Huachen Wei

Antioxidant and Antipromotional Effects of the Soybean Isoflavone Genistein

Abstract Antioxidant and antipromotional effects of the soybean isoflavone genistein have been studied in HL-60 cells and the mouse skin tumorigenesis model. Effects of structure-related flavone/isoflavones on hydrogen peroxide (H2O2) production by 12-O-tetradecanoylphorbol-13-acetate (TPA)-activated HL-60 cells and superoxide anion (O2 −) generation by xanthine/xanthine oxidase were compared. Of tested isoflavones, genistein is the most potent inhibitor among TPA-induced H2O2 formation by (dimethyl sulfoxide) DMSO-differentiated HL-60 cells, daidzein is second, and aplgenin and biochanin A show little effect. In contrast, genistein, aplgenin, and prunectin are equally potent in inhibiting O2 − generation by xanthine/xanthine oxidase, with daidzein showing a moderate inhibitory effect and biochanin A exhibiting no effect. These results suggest that the antioxidant properties of isoflavones are structurally related and the hydroxy group at Position 4′ is crucial in both systems. Dietary administration of 250 ppm genistein for 30 days significantly enhances the activities of antioxidant enzymes in the skin and small intestine of mice. Further studies show that genistein significantly inhibits TPA-induced proto-oncogene expression (c-fos) in mouse skin in a dose-dependent manner. In a two-stage skin carcinogenesis study, low levels of genistein (1 and 5 μmol) significantly prolong tumor latency and decrease tumor multiplicity by approximately 50%. We conclude that genisteins antioxidant properties and antiproliferative effects may be responsible for its anticarcinogenic effect. Its high content in soybeans and relatively high bioavailability favor genistein as a promising candidate for the prevention of human cancers.

Inhibition of tumor Promoter‐induced hydrogen peroxide formation in vitro and in vivo by genistein

Here we report that genistein, a soybean isoflavone, strongly inhibits tumor promoter-induced H2O2 formation both in vivo and in vitro. Genistein suppressed H2O2 production by 12-O-tetradecanoylphorbol-13-acetate- (TPA) stimulated human polymorphonuclear leukocytes (PMNs) and HL-60 cells in a dose-dependent manner over the concentration range 1-150 microM. Human PMNs were more sensitive to the inhibitory effect of genistein than HL-60 cells (50% inhibitory concentration 14.8 and 30.2 microM, respectively). In addition, genistein moderately inhibited superoxide anion formation by HL-60 cells and scavenged exogenously added H2O2 under the same conditions as in cell culture. However, the H2O2-scavenging effect of genistein was about 50% lower than its inhibition of cell-derived H2O2 formation at all concentrations. In the CD-1 mouse skin model, genistein strongly inhibited TPA-induced oxidant formation, edema, and PMN infiltration in mouse skin. Inhibition of TPA-mediated H2O2 in vivo may result from decreased cell-derived H2O2 formation, scavenging of H2O2 produced, and/or suppression of PMN infiltration into the dermis. The antioxidant properties of genistein may be responsible for its anticarcinogenic effects, and the dietary availability of genistein makes it a promising candidate for the prevention of human cancers.

ALTERATIONS OF ANTIOXIDANT ENZYMES AND OXIDATIVE DAMAGE TO MACROMOLECULES IN DIFFERENT ORGANS OF RATS DURING AGING

Oxygen free radicals have been hypothesized to play an important role in the aging process. To investigate the correlation between the oxidative stress and aging, we have determined the levels of oxidative protein damage and lipid peroxidation in the brain and liver, and activities of antioxidant enzymes in the brain, liver, heart, kidney, and serum from the Fisher 344 rats at ages of 1, 6, 12, 18, and 24 months. The results showed that the level of oxidative protein damage (measured as carbonyl content) in the brain and liver was significantly higher in older animals than in young animals. No statistical difference was observed in the lipid peroxidation of the liver and brain between young and old animals. The activities of antioxidant enzymes in most tissues displayed an age-dependent decline. Superoxide dismutases in the heart, kidney, and serum, glutathione peroxidase activities in the serum and kidney, and catalase activities in the brain, liver, and kidney, significantly decreased during aging. Cytochrome c oxidase, an enzyme involved in electron transport in mitochondria, initially increased, but subsequently decreased in the aged brain, whereas no significant alteration was observed in the liver mitochondrial antioxidant enzymes. The present studies suggest that the accumulation of oxidized proteins during aging is most likely to be linked with an age-related decline of antioxidant enzyme activities, whereas lipid peroxidation is less sensitive to predict the aging process.

