Effect of bone marrow mesenchymal stem cells-conditioned medium on the apoptosis of INS-1 cells
10.11855/j.issn.0577-7402.2015.05.01
- Author:
Kun ZHAO
1
Author Information
1. Department of Endocrinology, General Hospital of PLA
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Inflammation;
INS-1 cells;
Mesenchymal stem cells
- From:
Medical Journal of Chinese People's Liberation Army
2015;40(5):339-343
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect and mechanism of bone marrow mesenchymal stem cell-conditioned medium (BMSC-CM) on pro-inflammatory cytokines-induced apoptosis of insulinoma cells (INS-1). Methods-The conditioned medium containing various cytokines and growth factors was collected and condensed from the supernatants of culture medium of BMSC of rats at passage three. After exposed to the medium with a pro-inflammatory cytokines cocktail containing TNF-α, IL- 1β and IFN-γ for 48 hours, INS-1 cells were cultured with BMSC-CM in different concentrations for another 12 hours. INS-1 cells were divided into the following five groups: control group, pro-inflammatory cytokines-treated group, 1× BMSC-CM-treated group, 2× BMSC-CM-treated group and 4× BMSC-CM-treated group. The survival rate of INS-1 cells was assessed by CCK8 assay. The incidence of cell apoptosis was measured using Annexin V/PI staining. Intracellular reactive oxygen species (ROS) production was determined with dichlorofluorescein diacetate (DCFH-DA). Results-Compared with the control group, prolonged exposure to pro-inflammatory cytokines induced significant decrease in the viability of INS-1 cells (75.84%±1.53%) and a great increase in the incidence of cell apoptosis (17.23%±1.77%), and also in intracellular ROS level (34.3%±0.80%) with statistically significant difference (P<0.01). However, BMSC-CM treatment protected INS-1 cells against pro-inflammatory cytokines-induced injury in a dose-dependent manner. Among the BMSC-CM of three different concentrations, 4× BMSC-CM exhibited best protective effects on pro-inflammatory cytokines-treated INS-1 cells, as reflected by increased cell viability (87.7%±2.08%) and significant reduction in the incidence of cell apoptosis (8.67%±1.59%), and also in intracellular ROS production (17.1%±2.14%) with statistically significant difference (P<0.05). Conclusion-BMSC-CM protected INS-1 cells against pro-inflammatory cytokines-induced apoptosis, the possible mechanism underlying this effect might be associated with the reduction of intracellular ROS level.
