Effect of chemical chaperones on glucose-induced lysozyme modifications.

Nonenzymatic glycation of biomacromolecules occurs due to the diabetes mellitus and ageing. A number of small molecules, known as chemical chaperones, stabilize protein conformation against thermal and chemically induced denaturation. These compounds are including: polyamines (e.g. spermine and spermidine), amino acids (e.g. lysine) and polyols (e.g. glycerol). In this study the effect of spermidine (Spd), spermine (Spm), and glycerol on glycation, structure and function of lysozyme (LZ), as an extra-cellular protein, by different techniques is investigated. LZ is incubated with or without glucose (50 or 100 mM) in the absence or presence of Spd/Spm/glycerol at 37 °C up to 16 weeks. All the observed changes of glycated-LZ in comparison with the native protein, including: increased fluorescence emission, alteration in the secondary and tertiary structure, and reduced electrophoretic mobility- indicate its structural changes that are accompanied with its reduced activity. Glucose in the presence or absence of Spd induces the protein dimerization, but glucose plus Spm induces its trimmerization. In contrast, glycerol inhibits the LZ glycation and prevents the large changes on its structure and function. Glucose binds lysine residues, decreases the protein positive charges and induces some alterations in its structure and activity. Polyamines also directly bind to LZ, increase its positive charges and hence induce more glycation; more conformational changes, oligomerization and its inactivation in the presence of glucose, but glycerol affect the protein environment and preserve protein from these harmful effects.

The effect of hot-tub therapy on serum Hsp70 level and its benefit on diabetic rats: a preliminary report.

PURPOSE: To carry out a preliminary study examining the efficacy of long-term hot-tub therapy (HTT) in the improvement of diabetic complications on streptozotocin-induced diabetic rats. MATERIALS AND METHODS: Male Wistar rats were immersed mid-sternum in a circulating water bath (42 degrees C for 30 min) to obtain a core body temperature of 41 degrees C; this process was repeated three times a week for 5 months. The blood was collected every month. Multiple parameters were examined for all rats including heat shock protein (Hsp70) level, serum glucose and insulin concentrations, advanced glycation end product (AGE) and glycated haemoglobin (HbA1c) formation, lipid profile and antioxidant defence system. Additionally, the chaperoning capacity of glycated Hsp70 was evaluated based on in vitro studies in which the refolding of denatured luciferase was compared to refolding by native Hsp70. RESULTS: HTT-treated diabetic rats showed a significant improvement in lipid profile, antioxidant capacity, insulin secretion and serum Hsp70 level and a significant decrease in AGE formation compared to the untreated diabetic rats. However, HTT had a borderline significant effect on weight and fasting blood glucose. Glycated Hsp70 lost its chaperoning ability to reactivate the denatured luciferase. CONCLUSION: A decrease in complications in diabetic rats after hot-tub therapy is shown here. An increase in the extracellular Hsp70 level due to HTT was observed. This increase may serve to protect the structure of proteins (e.g. preventing AGE formation), and the observed beneficial effects may be related to it.

Investigation of the mechanisms involved in the high-dose and long-term acetyl salicylic acid therapy of type I diabetic rats.

Diabetes mellitus has been classified as a conformational disease because of changes induced in the structure and function of proteins due to hyperglycemia. In this study, we investigated the effect of high-dose and long-term use of acetyl salicylic acid (ASA) on the streptozotocin-induced diabetic rats as a model of type I diabetes, with consideration on the structure and/or function of proteins. The N-[methylnitrosocarbamoyl]-d-glucosamine (streptozotocin)-induced diabetic rats together with the normal rats were studied for 5 months with and without receiving 100 mg/kg ASA in drinking water. All rats were investigated from different aspects such as heat shock protein (HSP) 70 level, serum glucose and insulin concentration, advanced glycated end product (AGE) and glycated hemoglobin (HbA1c) formation, lipid profile, high-density lipoprotein (HDL) functionality (paraoxonase1 and lecithin cholesterol acyltransferase activities), and the antioxidant system. In addition, the in vitro effect of ASA on the structure of albumin as a model protein was studied in the presence of glucose by spectroscopic techniques such as fluorometry and circular dichroism. The results show that ASA therapy causes a decrease in the glucose level and AGE and HbA1c formation, improves the lipid profile, HDL functionality, and the antioxidant capacity, induces serum HSP70, and overall decreases mortality of diabetic rats in comparison with the group without treatment. The conformation of glycated bovine serum albumin is different from the native form, and ASA retains the conformation of this protein similar to the native. The improving effect of ASA on diabetic rats is mostly due to its role as a chemopreventive agent in the structural conservation and protection of proteins involved in diabetes pathogenesis.