Suppression of colorectal cancer cell growth by combined treatment of 6-gingerol and γ-tocotrienol via alteration of multiple signalling pathways

Journal of Natural Medicines - Our previous study reported that combined treatment of γ-tocotrienol with 6-gingerol showed promising anticancer effects by synergistically inhibiting...     

Chlorella vulgaris Ameliorates Oxidative Stress and Improves the Muscle Regenerative Capacity of Young and Old Sprague-Dawley Rats

Muscle atrophy in ageing is a multifactorial degenerative process impacted by cellular ageing biology, which includes oxidative stress. Chlorella vulgaris is a coccoid green eukaryotic microalga rich in antioxidants. The aim of this study was to determine the effect of C. vulgaris in ameliorating oxidative stress, thus elucidating its mechanism in improving muscle mass, strength and function in young and old rats. Fifty-six male Sprague-Dawley (SD) rats aged 3 months (young) and 21 months (old) were divided into three groups: Group 1 (control) was given distilled water; Group 2 was treated with 150 mg/kg body weight (BW) of C. vulgaris; and Group 3 was treated with 300 mg/kg BW of C. vulgaris for three months. Grip and muscle strength and muscle integrity were determined on days 0, 30, 60, and 90 of treatment. Urine and blood were collected on days 0 and 90 of treatment for oxidative stress marker determination, while the gastrocnemius muscles were collected for muscle oxidative stress analysis. Increased grip strength of the front and hind paws was observed in young C. vulgaris-treated rats on days 30, 60, and 90 compared to the untreated control on the same days (p < 0.05). There was a significant increase in lean bone mineral content (BMC) in young rats treated with 300 mg/kg BW C. vulgaris compared to untreated rats on days 30 and 60. The fat mass was significantly decreased in young and old C. vulgaris-treated rats on day 90 compared to the untreated control. The total path was significantly increased for old rats treated with 300 mg/kg BW C. vulgaris on days 60 and 90 compared to day 0. Young and old C. vulgaris-treated rats demonstrated a significant decrease in urinary isoprostane F_2t and plasma creatine kinase-MM (CKMM) compared to the control on day 90. A significant decrease in malondialdehyde (MDA) and 4-hydroxyalkenal (HAE) levels were observed in young and old rats treated with C. vulgaris. C. vulgaris improved the muscle mass, strength, and function in young and old rats. This effect could be due to its potency in ameliorating oxidative stress in the skeletal muscle of young and old rats.     

Modulation of collagen synthesis and its gene expression in human skin fibroblasts by tocotrienol-rich fraction

Introduction

Skin aging may occur as a result of increased free radicals in the body. Vitamin E, the major chain-breaking antioxidant, prevents propagation of oxidative stress, especially in biological membranes. In this study, the molecular mechanism of tocotrienol-rich fraction (TRF) in preventing oxidative stress-induced skin aging was evaluated by determining the rate of total collagen synthesis and its gene expression in human skin fibroblasts.

Material and methods

Primary culture of human skin fibroblasts was derived from circumcision foreskin of 9 to 12 year-old boys. Fibroblast cells were divided into 5 different treatment groups: untreated control, hydrogen peroxide (H₂O₂)-induced oxidative stress (20 µM H₂O₂ exposure for 2 weeks), TRF treatment, and pre- and post-treatment of TRF to H₂O₂-induced oxidative stress.

Results

Our results showed that H₂O₂-induced oxidative stress decreased the rate of total collagen synthesis and down-regulated COL I and COL III in skin fibroblasts. Pre-treatment of TRF protected against H₂O₂-induced oxidative stress as shown by increase in total collagen synthesis and up-regulation of COL I and COL III (p<0.05) genes. However, similar protective effects against H₂O₂-induced oxidative stress were not observed in the post-treated fibroblasts.

Conclusions

Tocotrienol-rich fraction protects against H₂O₂-induced oxidative stress in human skin fibroblast culture by modulating the expression of COL I and COL III genes with concomitant increase in the rate of total collagen synthesis. These findings may indicate TRF protection against oxidative stress-induced skin aging.     

Hot water extract of Chlorella vulgaris induced DNA damage and apoptosis

OBJECTIVES:

The aim of this study was to determine the antiproliferative and apoptotic effects of hot water extracts of Chlorella vulgaris on hepatoma cell line HepG2.

