrifugation and then the platelet pellets were resuspended. Chelerythrine chloride did not cause any platelet lysis, since lactate dehydrogenase activity was not found in the supernatant. These data indicate that the inhibitory effect of chelerythrine chloride on rabbit platelet aggregation and release reaction is due to the inhibition on thromboxane formation and phosphoinositides breakdown.

Interleukin 1 and 2 production in homosexual men: a controlled trial of therafectin (SM-1213), a possible immunomodulator.

Patients with the acquired immunodeficiency syndrome (AIDS) are susceptible to a variety of opportunistic pathogens which require intact cellular immunity for control and eradication. We evaluated interleukin 1 and 2 production in 12 homosexual men without AIDS but with evidence of altered cell-mediated immunity and serologic evidence of infection with human T-cell lymphotrophic virus type III (HTLV-III), the etiologic agent of AIDS and found production of both factors diminished compared to heterosexual controls. Therafectin (SM-1213) is a new agent which selectively activates macrophages and stimulated interleukin 1 production in vitro. Therafectin was administered to these same 12 patients in a double-blind, placebo controlled trial. We failed to find any significant changes in their immunologic status including interleukin 1 or 2 production.

Effectiveness of intermediate-dose methotrexate and high-dose 5-fluorouracil as sequential combination chemotherapy in refractory breast cancer and as primary therapy in metastatic adenocarcinoma of the colon.

Many studies reported the pretreatment with methotrexate followed by 5-FU resulted in the greatest tumor cell killing. Our preliminary laboratory studies confirmed the possibility of drug synergism in the L1210 cell line, but not in human bone marrow cells. Thirteen patients with metastatic adenocarcinoma of the breast and seven patients with metastatic adenocarcinoma of the colon were treated with an innovative approach in which methotrexate preceded 5-FU administration in an attempt to cause drug synergism and prevent drug antagonism. All patients with breast cancer and two patients with colon cancer had been extensively pretreated with multiple drugs. Of the cancer patients so treated, three (23%) with breast cancer and two (28%) with colon cancer demonstrated objective tumor response. In addition to these patients, six (46%) with breast cancer and three (43%) with colon cancer demonstrated subjective improvement as manifested by total pain relief and reduction in CEA titer. The preliminary results reported in this study suggest that sequential utilization of intermediate doses of methotrexate, followed by high doses of 5-FU, is an effective combination chemotherapy for patients with breast and colon malignancies.