Masashi Inafuku

Coordinated Actions of Glyoxalase and Antioxidant Defense Systems in Conferring Abiotic Stress Tolerance in Plants

Being sessile organisms, plants are frequently exposed to various environmental stresses that cause several physiological disorders and even death. Oxidative stress is one of the common consequences of abiotic stress in plants, which is caused by excess generation of reactive oxygen species (ROS). Sometimes ROS production exceeds the capacity of antioxidant defense systems, which leads to oxidative stress. In line with ROS, plants also produce a high amount of methylglyoxal (MG), which is an α-oxoaldehyde compound, highly reactive, cytotoxic, and produced via different enzymatic and non-enzymatic reactions. This MG can impair cells or cell components and can even destroy DNA or cause mutation. Under stress conditions, MG concentration in plants can be increased 2- to 6-fold compared with normal conditions depending on the plant species. However, plants have a system developed to detoxify this MG consisting of two major enzymes: glyoxalase I (Gly I) and glyoxalase II (Gly II), and hence known as the glyoxalase system. Recently, a novel glyoxalase enzyme, named glyoxalase III (Gly III), has been detected in plants, providing a shorter pathway for MG detoxification, which is also a signpost in the research of abiotic stress tolerance. Glutathione (GSH) acts as a co-factor for this system. Therefore, this system not only detoxifies MG but also plays a role in maintaining GSH homeostasis and subsequent ROS detoxification. Upregulation of both Gly I and Gly II as well as their overexpression in plant species showed enhanced tolerance to various abiotic stresses including salinity, drought, metal toxicity, and extreme temperature. In the past few decades, a considerable amount of reports have indicated that both antioxidant defense and glyoxalase systems have strong interactions in conferring abiotic stress tolerance in plants through the detoxification of ROS and MG. In this review, we will focus on the mechanisms of these interactions and the coordinated action of these systems towards stress tolerance.

Peucedanum japonicum Thunb Inhibits High‐fat Diet Induced Obesity in Mice

This study examined the antiobese activity of Peucedanum japonicum Thunb (PJT) in mice. In the first experiment, 4‐week‐old C57BL/6 mice were fed seven different diets containing 15% corn oil and 0–20% PJT powder for 4 weeks. Feeding the 10% and 20% PJT diet suppressed the body weight gain and the accumulation of abdominal and subcutaneous fats. PJT reduced serum and liver levels of triglyceride and serum levels of leptin in a dose‐dependent manner. PJT intake decreased the proportion of saturated fatty acids and increased polyunsaturated and n‐3 fatty acids in the liver. To obtain more insight into the antiobese activity of PJT, its effect on lipid absorption and enzyme activities related to lipid metabolism was studied in the second experiment. There was an increased faecal excretion of triglyceride in mice fed 5% and 10% PJT diets. Fatty acid synthase activity was decreased while carnitine palmitoyltransferase activity was increased by 10% PJT intake. These findings pointed to the usefulness of PJT for the development of a safe natural agent to reduce obesity or body weight for the first time. The rationale for the lipid lowering mechanism of PJT and the candidate compound responsible for the observations have also been discussed. Copyright

Expression of terpenoid synthase mRNA and terpenoid content in salt stressed mangrove

The halophytic Kandelia candel and Bruguiera gymnorrhiza are ideal model for studying the molecular mechanisms of salinity tolerance in mangrove plants. The correlation between mRNA expression of four oxidosqualene cyclase (OSC) genes namely, KcMS multifunctional terpenoid synthase and KcCAS cyloartenol synthase (K. candel), BgbAS beta-amyrin synthase and BgLUS lupeol synthase (B. gymnorrhiza) and salt concentration was examined. mRNA level of KcMS was increased with salt concentration in both roots and leaves of K. candel. Similarly, salt stress increased the mRNA levels of BgLUS and BgbAS in the root of B. gymnorrhiza. This result suggests that the function of terpenoids in root is associated with the salt stress. In contrast to these observations, the mRNA level of KcCAS was not modulated by salt stress in the roots, and decreased in the leaves. These results therefore suggest that the terpenoids but not phytosterols play an important role to cope with the salt stress in mangrove root. The content and proportion of beta-amyrin and lupeol increased with salinity in the root of K. candel and B. gymnorrhiza, providing additional evidence for the protective role of terpenoids. However, beta-amyrin and lupeol in B. gymnorrhiza leaves decreased with salt concentration, suggesting that the physiological significance for the terpenoids in the leaf may differ from that for the root.

