Male Wistar rats were purchased from National Institute of Nutrition, Hyderabad, India and the study was strictly done under the guidelines approved by the animal ethical Committee of CSIR-Institute of Genomics and Integrative Biology, Delhi, India. They were maintained under standard conditions (12/12 hour light/dark cycle at 25±5ºC). They were divided into five groups having six rats per cage and are allowed to acclimatization for 1 week.
Group 1(G1) - Healthy control (chow diet); Group 2 (G2) - HFD 150mg/kg body weight and the methanolic extract of Dracaena cinnabari resin 300mg/kg body weight; Group 3 (G3) - HFD 150mg/kg body weight and standard drug atorvastatin 30mg/kg body weight; Group 4 (G4) - HFD 150mg/kg body …show more content…
After that the treatment was followed by oral administration of methanolic extract of Dracaena cinnabari (300mg/kg/day); aqueous extract of pure compound curcumin (300 mg/kg/day) and standard drug atorvastatin (30mg/kg/day) daily for one month. After thirty days administration of the respective drug, the experimental and control rats were sacrificed and blood was collected into a vacutainer containing ethylene diamine tetraacetic acid (EDTA).
Sample preparation for two-dimensional gel electrophoresis
The blood samples were centrifuged at 1,500 rpm for 15 min at 4°C; plasma was isolated and stored at -80°C. Sample preparation was carried out by removing salt from the plasma by centrifugation and concentrated using 3 kDa amicon columns (Merck Millipore, USA) .
Two-dimensional gel …show more content…
Briefly, gels were placed in fixative (50% methanol and 12% acetic acid) for 1 h, soaked in 50% ethanol for 30 min followed by 30% ethanol for 30 min. The gels were then sensitized for 1 min in 2% solution of sodium sulfite (Na2SO3) followed by rinsing three times with Mili-Q for 20 sec and were stained for 20 min in staining solution [1M Silver nitrate (AgNO3) and formaldehyde (HCHO)]. After staining, gels were rinsed thrice for 20 sec in distilled water and developed for 5–10 min in a developer [sodium carbonate (Na2CO3) and formaldehyde (HCHO)] and observed for the appearance of intense bands for 1–2 min. The reaction was stopped with 6% acetic acid solution and scanned using chemiDocTM MP imaging system (Bio-Rad,