EVALUATION OF RENAL EPITHELIAL CELL PROTEIN UNDER STRESS CONDITION
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2016, Vol 8, Issue 11
Abstract
Objective: Proteins are an important component of cells which are involved in various cellular functions. Different kind of stressing conditions has different responses in the components of the protein synthesis system. Super saturation condition in kidney environment leads to crystallization process. Crystals thus formed injure the surrounding cells and result in reactive oxygen species (ROS) formation. There might be some changes in the protein synthesis when the kidney cells enter in oxidative stress. In the present study, kidney cell lines were exposed to oxidative stress and their proteins were analyzed using Bradford analysis and SDS-PAGE.Methods: Vero cells were obtained from NCCS Pune and cultured in DMEM (Dulbecco’s Modified Eagle’s Medium) and maintained in a humidified incubator at 37 °C with 5% CO2. Calcium phosphate (CaP) crystals were prepared by the homogeneous system. After FTIR analysis crystals were used to injure Vero cell line. H2O2 was also used to injure the Vero cells. Intracellular protein was extracted from healthy cells and injured cells (with CaP crystals and H2O2). Ammonium sulfate precipitation method was used for the isolation of extracellular protein from the media of healthy and injured cells. Bradford method was used for the quantitative estimation of protein. Extracted proteins were analyzed by SDS-PAGE.Results: Amount of intracellular and extracellular protein of normal cells was 4.84±0.004µg/ml. Intracellular protein of CaP injured and H2O2 injured cells were 10.59±0.003 µg/ml and 10.78±0.011µg/ml respectively. While extracellular protein of injured cells was nearly 4 µg/ml. Intracellular protein bands ranging from 14.3 to 97.4 kDa was observed in healthy cells. Protein bands of ~40kDa and ~20kDa was absent in H2O2 and CaP injured intracellular protein extract. Two extracellular protein bands of 66kDa and ~60kDa were present in injured cells and healthy cells. Conclusion: When exposed to oxidative stress several proteins are oxidized decreasing the activity of many metabolic pathways. In the present study amount of intracellular protein increases when cells are injured with CaP or H2O2. While extracellular protein remains more or less same in both healthy and injured condition of cells. In SDS-PAGE analysis few bands were missing in the intracellular extract of injured cells. These results indicate that the amount of protein varies when cells are injured with CaP and H2O2.
Authors and Affiliations
Priyadarshini . , Kanika Jain, Rajeev Sood
Keywords
Related Articles
- EP ID EP575931
- DOI 10.22159/ijpps.2016v8i11.14310
- Views 67
- Downloads 0
MOLECULAR BEACON PROBE BASED PROMOTER MOTIFS VALIDATION IN ANOXIA RESPONSIVE DIFFERENTIALLY EXPRESSED GENES AND THEIR IN SILICO INTERACTION STUDIES WITH AP2/EREBP TF IN RICE (ORYZA SATIVA L.)
Objective: Progressive evolution in molecular biology revealed the differential expression of genes and their regulatory mechanism in rice under anoxia. In addition to that the consensus promoter motifs (GCC and TCC box)...
SYNTHESIS, ANTIMICROBIAL ACTIVITY AND MOLECULAR DOCKING STUDY OF SOME NEW N-BENZYL AND N-BENZOYL-3-INDOLYL HETEROCYCLES
Objective: Chalcones are one of the major classes of the natural products, which display a wide range of pharmacological properties. Also, chalcones are well-known intermediates for synthesizing various heterocyclic comp...
PREPARATION AND EVALUATION OF METRONIDAZOLE SUSTAINED RELEASE FLOATING TABLETS
Objective: The objective of the present study is the preparation of metronidazole (MZ) floating tablets that are designed to retain in the stomach for a long time for better eradication of Helicobacter Pylori (H. pylori)...
UV SPECTROPHOTOMETRIC METHOD FOR SIMULTANEOUS ESTIMATION OF TRAMADOL HYDROCHLORIDE AND ACECLOFENAC IN BULK AND TABLET DOSAGE FORM
Objective: To develop a simple, economic and validated UV spectrophotometric method for the simultaneous estimation of aceclofenac and tramadol hydrochloride in bulk and tablet dosage form.Methods: The UV spectrophotomet...
STRUCTURAL AND FUNCTIONAL ANALYSIS OF AF9-MLL ONCOGENIC FUSION PROTEIN USING HOMOLOGY MODELING AND SIMULATION BASED APPROACH
Objective: AF9-MLL has been implicated in the pathogenesis of AML, New Therapeutic regimens are prerequisite for this category of hematological malignancy due to the poor prognosis. The experimental 3D structure of AF9-M...