Development of high 1-deoxynojirimycin (DNJ) content mulberry tea and use of response surface methodology to optimize tea-making conditions for highest DNJ extraction

Mulberry 1-deoxynojirimycin (DNJ), a potent α-glucosidase inhibitor, suppresses postprandial blood glucose, thereby possibly preventing diabetes mellitus. At present, mulberry dry teas are commercially supplied as functional foods in many countries, but these products may not provide an effective dose (6 mg DNJ/60 kg human wt) due to their low DNJ content (about 100 mg/100 g of dry wt). Therefore, development of tea with higher DNJ content is desirable. To do this, we investigated distribution of DNJ content and α-glucosidase inhibitory activity in 35 Thai mulberry varieties. DNJ content in young leaves varied among mulberry varieties from 30 to 170 mg/100 g of dry leaves. Varieties having highest DNJ content were Kam, Burirum 60 and Burirum 51. Leaf position affected DNJ content: shoots > young leaves > mature leaves. DNJ concentration and α-glucosidase inhibitory activity were highly correlated (r = 0.84), suggesting that α-glucosidase inhibitory activity of mulberry leaves is mainly due to DNJ. Consequently, high DNJ content mulberry tea was produced from shoots of varieties such as Burirum 60, which contains 300 mg/100 g of dry wt. Tea-making conditions were optimized for highest DNJ extraction using response surface methodology. Approximate 95% of total DNJ in high DNJ content dry tea was extracted when temperature was maintained at 98 °C for 400 s; these conditions could be applicable for preparation of commercial products with high DNJ content. One cup (230 ml, a normal serving) of DNJ-enriched mulberry tea contained enough DNJ (6.5 mg) to effectively suppress postprandial blood glucose.     

Amadori-Glycated Phosphatidylethanolamine,a Potential Marker for Hyperglycemia,in Streptozotocin-Induced Diabetic Rats

It has been demonstrated in vivo that lipid glycation products such as Amadori-glycated phosphatidylethanolamine (Amadori-PE) accumulate in the plasma of diabetic humans and animals, but how lipid glycation products are formed under hyperglycemic conditions are not clear. We sought to clarify the occurrence of lipid glycation and its relationships with lipid peroxidation and protein glycation during the development of hyperglycemia using the streptozotocin (STZ)-induced diabetic rat model. A significant increase in Amadori-PE was observed in STZ rats 7 days after STZ treatment, and Amadori-PE (especially 18:0–20:4 Amadori-PE) was found at high levels in the blood and in organs that are strongly affected by diabetes, such as the kidney. Significant changes in Amadori-PE appeared to occur prior to changes in levels of oxidized lipids, which increased after 21–28 days. In addition, accumulation of Nε-(carboxymethyl)lysine (CML), a protein glycation product, proceeded somewhat more slowly and moderately than that of Amadori-PE, suggesting that Amadori-PE and CML are early and advanced glycation products, respectively. Our results suggest that Amadori-PE may be a useful predictive marker for hyperglycemia, particularly in the early stages of diabetes. Similar speculations have been made from previous human studies, but this study provides a direct evidence to support the speculations in rat study.     

A novel gelatin crosslinking method retards release of mulberry 1-deoxynojirimycin providing a prolonged hypoglycaemic effect

Mulberry 1-deoxynojirimycin (DNJ), a potent α-glycosidase inhibitor, has therapeutic potency in the suppression of postprandial blood glucose levels thereby possibly preventing diabetes mellitus. However, DNJ has a relatively short half-life in vivo (about 2 h). Therefore, several doses of mulberry DNJ are required to achieve optimal therapeutic results. This study aimed to delay the release of mulberry DNJ with biodegradable matrices to maintain the intestinal DNJ concentration and prolong the hypoglycaemic effect in vivo. A novel, simple, and commercially viable method was adopted to develop DNJ-entrapped microspheres (DNJ-MSs). A higher extent of crosslinking and the larger sized DNJ-MS decreased the rate of mulberry DNJ release in vitro. Consequently, an in vivo study was performed in Wistar rats over a 6 h period. The area under curve (AUC) of rats with DNJ-MS was significantly increased, compared to animals dosed with mulberry powder (control). DNJ-MS suppressed postprandial glucose from sucrose administration at the initial and 3 h time points indicating a prolonged hypoglycaemic effect.     

γ-Tocotrienol Reduces Squalene Hydroperoxide-Induced Inflammatory Responses in HaCaT Keratinocytes

Squalene hydroperoxide (SQ-OOH), the primary peroxidation product of squalene (SQ), accumulates at the surface of sunlight-exposed human skin. There are however only a few studies on the pathogenic actions (i.e., inflammatory stimuli) of SQ-OOH. Here, we evaluated whether SQ-OOH induced inflammatory responses in immortalized human keratinocytes (HaCaT). We found that SQ-OOH caused an increase in the expression of inflammatory genes such as the interleukins as well as cyclooxygenase-2 (COX-2). In concordance with the upregulation of COX-2 mRNA, SQ-OOH enhanced reactive oxygen species generation, nuclear factor kappa B activation, COX-2 protein expression, and prostaglandin E2 production. Therefore, the pro-inflammatory effects of SQ-OOH may be mediated in part via COX-2. On the other hand, γ-tocotrienol (γ-T3, an unsaturated form of vitamin E) was found to ameliorate the SQ-OOH actions. These results suggest that SQ-OOH induces inflammatory responses in HaCaT, implying that SQ-OOH plays an important role in inflammatory skin disorders. As a preventive strategy, inflammation could be reduced via the use of γ-T3.