Therapeutic Potential of Tpl2 (Tumor Progression Locus 2) Inhibition on Diabetic Vasculopathy Through the Blockage of the Inflammasome Complex.

OBJECTIVE: Diabetic retinopathy, one of retinal vasculopathy, is characterized by retinal inflammation, vascular leakage, blood-retinal barrier breakdown, and neovascularization. However, the molecular mechanisms that contribute to diabetic retinopathy progression remain unclear. Approach and Results: Tpl2 (tumor progression locus 2) is a protein kinase implicated in inflammation and pathological vascular angiogenesis. Nε-carboxymethyllysine (CML) and inflammatory cytokines levels in human sera and in several diabetic murine models were detected by ELISA, whereas liquid chromatography-tandem mass spectrometry analysis was used for whole eye tissues. The CML and p-Tpl2 expressions on the human retinal pigment epithelium (RPE) cells were determined by immunofluorescence. Intravitreal injection of pharmacological inhibitor or NA (neutralizing antibody) was used in a diabetic rat model. Retinal leukostasis, optical coherence tomography, and H&E staining were used to observe pathological features. Sera of diabetic retinopathy patients had significantly increased CML levels that positively correlated with diabetic retinopathy severity and foveal thickness. CML and p-Tpl2 expressions also significantly increased in the RPE of both T1DM and T2DM diabetes animal models. Mechanistic studies on RPE revealed that CML-induced Tpl2 activation and NADPH oxidase, and inflammasome complex activation were all effectively attenuated by Tpl2 inhibition. Tpl2 inhibition by NA also effectively reduced inflammatory/angiogenic factors, retinal leukostasis in streptozotocin-induced diabetic rats, and RPE secretion of inflammatory cytokines. The attenuated release of angiogenic factors led to inhibited vascular abnormalities in the diabetic animal model. CONCLUSIONS: The inhibition of Tpl2 can block the inflammasome signaling pathway in RPE and has potential clinical and therapeutic implications in diabetes-associated retinal microvascular dysfunction.

Antihyperlipidemic activity of Allium chinense bulbs.

Allium chinense is a medicinal plant and nutritional food commonly used in Eastern Asia. In this study, we investigated the in vitro antioxidant activity (scavenging of α,α-diphenyl-ß-picrylhydrazyl free radical, total phenol content, reducing power, and total antioxidant activity) and constituents of various extracts from A. chinense. Moreover, we also studied the in vivo hypolipidemic effects of extracts on high-fat-diet Wistar rats. Ethanol extracts from A. chinense showed notable antioxidant activity, and its high-dose essential-oil extract both significantly reduced serum and hepatic total cholesterol, triglyceride, and low-density lipoprotein levels and increased serum high-density lipoprotein levels in high-fat-diet Wistar rats compared with those observed following treatment with the control drug probucol. Additionally, visceral fat in high-fat-diet Wistar rats was reduced. Furthermore, groups with high doses of essential-oil and residue extracts showed protective effects associated with histopathological liver alteration. These results suggested that A. chinense is a valuable plant worthy of further investigation as a potential dietary supplement or botanical drug.

Chemical compositions and physicochemical properties of the fiber-rich materials prepared from shoyu mash residue.

Fiber-rich materials including desalted shoyu mash residue (briefly referred as desalted mash residue, DMR), alcohol-insoluble solid (AIS), and water-insoluble solid (WIS) were prepared from shoyu mash residue, which is a filtration cake obtained during the isolation of shoyu by press filtration of fermented matrix in the final process. The DMR, AIS, and WIS contain rich dietary fiber of 52.4, 61.5, and 54.7 wt %, respectively. The DMR, AIS, and WIS all have significantly lower bulk densities, and higher water-holding capacities, oil-holding capacities, swelling abilities, and cation-exchange capacities than the control cellulose. These results indicated that the said fiber-rich materials prepared in this study all have the desired physicochemical properties for being used as satisfactory sources of dietary fibers or low-calorie bulk ingredients in food applications requiring oil and moisture retention. Furthermore, the said fiber-rich materials also have high contents of isoflavones, mainly daidzein and genistein, which are considered as the most bioavailable phytoestrogens, with a total amount of about 1200-1480 micromol/100 g (equal to daidzein of ca. 3040-3759 microg/g, or genistein of 3240-3996 microg/g). The results revealed that the said fiber-rich materials might be a potent fiber source for health foods.

