Figure 1.
Effects of different concentrations of T-2 toxin on the cellular viability of chondrocytes were estimated by MTT reduction.
Cells were incubated in absence or presence of several T-2 toxin concentrations for different time periods (1–5 d). *P<0.05.
Table 1.
Values of mitochondrial respiratory chain complexes in cultures of normal chondrocytes treated with 20 ng/ml T-2 toxin for 5 days.
Figure 2.
Effects of T-2 toxin on the mitochondrial membrane potential and ATP.
A. Normal human chondrocytes were exposed to 1 10, 20 ng/ml T-2 toxin and 1 uM Na2SeO3+20 ng/ml T-2 toxin for 5 d. The mitochondrial membrane potential of chondrocytes was analyzed using the fluorescent dye JC-1, and quantified by a flow cytometer. B. Effect of different concentrations of T-2 toxin on the cellular ATP. *P<0.01 vs. control; #P<0.01 vs. 20 ng/ml T-2 toxin group.
Figure 3.
Effects of T-2 toxin on ROS and antioxidants generation.
The production of ROS at mitochondrial (A) and cytoplasmic (B) level was assessed by MitoSOX red mitochondrial superoxide indicator and Carboxy-H2DCFDA probe (both from Invitrogen), respectively. GSH concentration (C) and GPx activity (D) were measured by coupled enzyme assay as described under “Materials and methods”. Chondrocytes were incubated with1, 10, 20 ng/ml T-2 toxin and 1 uM Na2SeO3+20 ng/ml T-2 toxin for 5 d. *P<0.01 vs. control; #P<0.01 vs. 20 ng/ml T-2 toxin group.
Figure 4.
Western blot analysis for cytochrome c protein in normal human chondrocytes after treatment with 1, 10, 20 ng/ml T-2 toxin and 1 uM Na2SeO3+20 ng/ml T-2 toxin for 5 d.
A. Western blot of cytochrome c. B. Increase of cytochrome c relative to control. Data are shown as the mean ± S.D. of at least three separate experiments. *P<0.01 vs. control; #P<0.01 vs. 20 ng/ml T-2 toxin group.
Figure 5.
The effects of T-2 toxin on caspase-9 and caspase-3 activities in human chondrocytes.
Cells were treated with 1, 10, 20 ng/ml T-2 toxin and 1 uM Na2SeO3+20 ng/ml T-2 toxin for 5 d. The activities of caspase-9 (A) and caspase-3 (B) were measured using the substrate Ac-LEHD-pNA and Ac-DEVD-pNA, separately. *P<0.05 vs. control; #P<0.01 vs. 20 ng/ml T-2 toxin group.
Figure 6.
Effect of T-2 toxin on apoptosis in human chondrocytes cultured for 5 days.
The apoptosis rate was determined by FCM assay. *P<0.05 vs. control; #P<0.01 vs. 20 ng/ml T-2 toxin group.