The tumor specific cytotoxicity of dihydronitidine from Toddalia asiatica Lam

Purpose: In recent years, a number of reports have shown the anticancer activity of plant extracts and phytoalkaloid. Methods: We have evaluated the cytotoxicity profiles of 157 extracts prepared from dietary or medical plants growing in the Okinawa island, using 10 different cell lines. In vitro cytotoxicity screening indicated the presence of a highly selective cytotoxic compound in the extract of Toddalia asiaticaLam. The known alkaloid (1,3)benzodioxolo(5,6-c)phenanthridine, 12,13-dihydro-2,3-dimethoxy-12-methyl-(dihydronitidine) was identified as an active material from this plant. This alkaloid had highly specific cytotoxicity to human lung adenocarcinoma (A549) cells. Results: The results of the fluorescence activated cell sorter (FACS) analysis and the measurement of caspase-3 activity showed that dihydronitidine induced specific apoptotic cell death in A549 cells. Gene expression analysis in the apoptotic cells found that dihydronitidine variously regulated the cell cycle related genes (CDK2 and CCNE), and up-regulated the cell death related genes specifically in tumor cells. Thus dihydronitidine manifested its characteristics in the tumor selective cytotoxicity, contrasting with the case of a known anticancer agent camptothecin (CPT). Microscopic observation further revealed the specific accumulation of dihydronitidine within the cytosolic organelle, but not in the nuclei of adenocarcinoma. No accumulation was observed with CPT in all cell lines. Conclusion: The data suggested that dihydronitidine toxicity targeted a particular intracellular organelle in the tumor cells.

Mechanism of arctigenin-mediated specific cytotoxicity against human lung adenocarcinoma cell lines

The lignan arctigenin (ARG) from the herb Arctium lappa L. possesses anti-cancer activity, however the mechanism of action of ARG has been found to vary among tissues and types of cancer cells. The current study aims to gain insight into the ARG mediated mechanism of action involved in inhibiting proliferation and inducing apoptosis in lung adenocarcinoma cells. This study also delineates the cancer cell specificity of ARG by comparison with its effects on various normal cell lines. ARG selectively arrested the proliferation of cancer cells at the G0/G1 phase through the down-regulation of NPAT protein expression. This down-regulation occurred via the suppression of either cyclin E/CDK2 or cyclin H/CDK7, while apoptosis was induced through the modulation of the Akt-1-related signaling pathway. Furthermore, a GSH synthase inhibitor specifically enhanced the cytotoxicity of ARG against cancer cells, suggesting that the intracellular GSH content was another factor influencing the susceptibility of cancer cells to ARG. These findings suggest that specific cytotoxicity of ARG against lung cancer cells was explained by its selective modulation of the expression of NPAT, which is involved in histone biosynthesis. The cytotoxicity of ARG appeared to be dependent on the intracellular GSH level.

Effect of Vaccinium ashei reade leaves on lipid metabolism in Otsuka Long-Evans Tokushima Fatty rats

The effects of blueberry leaf (BBL) on lipid metabolism were studied in obese rats. Feeding of BBL lowered levels of serum lipids and C-reactive protein and alleviated hepatic triglyceride accumulation in the rats. The hypolipidemic effect might be attributable to a reduction of lipogenesis and enhancement of lipolysis in the liver. These results suggest the use of blueberry leaf as a dietary hypolipidemic component.

Mukitake mushroom (Panellus serotinus) alleviates nonalcoholic fatty liver disease through the suppression of monocyte chemoattractant protein 1 production in db/db mice

Nonalcoholic fatty liver disease (NAFLD) is emerging as the most common liver disease in industrialized countries. Thus, the discovery of food components that would ameliorate NAFLD is of interest. Various mushrooms have been used in folk medicine for the treatment of lifestyle diseases in eastern countries and several compounds that modulate immune system, lower blood lipid levels, inhibit tumor and viral action have been isolated from them. In this study, we tested whether feeding Panellus serotinus (Mukitake) to db/db mice protects them from hepatic injury. After 4 weeks of feeding, hepatomegaly, hepatic triglyceride accumulation and elevated hepatic injury markers in serum were markedly alleviated in Mukitake-fed db/db mice compared with control mice. These effects were partly attributable to the enhancement of lipolytic enzyme activity and the suppression of lipogenic enzyme activities due to the Mukitake diet. The severe hyperinsulinemia in control db/db mice tended to attenuate in Mukitake-fed mice due to an enhanced production of adiponectin, which improves insulin sensitivity. Moreover, production of monocyte chemoattractant protein 1 (MCP1), an inflammatory cytokine, was markedly suppressed by the Mukitake diet. In addition, water-soluble extracts of Mukitake powder showed an inhibitory effect on inhibitor of kappaB (IkappaB) kinase (IKK) beta, whose activation is required for nuclear factor kappaB (NFkappaB)-mediated inflammatory response. We speculate that the development and progression of NAFLD was prevented by the reduction of MCP1 production through the interference in the IKKbeta-NFkappaB signaling pathway in Mukitake-fed db/db mice.