Nε-carboxymethyllysine-mediated endoplasmic reticulum stress promotes endothelial cell injury through Nox4/MKP-3 interaction.

N(ε)-carboxymethyllysine (CML) is an important driver of diabetic vascular complications and endothelial cell dysfunction. However, how CML dictates specific cellular responses and the roles of protein tyrosine phosphatases and ERK phosphorylation remain unclear. We examined whether endoplasmic reticulum (ER) localization of MAPK phosphatase-3 (MKP-3) is critical in regulating ERK inactivation and promoting NADPH oxidase-4 (Nox4) activation in CML-induced endothelial cell injury. We demonstrated that serum CML levels were significantly increased in type 2 diabetes patients and diabetic animals. CML induced ER stress and apoptosis, reduced ERK activation, and increased MKP-3 protein activity in HUVECs and SVECs. MKP-3 siRNA transfection, but not that of MKP-1 or MKP-2, abolished the effects of CML on HUVECs. Nox4-mediated activation of MKP-3 regulated the switch to ERK dephosphorylation. CML also increased the integration of MKP-3 with ERK, which was blocked by silencing MKP-3. Exposure of antioxidants abolished CML-increased MKP-3 activity and protein expression. Furthermore, immunohistochemical staining of both MKP-3 and CML was increased, but phospho-ERK staining was decreased in the aortic endothelium of streptozotocin-induced and high-fat diet-induced diabetic mice. Our results indicate that an MKP-3 pathway might regulate ERK dephosphorylation through Nox4 during CML-triggered endothelial cell dysfunction/injury, suggesting that therapeutic strategies targeting the Nox4/MKP-3 interaction or MKP-3 activation may have clinical implications for diabetic vascular complications.

Magnesium status and association with diabetes in the Taiwanese elderly.

The average dietary intake of magnesium is below recommended dietary allowances in many affluent Western countries. Prolonged low magnesium intake tends to result in hypomagnesaemia which might increase the risk of chronic diseases in elderly people. A national population-based cross-sectional nutrition survey, the Elderly Nutrition and Health Survey in Taiwan (1999-2000), was used to investigate the magnesium status and association with diabetes in the Taiwanese elderly. Dietary magnesium intake was based on 24-hour dietary recalls. Blood biochemical parameters including plasma magnesium and blood glucose were also measured. Average magnesium intake was 250 mg in men and 216 mg in women, which is equivalent to 68-70% of relevant Taiwanese Dietary Reference Intakes. The mean plasma magnesium concentration was 0.903 mmol/L in men and 0.906 mmol/L in women. The prevalence of a plasma magnesium level of <0.7 mmol/L was 0.7-0.9% in the elderly, and that of <0.8 mmol/L was 8.0-9.1%. Elderly vegans had a significantly lower magnesium intake than ovo-lacto vegetarians and non-vegetarians. Diabetic men and women had significantly higher blood glucose levels than non-diabetics. The risk of diabetes was elevated 3.25 times at plasma magnesium levels<0.863 mmol/L. There was an inverse association between plasma magnesium concentration and the prevalence of diabetes. However, no association was found between diabetes and low dietary magnesium. Taiwanese elderly persons had suboptimal levels of dietary magnesium intake, which although may be sufficient to avoid overt magnesium deficiency, may not be sufficient to reduce the risk of diabetes in the elderly. Further prospective study is required to help explain the differential results between dietary and plasma magnesium levels.