Nanteetip Limpeanchob

Neuroprotective effect of Bacopa monnieri on beta-amyloid-induced cell death in primary cortical culture.

AIM OF THE STUDY Bacopa monnieri (Brahmi) is extensively used in traditional Indian medicine as a nerve tonic and thought to improve memory. To examine the neuroprotective effects of Brahmi extract, we tested its protection against the beta-amyloid protein (25-35) and glutamate-induced neurotoxicity in primary cortical cultured neurons. MATERIALS AND METHODS Neuroprotective effects were determined by measuring neuronal cell viability following beta-amyloid and glutamate treatment with and without Brahmi extract. Mechanisms of neuroprotection were evaluated by monitoring cellular oxidative stress and acetylcholinesterase activity. RESULTS Our result demonstrated that Brahmi extract protected neurons from beta-amyloid-induced cell death, but not glutamate-induced excitotoxicity. This neuroprotection was possibly due to its ability to suppress cellular acetylcholinesterase activity but not the inhibition of glutamate-mediated toxicity. In addition, culture medium containing Brahmi extract appeared to promote cell survival compared to neuronal cells growing in regular culture medium. Further study showed that Brahmi-treated neurons expressed lower level of reactive oxygen species suggesting that Brahmi restrained intracellular oxidative stress which in turn prolonged the lifespan of the culture neurons. Brahmi extract also exhibited both reducing and lipid peroxidation inhibitory activities. CONCLUSIONS From this study, the mode of action of neuroprotective effects of Brahmi appeared to be the results of its antioxidant to suppress neuronal oxidative stress and the acetylcholinesterase inhibitory activities. Therefore, treating patients with Brahmi extract may be an alternative direction for ameliorating neurodegenerative disorders associated with the overwhelming oxidative stress as well as Alzheimers disease.

Screening for phosphodiesterase inhibitory activity of Thai medicinal plants

INTRODUCTION Phosphodiesterases (PDEs) are a group of enzymes that have powerful effects on cellular signaling because they regulate the second messenger, cAMP or cGMP. PDE inhibitors have been used for treatment of many indications such as cardiovascular diseases, chronic obstructive pulmonary diseases, erectile dysfunction and pulmonary hypertension. THE AIM OF THE STUDY The aim of the study was to search for sources of PDE inhibitors from Thai biodiversity. MATERIALS AND METHODS Some Thai medicinal plants used as aphrodisiac and neurotonic agents together with plants from Leguminosae collected from the North of Thailand were screened for PDE inhibitory activity using a radioassay. RESULTS Seven from nineteen aphrodisiac and neurotonic plants as well as three from twelve Leguminosae plants showed potent PDEs inhibitory activity. The concentrations that could inhibit 50% PDE activity (IC(50)) of the active extracts were determined in comparison to the standard inhibitor, 3-isobutyl-1-methylxanthine (IBMX). Betula alnoides, Hiptage benghalensis, Leea indica and Senna surrattensis showed IC(50) values in the range of microgram per milliliter while IBMX standard showed an IC(50) value of 0.68+/-0.13 microg/ml. CONCLUSION Thai biodiversity was the great sources of PDE inhibitors.

Effects of silk sericin on the proliferation and apoptosis of colon cancer cells

Sericin is a silk protein woven from silkworm cocoons (Bombyx mori). In animal model, sericin has been reported to have anti-tumoral action against colon cancer. The mechanisms underlying the activity of sericin against cancer cells are not fully understood. The present study investigated the effects of sericin on human colorectal cancer SW480 cells compared to normal colonic mucosal FHC cells. Since the size of the sericin protein may be important for its activity, two ranges of molecular weight were tested. Sericin was found to decrease SW480 and FHC cell viability. The small sericin had higher anti-proliferative effects than that of the large sericin in both cell types. Increased apoptosis of SW480 cells is associated with increased caspase-3 activity and decreased Bcl-2 expression. The anti-proliferative effect of sericin was accompanied by cell cycle arrest at the S phase. Thus, sericin reduced SW480 cell viability by inducing cell apoptosis via caspase-3 activation and down-regulation of Bcl-2 expression. The present study provides scientific data that support the protective effect of silk sericin against cancer cells of the colon and suggests that this protein may have significant health benefits and could potentially be developed as a dietary supplement for colon cancer prevention.

