Cellular stability was assessed by MTT assay equally as described previously with some modifications. In temporary, after exposing to different concentrations of homocysteine for 24 h, ATP-competitive HCV protease inhibitor the cells were more incubated with the MTT reagent for 4 h at 37uC with five full minutes CO2. Then, DMSO 1 ml was put into reduce farmazan crystals and the OD values were taken at 490 nm by utilizing an Elisa plate reader. Acridine orange/ethidium bromide double staining was used to detect the apoptosis of BMSCs as described previously. BMSCs were fixed with 4% paraformaldehyde for 30 min at room temperature. Then, the cells were stained with Hoechst 333342 for 20 min. After washing twice with serum free DMEM, the cells were resuspended in serum free DMEM for morphological observation using the fluorescence microscope. Viability/Cytotoxicity Assay Kit was used to see live and dead cells. In brief, BMSCs were plated on coverslips and then were handled with different concentrations of homocysteine. The cells were then washed with PBS and stained according to Endosymbiotic theory manufacturers instructions. BMSCs were captured under a fluorescence microscope. The stained live cells display green fluorescence and stained dead cells display red fluorescence. Terminal deoxynucleotidyl transferase dUTP nick finish labeling assay was used to find the effects of homocysteine on BMSCs. The strategy to perform TUNEL assay is just was described previously. BMSCs were fixed with 401(k) paraformaldehyde solution for 1 h at room temperature, and then permeabilized in 0. 1%Triton X 100, followed by freshly prepared TUNEL reaction mixture for 1 h in a dark place. The coverslips were then washed with PBS and observed under a fluorescence microscope. Intracellular ROS degree of BMSCs was quantified by ROS Detection Canagliflozin cell in vivo in vitro Assay Kit. BMSCs were collected and exposed to 10 mM DCFH DA for 20 min at 37uC in a room. After that, BMSCs were washed twice and were then photographed under a fluorescence microscope. Mitochondrial membrane potential was determined using JC 1 probe. Shortly, after treatment with homocysteine for 24 h, BMSCs were stained with 10 mM of JC 1 for 20 min at 37uC. After washing twice with buffer option, BMSCs were examined by using a fluorescence microscope. The procedure to measure VEGF and IGF 1 focus in the culture medium of BMSCs was just as described below. In brief, after BMSCs were treated by homocysteine 30, 100, 300 and 1,000 mM for 72 h, the cultured medium was collected and then centrifuged at 3000 g for 10 minutes. The VEGF and IGF 1 concentration in the supernatants was assayed using VEGF and IGF 1 ELISA systems according to the manufacturers guidelines. The test was performed 3 times. Protein samples were extracted from cultured BMSCs after-treatment with homocysteine. Protein concentration was determined using the BCA method as suggested by the manufacturer. After boiled for 5 min, the protein samples were fractionated by SDS PAGE and transferred to PVDF membrane.