INTRODUCTION:

The search for food and spices that can induce apoptosis in cancer cells has been a major study interest in the last decade. Chlorella vulgaris, a unicellular green algae, has been reported to have antioxidant and anti‐cancer properties. However, its chemopreventive effects in inhibiting the growth of cancer cells have not been studied in great detail.

METHODS:

HepG2 liver cancer cells and WRL68 normal liver cells were treated with various concentrations (0‐4 mg/ml) of hot water extract of C. vulgaris after 24 hours incubation. Apoptosis rate was evaluated by TUNEL assay while DNA damage was assessed by Comet assay. Apoptosis proteins were evaluated by Western blot analysis.

RESULTS:

Chlorella vulgaris decreased the number of viable HepG2 cells in a dose dependent manner (p < 0.05), with an IC₅₀ of 1.6 mg/ml. DNA damage as measured by Comet assay was increased in HepG2 cells at all concentrations of Chlorella vulgaris tested. Evaluation of apoptosis by TUNEL assay showed that Chlorella vulgaris induced a higher apoptotic rate (70%) in HepG2 cells compared to normal liver cells, WRL68 (15%). Western blot analysis showed increased expression of pro‐ apoptotic proteins P53, Bax and caspase‐3 in the HepG2 cells compared to normal liver cells WRL68, and decreased expression of the anti‐apoptotic protein Bcl‐2.

CONCLUSIONS:

Chlorella vulgaris may have anti‐cancer effects by inducing apoptosis signaling cascades via an increased expression of P53, Bax and caspase‐3 proteins and through a reduction of Bcl‐2 protein, which subsequently lead to increased DNA damage and apoptosis.     

Modulation of Ki67 and myogenic regulatory factor expression by tocotrienol-rich fraction ameliorates myogenic program of senescent human myoblasts

IntroductionReplicative senescence results in dysregulation of cell proliferation and differentiation, which plays a role in the regenerative defects observed during age-related muscle atrophy. Vitamin E is a well-known antioxidant, which potentially ameliorates a wide range of age-related manifestations. The aim of this study was to determine the effects of tocotrienol-rich fraction (TRF) in modulating the expression of proliferation- and differentiation-associated proteins in senescent human myoblasts during the differentiation phase.Material and methodsHuman skeletal muscle myoblasts were cultured until senescence. Young and senescent cells were treated with TRF for 24 h before and after differentiation induction, followed by evaluation of cellular morphology and efficiency of differentiation. Expression of cell proliferation marker Ki67 protein and myogenic regulatory factors MyoD and myogenin were determined.ResultsOur findings showed that treatment with TRF significantly improved the morphology of senescent myoblasts. Promotion of differentiation was observed in young and senescent myoblasts with TRF treatment as shown by the increased fusion index and larger size of myotubes. Increased Ki67 and myogenin expression with TRF treatment was also observed in senescent myoblasts, suggesting amelioration of the myogenic program by TRF during replicative senescence.ConclusionsTRF modulates the expression of regulatory factors related to proliferation and differentiation in senescent human myoblasts and could be beneficial for ameliorating the regenerative defects during aging.     

5-Azacytidine Is Insufficient For Cardiogenesis In Human Adipose-Derived Stem Cells

Background  

Adipose tissue is a source of multipotent adult stem cells and it has the ability to differentiate into several types of cell lineages such as neuron cells, osteogenic cells and adipogenic cells. Several reports have shown adipose-derived stem cells (ASCs) have the ability to undergo cardiomyogenesis. Studies have shown 5-azacytidine can successfully drive stem cells such as bone marrow derived stem cells to differentiate into cardiomyogenic cells. Therefore, in this study, we investigated the effect 5-azacytidine on the cardiogenic ability of ASCs.     

Tocotrienol rich fraction supplementation improved lipid profile and oxidative status in healthy older adults: A randomized controlled study

Background

Vitamin E supplements containing tocotrienols are now being recommended for optimum health but its effects are scarcely known. The objective was to determine the effects of Tocotrienol Rich Fraction (TRF) supplementation on lipid profile and oxidative status in healthy older individuals at a dose of 160 mg/day for 6 months.

Methods

Sixty-two subjects were recruited from two age groups: 35-49 years (n = 31) and above 50 years (n = 31), and randomly assigned to receive either TRF or placebo capsules for six months. Blood samples were obtained at 0, 3^rd and 6^th months.

