Evaluation of chronic toxicological profile of herbo-mineral formulations: Shwaskas Chintamani Rasa and its marketed formulation namely Kas Shwas Hari Rasa.

BACKGROUND: Shwaskas Chintamani Rasa (SKC) and Kas Shwas Hari Rasa (KSH) are the Ayurvedic herbo-mineral formulations. These Ayurvedic formulations contain heavy metals which is the reason of concern and might bring up the safety issue. OBJECTIVE: This research article is aimed to study chronic toxicity of SKC and KSH for safety aspect in Wistar rats. MATERIAL AND METHOD: A study group of 220 healthy rats were divided into six groups. These rats were administered with SKC and KSH formulations where both the formulations were administered for 180 consecutive days. SKC was administered at doses of 58 mg/kg (equivalent to therapeutic dose i.e. TD), 145 mg/kg (2.5 TD), 290 mg/kg (5 TD) and KSH was administered at dose of 58 mg/kg (TD). According to OECD guideline 452, the effect of these formulations was examined on hematology, serum biochemistry and histopathology of various organs. RESULTS: Both the formulations did not produce any signs or symptoms of treatment related toxicity in both male and female Wistar rats at therapeutic dose (TD), 2.5 times TD and 5 times TD. CONCLUSION: Based on these findings, the NOAEL (No observed adverse effect level) for test formulations SKC and KSH tablets in male and female wistar rats concluded to be preclinically safe.

Assessment of Chronic Toxicity of an Ayurvedic Herbo-Metallic Formulation Rasaraj Rasa in Wistar Rats.

Objectives: This study aimed to assess the adverse effects of Rasaraj Rasa tablets after repeated oral administration for 180 days in Wistar rats. Methods: Wistar rats were divided into five groups, of which three were treated with 54, 162, and 270 mg/kg body weight of Rasaraj Rasa, respectively, which correspond to one, three, and five times the proposed human therapeutic dose, for 180 days consecutively. The fifth group (satellite) also received 270 mg/kg body weight of Rasaraj Rasa for 180 days. Body weight and food intake were measured weekly. At the end of the study, all rats were sacrificed, and their blood, serum, and organs were collected and examined using hematology, serum biochemistry, gross pathology, and histopathology tests. In contrast, the satellite group was kept for 4 weeks after treatment. Results: No significant treatment-related toxicological findings were observed in the clinical features, body weight, laboratory findings, and pathological findings of the high-dose treated groups, when compared to those of the control group. Conclusion: The no-observed-adverse-effect-level for Rasaraj Rasa in Wistar rats is set at 270 mg/kg body weight.

Acceleration of protein glycation by oxidative stress and comparative role of antioxidant and protein glycation inhibitor.

Hyperglycemia in diabetes causes protein glycation that leads to oxidative stress, release of cytokines, and establishment of secondary complications such as neuropathy, retinopathy, and nephropathy. Several other metabolic disorders, stress, and inflammation generate free radicals and oxidative stress. It is essential to study whether oxidative stress independently enhances protein glycation leading to rapid establishment of secondary complications. Oxidative stress was experimentally induced using rotenone and Fenton reagent for in vivo and in vitro studies, respectively. Results showed significant increase in the rate of modification of BSA in the form of fructosamine and protein-bound carbonyls in the presence of fenton reagent. Circular dichroism studies revealed gross structural changes in the reduction of alpha helix structure and decreased protein surface charge was confirmed by zeta potential studies. Use of rotenone demonstrated enhanced AGE formation, ROS generation, and liver and kidney tissue glycation through fluorescence measurement. Similar findings were also observed in cell culture studies. Use of aminoguanidine, a protein glycation inhibitor, demonstrated reduction in these changes; however, a combination of aminoguanidine along with vitamin E demonstrated better amelioration. Thus, oxidative stress accelerates the process of protein glycation causing gross structural changes and tissue glycation in insulin-independent tissues. Use of antioxidants and protein glycation inhibitors in combination are more effective in preventing such changes and could be an effective therapeutic option for preventing establishment of secondary complications of diabetes.

Early metabolic changes in the gut leads to higher expression of intestinal alpha glucosidase and thereby causes enhanced postprandial spikes.

AIMS: Prediabetes manifests several years earlier, before it progresses to diabetes. It is essential to track the earliest metabolic changes occurring in the prediabetic state and to understand the precise mechanism of how diabetes is initiated. MAIN METHODS: Alpha glucosidase was isolated from rat intestine and assayed using maltose as substrate. In vitro glycation of the enzyme was studied using varying fructose content through measurement of fructosamine, general and specific fluorescence. In vivo experiments were carried out through feed of 4 g fructose per day. Protein expression was studied using western blot and mRNA expression using RT-PCR method. KEY FINDINGS: Fructose inhibits alpha glucosidase to the extent of 48.97% in 4 h at 2.5 M concentration. In vivo studies demonstrated an inhibition of 56.96% in three days. Activity was found to rise by seven days and normalized by 10 days. Protein expression was found to increase by 10.56 fold and SI mRNA by 41.84 fold on 10 days of fructose feed. Long term fructose feed for 60 days demonstrated increase in alpha glucosidase activity by 2.12 fold and increase in postprandial glucose spike. SIGNIFICANCE: Glycation of alpha glucosidase causes inhibition of the enzyme activity leading to compensation through higher protein expression. Long term fructose feed leads to more than two fold increase in enzyme activity causing postprandial spikes and ultimately manifesting as diabetes mellitus.