Sericin reduces serum cholesterol in rats and cholesterol uptake into Caco-2 cells.

A cholesterol lowering effect of sericin was investigated both in vivo and in vitro. Rats were dosed with cholesterol with and without sericin for 14 days. Non-high-density lipoprotein (HDL) and total serum cholesterols were reduced in rats fed high-cholesterol diet with all three tested doses of sericin (10, 100, and 1000 mg kg(-1) day(-1)). The potential mechanism of actions was determined by measuring the uptake of radiolabeled cholesterol into differentiated Caco-2 cells and cholesterol solubility in mixed lipid micelles. Concentration of sericin as low as 25 and 50 μg/mL inhibited 30% of cholesterol uptake into Caco-2 cells whereas no effect was found at higher concentration. Cholesterol micellar solubility was reduced in the presence of sericin. This study suggests the cholesterol lowering effect of sericin results from its inhibition of cholesterol absorption in intestinal cells and its reduction of cholesterol solubility in lipid micelles.

Anti-androgenic effect of sesquiterpenes isolated from the rhizomes of Curcuma aeruginosa Roxb.

Six sesquiterpenes: germacrone (1), zederone (2), dehydrocurdione (3), curcumenol (4), zedoarondiol (5) and isocurcumenol (6) were isolated from rhizomes of Curcuma aeruginosa Roxb. (Zingiberaceae). They inhibited 5α-reductase which converts testosterone to dihydrotestosterone (DHT). Germacrone (1) was the most potent (IC(50)=0.42±0.05 mg/mL). Compound 1 was anti-androgenic in LNCaP cells when proliferation was testosterone-induced. The growth of flank gland of male Syrian hamsters is dependent on circulating androgen and when maintained with testosterone, 1 (3, 30, 100μg) inhibited growth but was ineffective against DHT. The similar activity profile was observed on the 5α-reductase inhibitor, finasteride (100 μg) treatment group. The androgen receptor binding assay showed that 1 did not bind to the androgen receptor. In conclusion, 1 showed anti-androgenic effect on in vitro and in vivo assays. One of the possible mechanisms was inhibition 5α-reductase activity. Thus, 1 is a potential lead compound for treatment of androgen-dependent disorders.

Bisindole alkaloids and secoiridoids from Alstonia macrophylla Wall. ex G. Don

The ethanolic extract from stems of a Thai medicinal plant, Alstonia macrophylla Wall. ex G. Don (Apocynaceae) showed a significant inhibitory effect on acetylcholinesterase (AChE) determined by using Ellman assay. Four compounds i.e., a bisindole alkaloid, macralstonine (1), a new bisindole alkaloid, thungfaine (2), a secoiridoid glycoside, sweroside (3) and a new secoiridoid glycoside, naresuanoside (4) were isolated. Compound 4 showed moderate AChE and butyrylcholinesterase (BChE) inhibitory effects. Interestingly, compound 4 inhibited cell growth on human androgen-sensitive prostate cancer cell line (LNCaP) but no effect on viability of human foreskin fibroblast cells (HF).

Silk lutein extract and its combination with vitamin E reduce UVB-mediated oxidative damage to retinal pigment epithelial cells

Increased exposure to solar ultraviolet B (UVB) radiation may promote age related macular degeneration (AMD). Lutein can protect retinal pigment epithelial (RPE) cells from various oxidative insults but its direct protection against UVB has not been reported. This study aimed to demonstrate protective effects of silk lutein extract against UVB-induced oxidative damage to RPE cells and compared with standard lutein and Trolox, a vitamin E analog. ARPE-19 cells were treated with luteins with and without Trolox prior to UVB exposure. Cell viability and apoptosis were determined by trypan blue staining and caspase-3 activity, respectively. Oxidative damage was evaluated by measuring intracellular reactive oxygen species (ROS), lipid peroxidation, and activities of antioxidant enzymes (superoxide dismutase, glutathione peroxidase and catalase). Levels of lutein remained in culture medium was determined by HPLC. Both luteins reduced cellular ROS levels and lipid peroxidation mediated by UVB, and subsequently increased cell viability and reduced apoptosis. They also restored activities of most tested antioxidant enzymes. Enhancement of lutein antioxidant efficacy was observed in the presence of Trolox. In all these effects, the two lutein preparations had similar effectivenesses. In cell free media, Trolox enhanced the protective effect of lutein probably by reducing its degradation and repairing the oxidized derivatives. Yellow silk cocoon is a potential candidate of lutein for further development as dietary supplement for the prevention of AMD.