Effect of dietary genistein on antioxidant enzyme activities in SENCAR mice

Dietary administration of the soybean isoflavone genistein (50 and 250 ppm) for 30 days significantly increases the activities of antioxidant enzymes in various organs of SENCAR mice. Feeding a 250-ppm genistein diet to SENCAR mice significantly increases the activities of catalase in small intestine, liver, and kidney, the activities of superoxide dismutase and glutathione peroxidase in skin, and the activity of glutathione reductase in skin and small intestine. Feeding 50 ppm genistein to SENCAR mice results in elevated catalase activity in the small intestine and increases glutathione-S-transferase activities in skin, small intestine, liver, kidney, and lung. Dietary genisteins greatest enhancement of antioxidant enzyme activities occurred in skin and small intestine. Our results suggest that dietary genistein enhances the activities of antioxidant enzymes in various organs, which may be a mechanism(s) of genisteins chemopreventive action.

Isoflavone Genistein: Photoprotection and Clinical Implications in Dermatology

Genistein is a soybean isoflavone with diverse biological activities. It is a potent antioxidant, a specific inhibitor of protein tyrosine kinase, and a phytoestrogen. In recent years, increasing evidence has accumulated that this natural ingredient shows preventative and therapeutic effects for breast and prostate cancers, postmenopausal syndrome, osteoporosis, and cardiovascular diseases in animals and humans. In the past decade we have conducted a series of studies and demonstrated that genistein has significant antiphotocarcinogenic and antiphotoaging effects. Genistein substantially inhibits skin carcinogenesis and cutaneous aging induced by ultraviolet (UV) light in mice, and photodamage in humans. The mechanisms of action involve protection of oxidative and photodynamically damaged DNA, downregulation of UVB-activated signal transduction cascades, and antioxidant activities. In this article, we review the biological activities of genistein, as well as published and unpublished research from our laboratory. In addition, we discuss the potential application of genistein to clinical dermatology.

Genistein Alters the Ontogeny of Mammary Gland Development and Protects Against Chemically-Induced Mammary Cancer in Rats

Abstract Breast cancer is the most common cancer in US females and is the second leading cause of cancer death among women. By contrast, Asian women consuming a traditional diet high in soy products have a relatively low incidence of breast cancer. Asians who emigrate to the United States and adopt a Western diet lose this protection. Soy-based diets are high in phytoestrogens, and one of these components is genistein. Using the dimethylbenz(a)anthracene (DMBA) mammary cancer rodent model, we have investigated the breast cancer protective potential of genistein. Our results demonstrate that neonatal and prepubertal genistein treatments altered the ontogeny of the mammary gland and rendered the adult animals less susceptible to chemically-induced mammary cancer. Neonatal genistein treatment did not significantly alter the rate of formation and persistence of DMBA-DNA adducts in the mammary gland. While high concentrations of genistein during the neonatal period caused adverse effects on ovarian follicular development, prepubertal genistein treatment did not appear to be toxic in either the female reproductive tract or the endocrine system.

Shikonin modulates cell proliferation by inhibiting epidermal growth factor receptor signaling in human epidermoid carcinoma cells

Shikonin isolated from the roots of the Chinese herb Lithospermum erythrorhizon has been associated with anti-inflammatory properties. We evaluated shikonins chemotherapeutic potential and investigated its possible mechanism of action in a human cutaneous neoplasm in tissue culture. Shikonin preferentially inhibits the growth of human epidermoid carcinoma cells concentration- and time-dependently compared to SV-40 transfected keratinocytes, demonstrating its anti-proliferative effects against this cancer cell line. Additionally, shikonin decreased phosphorylated levels of EGFR, ERK1/2 and protein tyrosine kinases, while increasing phosphorylated JNK1/2 levels. Overall, shikonin treatment was associated with increased intracellular levels of phosphorylated apoptosis-related proteins, and decreased levels of proteins associated with proliferation in human epidermoid carcinoma cells.

