# The Effects of _Anthyllis vulneraria_ Hydroalcoholic Leaf Extract as an Adjuvant in Wound Healing
## 3 Results
### Experimental Animal Study
#### 3.2.2 Oxidative Stress Markers Evaluation  
The Shapiro-Wilk test indicated that the data were normally distributed amongst the groups (\(p>0.05\)). The analysis of oxidative stress markers in the tissue samples revealed significant differences between the experimental groups, while the MDA levels in blood showed no significant variation (\(p=0.966\)). Detailed levels of the investigated markers are illustrated in Figure 6.  
Notably, MDA levels in the tissue samples were significantly lower in the EG2 group compared to the NC (\(p<0.001\)). Additionally, a trend toward significance was observed between EG1 and the NC (\(p=0.066\)). GSH levels were significantly elevated in both experimental groups compared to the NC (both \(p<0.001\)), with EG2 displaying the highest values among all groups. Similarly, SSSG levels were significantly reduced in both EG1 and EG2 relative to the NC (\(p\leq 0.001\)), with EG2 also showing a significant decrease compared to the PC (\(p=0.035\)). Moreover, SSSG levels differed significantly between EG1 and EG2 (\(p=0.017\)). The GSH/GSSG ratio followed a consistent pattern, with both experimental groups differing significantly from the NC (both \(p<0.001\)). Furthermore, EG2 exhibited a significantly higher ratio compared to both the positive control and EG1 (both \(p<0.001\)). The highest ratio was recorded in EG2, followed by EG1, the PC, and the NC. Among the oxidative stress markers analyzed, CAT activity remained unchanged across all groups except for the PC, which exhibited an increase compared to the others. SOD activity was significantly elevated in both experimental groups compared to the NC (both \(p\leq 0.001\)), with the highest levels observed in EG2, followed closely by EG1 and then by PC. A similar trend was noted for GPX activity, which reached its peak in the experimental groups, particularly EG2, with significantly higher values than the NC (\(p=0.011\) for EG1 and \(p=0.006\) for EG2). IL-8 values were significantly different between the groups, presenting smaller levels in the experimental groups. There were differences between both experimental groups compared to the NC (\(p=0.002\) and \(0.017\), respectively) and PC (\(p=0.004\) and \(0.35\), respectively). The lowest value was in EG1 followed by EG2. These findings suggest that the experimental treatments effectively modulated oxidative stress markers and IL-8 secretion, particularly in the EG2 group, which consistently displayed the most pronounced differences across multiple parameters. In all the parameters tested, EG2 showed either the highest or the lowest value, except for CAT, whose value in the EG2 group was not statistically different from the controls, and for IL-8, where EG1 had the lowest value. The highest values of EG2 were determined for GSH, GSH/GSSG, SOD, and GPX, while the lowest values of EG2 were for MDA and  
Figure 6: Oxidative stress markers of _A. vulneraria_ hydroalcoholic leaf extract cream in the skin samples of the four groups: NC (Negative Control, sample size = 8), PC (Positive Control, sample size = 8), EG1 (Experimental Group 1, sample size = 8), and EG2 (Experimental Group 2, sample size = 8): (a) malondialdehydeehyde (MDA) levels (nmol/mg protein); (b) reduced glutathione (GSH) levels (nmol/mg protein); (c) oxidized glutathione (GSSG) levels (nmol/mg protein); (d) GSH/GSSG levels; (e) catalase (CAT) levels (U/g protein); (f) superoxide dismutase (SOD) levels (U/g protein); (g) glutathione peroxidase (GPX) levels (U/g protein); and (h) interleukin 8 (IL-8) levels (pg/mg protein). * \(p<0.05\) and **\(p<0.001\)**, all groups vs. NC; # \(p<0.05\) and **\(p<0.001\)** vs. PC. GSSG. EG1 followed the same pattern as EG2, but with slightly lower or higher values, except for IL-8, where the lowest levels were found in EG1. EG2 exhibited an ability to increase the antioxidant capacity in wound tissue, which was demonstrated by low values of MDA and GSSG and increased levels for endogenous antioxidants--both non enzymatic and enzymatic--such as GSH, GSH/GSSG, SOD, and GPX.