Potential mechanisms of hypocholesterolaemic effect of Thai spices/dietary extracts

Several Thai spices/dietary ingredients were previously shown to have hypocholesterolaemic effects. These studies were mostly conducted in animal models in which the mechanisms of action were not yet well-established. Therefore, this study aimed to investigate the potential mechanism of hypocholesterolaemic action of 12 selected plants, namely Hibiscus sabdariffa L., Moringa oleifera Lam., Cucurbita moschata Duchesne, Ananas comosus (L.) Merr., Zingiber officinale, Morus alba L., Camellia sinensis (L.) Kuntze, Piper nigrum L., Alpinia galanga (L.) Willd., Curcuma zedoaria Rose, Bacopa monnieri (L.) Wettst. and Piper retrofractum Vahl., widely used as spices and ingredients in various types of Thai food. The extract of P. nigrum at 100 µg mL−1 was found to be the most effective cholesterol uptake inhibitor whereas those of A. galanga and C. sinensis effectively inhibited pancreatic lipase activity with IC50 values of 8.99 ± 3.41 and 12.36 ± 1.23 µg mL−1, respectively. The potency of extracts from H. sabdariffa, M. oleifera and C. moschata at 100 µg mL−1 were found to be similar to 0.4 µg mL−1 pravastatin in inhibiting HMG-CoA reductase and possibly reduced cholesterol biosynthesis. This study also demonstrated that several of the tested plants possessed multiple sites of action that were possibly responsible for their cholesterol-lowering effect in the in vivo model.

Protective effect of silk lutein on ultraviolet B-irradiated human keratinocytes

Carotenoids are efficient antioxidants that are of great importance for human health. Lutein and zeaxanthin are carotinoids present in high concentrations in the human retina which are involved in the photoprotection of the human eye. Lutein may also protect the skin from ultraviolet (UV)-induced damage. The present study investigated the protective effect of lutein extracted from yellow silk cocoons of Bombyx mori on human keratinocytes against UVB irradiation. A human keratinocyte cell line and primary human keratinocytes were used to investigate the UVB protection effects of silk lutein and plant lutein. Silk lutein showed no cytotoxicity to keratinocytes. Treatment with silk lutein prior to UVB irradiation enhanced cell viability and cell proliferation, and reduced cell apoptosis. The protective effects of silk lutein may be superior to those of plant lutein. Silk lutein may have a benefit for protection of keratinocytes against UVB-irradiation.

Oxidized low density lipoprotein increases acetylcholinesterase activity correlating with reactive oxygen species production.

Hyperlipidemia, low density lipoproteins (LDL) and their oxidized forms, and oxidative stress are suspected to be a key combination in the onset of AD and acetylcholinesterase (AChE) plays a part in this pathology. The present study aimed to link these parameters using differentiated SH-SY5Y human neuroblastoma cells in culture. Both mildly and fully oxidized human LDL (mox- and fox-LDL), but not native (non-oxidized) LDL were cytotoxic in dose- and time-dependent patterns and this was accompanied by an increased production of intracellular reactive oxygen species (ROS). Oxidized LDL (10-200 μg/mL) augmented AChE activity after 4 and 24h treatments, respectively while the native LDL was without effect. The increased AChE with oxidized LDLs was accompanied by a proportionate increase in intracellular ROS formation (R=0.904). These findings support the notion that oxidized LDLs are cytotoxic and that their action on AChE may reduce central cholinergic transmission in AD and affirm AChE as a continued rational for anticholinesterase therapy but in conjunction with antioxidant/antihyperlipidemic cotreatments.