Protective effects of lycopene against ultraviolet B-induced photodamage.

Lycopene, an acyclic hydrocarbon carotenoid found in tomatoes and their products, is a well-established potent antioxidant, and its anticancer properties have been shown in cultured cells and animal models. We investigated the protective effects of two concentrations of topical lycopene against acute ultraviolet B (UVB)-induced photodamage. Application of lycopene dose dependently inhibited UVB-induced ornithine decarboxylase (P < 0.05) and myeloperoxidase (P < 0.05) and significantly reduced bifold skin thickness (P < 0.05). Immunohistochemical staining revealed increased active caspase-3 of apoptotic pathway in the UVB-exposed group compared with the unexposed control. Application of topical lycopene prevented the cleavage of caspase-3. UVB irradiation completely diminished proliferating cell nuclear antigen (PCNA), and the untreated skin maintained positively stained cells throughout the basal epidermis. Topical application of lycopene significantly reversed UVB-induced PCNA inhibition, and normal PCNA staining was restored in the lycopene-treated skin. Our results suggest that topical lycopene is able to exert its protective effects against acute UVB-induced photodamage. Furthermore, it may act as a preventative agent via inhibition of epidermal ornithine decarboxylase activity, reducing inflammatory responses, maintaining normal cell proliferation, and possibly preventing DNA damage as indicated by blocking the necessitating step of apoptosis following UVB injury.

The preventive and therapeutic potential of the squalene-containing compound, Roidex, on tumor promotion and regression

Recent scientific evidence has shown free radicals or reactive oxygen species (ROS) to play an important role in the initiation and progression of cancer. Many radical scavengers have also been found to help reduce the attacks by these ROS. Interestingly, the ROS scavengers that have been investigated are naturally occurring compounds such as vitamins C and E. Roidex is a formulation of squalene, vitamin e, and aloe vera. It was our goal to investigate whether Roidex was able to prevent the development of chemically induced cancer and to cause regression of any tumors already formed in a mouse skin model. In the prevention study, skin tumors were initiated in 50 female CD-1 mice with 7,12-dimethylbenz[a]-anthracene (DMBA) and promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA). The mice were treated with either mineral oil, 5% squalene, or Roidex. At the end of the prevention study, there was a 33.34% incidence to tumors (multiplicity of 1.40) in the mineral oil-treatment group, 26.67% (multiplicity of 0.467) in the 5% squalene and Roidex groups, respectively. The tumor regression study involved the selection of mice with tumors and possible regression of these tumors with Roidex treatment. There was a regression of 33.34% of the tumors in the Roidex-treated group (39 tumors to 26 tumors) compared to the non-treated group whose tumors regressed only 3.44% (29 tumors to 28 tumors).

Effects of caloric restriction on age-related oxidative modifications of macromolecules and lymphocyte proliferation in rats.

Decreased immune function associated with aging has been demonstrated in both humans and animals. We hypothesize that reactive oxygen species (ROS)-mediated damage to biological macromolecules may contribute to compromised immune response during aging. In this study, we compared the levels of lipid peroxidation and oxidatively modified proteins in plasma and splenocytes, and the mitogen-induced T lymphocyte proliferation in ad lib-fed (AL) and caloric restricted (CR) Fischer 344 x BNF1 male rats at the ages of 5, 18, and 31 months. The results show that AL rats exhibit an age-related decrease in proliferative response of splenic lymphocytes to phytohemagglutinin (PHA) and concanavalin A (Con A). This functional decline in T-lymphocytes during aging is inversely correlated to the levels of both lipid peroxidation and protein carbonyl in the plasma and splenic lymphocytes. Caloric restriction, however, can partially reverse the age-dependent decrease in T lymphocyte proliferation and significantly reduce lipid peroxidation and protein carbonyl contents in plasma and splenocytes. The above observations support the hypothesis that the age-associated declines in immune function are related to the oxidative modification of biological macromolecules, which in turn may lead to enzyme inactivation, membrane disruption, and cell senescence. One of the mechanisms by which caloric restriction reverses declined immune function in aged rats is hypothesized to be through reduction in ROS production and thereby protection of cellular macromolecules against oxidative damage.