To quantify the rate of superoxide anion production over time in response to the different treatment conditions, sigmoidal and linear equations were fitted to the data (See Materials & Methods). Figure 3b consists of the data fitted to the sigmoidal equation for AA and combination treatment, and linear equation for control, metformin, and sulindac treatments. The nonlinear fitting used for AA and the combination treatment demonstrates a greater increase in the O2•− production rate in the aforementioned groups in comparison to the remaining groups. This can be further justified by Table 1, where slopes of AA and the combination of metformin and sulindac are notably greater than the linear slopes. Table 2 depicts the percent change between the slopes of the control group and the treatment groups, including AA. The percent change between the control and positive control group is the greatest (75.93%), followed by the difference between the control and combination of metformin and sulindac groups (33.22%), control group and sulindac (7.88%), and lastly control group and metformin (5.48%). The highest difference was expected, as AA is the positive control, though the greater difference in percent change between control and combination group, compared to control and the treatments alone indicates the role of the drug combination in superoxide production increases over time.
The slopes of red channel fluorescence intensity for each group are compared to one another statistically in Figure 4 and showed significant changes in the positive control and combined treatment compared to the control group. In these treatment cases, there was a large difference in the rate of superoxide production over time with respect to the control group, reflecting the effects of the drugs on the mitochondria and its processes.