Effects of EGCG in antioxidative enzyme gene expressions and activities in short term treatment.. Young PDL20-30 HDF was treated with 25 and 50μM EGCG for 24 hours and then the antioxida
Trang 1Figure 18 Effects of EGCG in antioxidative enzyme gene expressions and activities in short term treatment
Young (PDL20-30) HDF was treated with 25 and 50μM EGCG for 24 hours and then the antioxidative enzyme gene expressions (A) were measured by quantitative real-time PCR and the enzyme activities (B) were determined by Cayman chemical kits, as mentioned in Section 3.2.7 and 3.2.8, respecctively The data shown are the mean from 3 independent experiments, #p<0.05, comparing untreated HDF-Yong (Y) with untreated HDF-Old (O); *p<0.05, comparing treated HDF-Y with untreated HDF-Y, and +p<0.05 comparing treated HDF-O with untreated HDF-O, respectively; as determined by one-way ANOVA
Trang 2Effects of EGCG in long term treatment
In the long term treatment, the middle-aged HDF (PDL35) was continuously cultured in MEM with 12.5 μM of EGCG until they reach replicative senescence
In the process of HDF reaching the end point, samples were collected, and intracellular ROS accumulation, mitochondrial integrity, antioxidative enzyme activities and gene expressions were examined and compared It was found that middle-aged HDF treated with 12.5 μM EGCG for long term decreased the intracellular ROS level significantly (P<0.05) (Figure 19A) and in the meantime increased the mitochondrial membrane potential by 61.5 % compared to the control (Figure 19B) Moreover, the long term incubation of middle-aged HDF with 12.5 μM of EGCG significantly (P<0.05) increased mitochondrial DNA integrity, which was demonstrated by a much higher mtDNA/nDNA ratio in the treated group for both D310 and D-loop region (Figure 19C) In addition, upon EGCG (12.5 μM) treatment on middle-aged HDF, all of the gene expressions of CAT, SOD1, SOD2 and GPx extensively increased by 58.3, 86.7, 73.8 and 94.0%, respectively (Figure 19D) Besides, the enzyme activities of CAT, SOD1 and SOD2 also enhanced significantly by 11.6%, 58.0 and 47.9%, respectively, whereas GPx activity dropped by about 18.9 % (Figure 19E)
Trang 3Figure 19 Effects of EGCG on the middle-aged HDF in long term treatment
HDF (PDL35) incubated with 12.5 μM EGCG for long term was tested for intracellular ROS (A), mitochondrial potential (B), mitochondrial DNA integrity (C), antioxidative enzyme gene expressions (D) and enzyme activities (E) as mentioned in Materials and methods; *p<0.05, compared to the untreated group as determined by the Student’s t-test; MFI (median fluorescence intensity) was used
as an indicator of the ROS level (C) and (D) show the normalized results where the values of EGCG untreated HDF is set as ‘one’