Results

HDL-cholesterol in the TRF-supplemented group was elevated after 6 months (p < 0.01). Protein carbonyl contents were markedly decreased (p < 0.001), whereas AGE levels were lowered in the > 50 year-old group (p < 0.05). Plasma levels of total vitamin E particularly tocopherols were significantly increased in the TRF-supplemented group after 3 months (p < 0.01). Plasma total tocotrienols were only increased in the > 50 year-old group after receiving 6 months of TRF supplementation. Changes in enzyme activities were only observed in the > 50 year-old group. SOD activity was decreased after 3 (p < 0.05) and 6 (p < 0.05) months of TRF supplementation whereas CAT activity was decreased after 3 (p < 0.01) and 6 (p < 0.05) months in the placebo group. GPx activity was increased at 6 months for both treatment and placebo groups (p < 0.05).

Conclusion

The observed improvement of plasma cholesterol, AGE and antioxidant vitamin levels as well as the reduced protein damage may indicate a restoration of redox balance after TRF supplementation, particularly in individuals over 50 years of age.     

Antioxidant enzyme activity and malondialdehyde levels can be modulated by Piper betle,tocotrienol rich fraction and Chlorella vulgaris in aging C57BL/6 mice

OBJECTIVE:

The aim of this study was to determine the erythrocyte antioxidant enzyme activity and the superoxide dismutase, catalase, glutathione peroxidase, and plasma malondialdehyde levels in aging mice and to evaluate how these measures are modulated by potential antioxidants, including the tocotrienol-rich fraction, Piper betle, and Chlorella vulgaris.

METHOD:

One hundred and twenty male C57BL/6 inbred mice were divided into three age groups: young (6 months old), middle-aged (12 months old), and old (18 months old). Each age group consisted of two control groups (distilled water and olive oil) and three treatment groups: Piper betle (50 mg/kg body weight), tocotrienol-rich fraction (30 mg/kg), and Chlorella vulgaris (50 mg/kg). The duration of treatment for all three age groups was two months. Blood was withdrawn from the orbital sinus to determine the antioxidant enzyme activity and the malondialdehyde level.

RESULTS:

Piper betle increased the activities of catalase, glutathione peroxidase, and superoxide dismutase in the young, middle, and old age groups, respectively, when compared to control. The tocotrienol-rich fraction decreased the superoxide dismutase activity in the middle and the old age groups but had no effect on catalase or glutathione peroxidase activity for all age groups. Chlorella vulgaris had no effect on superoxide dismutase activity for all age groups but increased glutathione peroxidase and decreased catalase activity in the middle and the young age groups, respectively. Chlorella vulgaris reduced lipid peroxidation (malondialdehyde levels) in all age groups, but no significant changes were observed with the tocotrienol-rich fraction and the Piper betle treatments.

CONCLUSION:

We found equivocal age-related changes in erythrocyte antioxidant enzyme activity when mice were treated with Piper betle, the tocotrienol-rich fraction, and Chlorella vulgaris. However, Piper betle treatment showed increased antioxidant enzymes activity during aging.     

Impact of adipogenic differentiation on stemness and osteogenic gene expression in extensive culture of human adipose-derived stem cells

Introduction

Adipose tissue is a source of multipotent adult stem cells. Most studies on human adipose-derived stem cells (ASC) have been on the early passages. Studies in extensive expansion have not been well established yet. In this study, we aim to investigate the effects of extensive expansion on the adipogenic differentiation capability of ASC.

Material and methods

The ability of ASC to undergo adipogenic differentiation in extensive expansion was evaluated by morphological changes, differentiation assay by using Oil Red O staining and changes in the genes expression levels of adipogenic genes, osteogenic genes and stemness genes using quantitative polymerase chain reaction (qPCR) after induction.

Results

Morphological study showed that the formation of lipid droplets can be observed at all passages but decreased at P20 after induction. Data from qPCR showed that most adipogenicgenes expression increased significantlyat P5, P10 and P15 but decreased at P20 after induction. On the other hand, osteogenic genes showed no significant changes after adipogenic induction indicating low potentiality of adipogenic-induced ASC to become osteogenic cells. While stemness genes expression levels showed a decrease or no significant changes after adipogenic induction except Nanog3, which showed a significant increase at P15 and P20.

Conclusions

The ability of ASC to differentiate into mature adipogenic cells decreased after P10 and the decrease in the osteogenics gene expression level during adipogenic induction suggested that the osteogenesis and adipogenesis are not parallel events.