Partially purified Peucedanum japonicum Thunb extracts exert anti-obesity effects in vitro.

OBJECTIVE Crude ethanolic extract of Peucedanum japonicum Thunb (PJT) has been identified as an antiobesity and antidiabetic candidate. The aim of this study was to assess the mechanisms related to the antiobesity activity of partially purified PJT ethanol extract in vitro. METHODS PJT ethanol extract (EE) was partitioned into hexane phase (HP) and residual water phase. The effect of PJT extracts on triacylglycerol (TG) content, lipid metabolism-related gene expression, and energy expenditure was assessed in vitro in 3T3-L1, HepG2, and C2C12 cells. Furthermore, the active components in PJT extracts were partially profiled by high-performance liquid chromatography (HPLC) analysis. RESULTS The HP significantly down-regulated lipogenic gene expressions in hepatocytes, inhibited TG accumulation, and decreased the size of 3T3-L1 adipocytes. Moreover, the inhibition of lipid accumulation was at maximum during the adipocyte maturation stage. Furthermore, we found an increase in fat hydrolysis by HP in 3T3-L1. In C2C12 myotubes, the HP tended to enhance energy expenditure. HPLC analysis demonstrated that hydrophobic compounds available in the HP were responsible for antiobesity, which corresponded to the latter peaks on the HPLC chromatogram of EE. CONCLUSION Our study demonstrated that the HP plays a crucial role in regulating lipid metabolism-related gene expressions and energy expenditure in vitro. The results thus provide insight on the activity of HP involved in suppressing obesity and its chemical entities.

Effect of Vaccinium ashei reade Leaf Extracts on Lipid Metabolism in Obese OLETF Rats

The effects of a hot water extract and fractional extracts from rabbiteye blueberry (Vaccinium ashei reade) leaves (BBL) on lipid metabolism were studied in obese Otsuka Long-Evans Tokushima Fatty (OLETF) rats. Feeding the hot water extract and fractional extracts from BBL alleviated hepatic triglyceride accumulation in the rats. Additionally, feeding with the flavonol glycoside (FG) and proanthocyanidin (PA) fractions lowered serum cholesterol levels in the obese rats. The results from measurements of the hepatic enzyme activity indicate that the hypolipidemic effects of the hot water extract and the PA fraction might be attributable to enhanced lipolysis in the liver. The reduced serum levels of C-reactive protein, an inflammatory cytokine, by the chlorogenic acid + rutin fraction and FG fraction might be associated with alleviating the metabolic abnormalities in obese rats. These results indicate that the BBL extracts, and especially FG and PA, exerted hypolipidemic effects on obese OLETF rats and suggest that an infusion of BBL can be useful as a dietary hypolipidemic component.

Antiatherosclerotic function of Kokuto, Okinawan noncentrifugal cane sugar.

In the present study, we investigated the effect of phenolic compounds (PCs) and policosanol of Kokuto, Okinawan noncentrifugal cane sugar, on the development of atherosclerosis. A total of 67 male Japanese quail were divided into eight dietary groups in trial 1. The dietary groups were fed the atherosclerotic diet (AD) containing 5% corn oil, 2% cholesterol, and 30% sucrose or seven different types of Kokuto. Dietary intakes of Kokuto notably prevented the development of atherosclerosis. Furthermore, there was a significant negative correlation between the serum radical scavenging activity and the degree of atherosclerosis in the dietary groups. In trial 2, a total of 63 Japanese quail were fed AD with sucrose, Kokuto, PC extracts from Kokuto, wax extracts from sugar cane, octacosanol, vitamin C, and vitamin E. As a result, the supplementation of the diet with Kokuto and PCs significantly reduced the development of atherosclerosis as compared with the ingestion of AD with sucrose. In conclusion, these findings suggest that, among various components of Kokuto, PCs play a central role for the prevention of experimental atherosclerosis in Japanese quail.