The effects of chlorogenic acid on serum oxidative stress/antioxidant parameters changed by potassium dichromate
The effects of potassium dichromate, chlorogenic acid, and the combination of these on serum oxidative stress/antioxidant parameters are presented in Table 1. No significant difference was found between the control group (Group I) and all the other groups in respect of serum MDA levels and SOD and GPx activity (p>0.05).
Table 1
The serum oxidant-antioxidant parameters of all the groups.
| Group 1 (n=6) | Group 2 (n=6) | Group 3 (n=6) | Group 4 (n=6) | Group 5 (n=6) | Group 6 (n=6) | P value |
MDA nmol/ml Median(IQR) | 1,175± 0,13 | 1,200±0,12 | 1,060± 0,220 | 1,085± 0,150 | 1,125± 0,16 | 1,130± 0,110 | 0,144 |
GSH mg/L Median(IQR) | 384,830± 141,08bcd | 185,375 ±70,47a | 168,040± 376,300ad | 299,485± 170,700ac | 396,195± 137,85bcd | 458,240± 219,39b | 0,014 |
SOD ng/ml Median(IQR) | 2,075± 0,28 | 2,160 ±0,130 | 2,030± 0,120 | 2,140± 0,170 | 2,065± 0,37 | 1,915± 0,10 | 0,091 |
CAT ng/ml Median(IQR) | 29,005± 6,84b | 34,290 ±2,39bc | 33,690± 5,730bc | 37,825± 2,110ad | 35,265± 7,90bd | 35,580± 7,20acd | 0,023 |
GPX ng/ml Median(IQR) | 17,35± 11,90 | 16,875 ±12,13 | 16,070± 3,330 | 15,405± 6,200 | 13,735± 2,93 | 13,28± 0,53 | 0,067 |
MDA: Malondialdehyde, GSH: glutathione, SOD: superoxide dismutase, CAT: catalase, GPx: glutathione peroxidase, |
In the comparison of the GSH levels in all the groups, with the exception of a significant decrease in Group I compared to the potassium dichromate group (Group II) (p>0.05), no other statistically significant difference was determined (p>0.05). In the comparison of all the treatment groups with the potassium dichromate group, there was a significant increase in GSH levels in the groups given chlorogenic acid at the highest dose alone (Group VI) or before potassium dichromate (Group V), and the GSH levels were observed to have reached those of the control group (p<0.05).
In the comparisons of the control group with all the other groups in respeect of CAT activity, with the exception of the group administered 20mg/kg CGA before potassium dichromate (Group IV) and the group given 40mg/kg CGA alone (Group VI), no other significant difference was observed (p>0.05). When all the treatment groups were compared with the potassium dichromate group, a significant increase was observed in CAT activity only in the group administered 20 mg/kg CGA before potassium dichromate (Group IV) (p<0.05).
Table 1. The serum oxidant-antioxidant parameters of all the groups.
The effects of chlorogenic acid on serum proinflammatory cytokine levels changed by potassium dichromate
The effects of potassium dichromate, chlorogenic acid, and the combination of these on serum proinflammatory cytokine levels are presented in Figure 1. No significant change was determined in serum TNF-α levels when the control group (Group I) was compared with all the other groups (p>0.05) (Figure 1a). No statistically significant difference was determined in the IL-1β levels between Group I and the group given potassium dichromate (Group II) (p>0.05).
When all the treatment groups (Groups III-VI) were compared with Group II, there was observed to be a significant decrease in the IL-1β levels of the group given CGA only at the highest dose (Group VI) and of the group administered 40mg/kg CGA before potassium dichromate (Group V) (p<0.05) (Figure 1b). Compared with the control group (Group I), there was seen to be a significant increase in the IL-6 level of the potassium dichromate group (Group II) (p<0.05). In the comparisons between the potassium dichromate group and all the CGA-administered groups (Groups III-V), no statistically significant difference was determined (p>0.05) (Figure 1c).
Figure 1. The serum proinflammatory cytokine levels of all the groups
The effects of chlorogenic acid on the oxidative stress/antioxidant parameters of kidney tissue changed by potassium dichromate
The effects of potassium dichromate, chlorogenic acid, and the combination of these on kidney tissue oxidative stress/antioxidant parameters are presented in Table 2. No significant difference was found between the control group (Group I) and all the other groups in respect of kidney tissue MDA levels and CAT activity (p>0.05).
Table 2
The kidney oxidant-antioxidant parameters of all the groups
| Group 1 (n=6) | Group 2 (n=6) | Group 3 (n=6) | Group 4 (n=6) | Group 5 (n=6) | Group 6 (n=6) | P value |
MDA nmol/ml Median(IQR) | 1,745± 0,23 | 1,560 ±0,15 | 1,560± 0,14 | 1,730± 0,360 | 1,620± 0,35 | 1,675± 0,34 | 0,254 |
GSH mg/L Median(IQR) | 5,500± 25a | 324,645 ±82,0b | 7,300± 1,8a | 5,00± 0,5a | 5,000± 0,50a | 29,155± 104,7a | 0,002 |
SOD ng/ml Median(IQR) | 3,270± 0,95a | 3,280 ±0,67a | 2,660± 0,210bc | 2,610± 0,140b | 2,875± 0,46ab | 2,905± 0,43ac | 0,002 |
CAT ng/ml Median(IQR) | 45,065± 7,99 | 44,160 ±3,70 | 42,030± 2,13 | 44,140± 6,37 | 40,490± 7,89 | 42,655± 5,56 | 0,495 |
GPX ng/ml Median(IQR) | 27,495± 2,20a | 27,415 ±2,60a | 30,260± 10,30a | 26,815± 7,80a | 23,300± 2,30b | 30,670± 4,90a | 0,025 |
MDA: Malondialdehyde, GSH: glutathione, SOD: superoxide dismutase, CAT: catalase, GPx: glutathione peroxidase, |
In the comparisons between the control group (Group I) and all the other groups in respect of GSH levels, with the exception of a significant increase in the potassium dichromate group (Group II) (p<0.05), no other significant difference was determined (p>0.05).
In the comparisons between the control group (Group I) and all the other groups in respect of SOD activity, a significant difference was determined only in the groups administered 10mg/kg CGA (Group III) and 20mg/kg CGA (Group IV) before potassium dichromate (p<0.05). With the exception of Groups III and IV, the SOD activity of the other treatment groups (Groups V and VI) was seen to be at the same level as that of the control group (Group I) and the potassium dichromate group (Group II) (p>0.05).
In the comparisons between the control group (Group I) and all the other groups in respect of GPx activity, with the exception of a significant decrease in the group administered 40mg/kg CGA before potassium dichromate (Group V) (p<0.05), no other significant difference was determined (p>0.05).
Table 2. The kidney oxidant-antioxidant parameters of all the groups
The effects of chlorogenic acid on the kidney proinflammatory cytokine levels changed by potassium dichromate
The effects of potassium dichromate, chlorogenic acid, and the combination of these on the kidney tissue proinflammatory cytokine levels are presented in Figure 2. No significant difference was determined between the control group (Group 1) and all the other groups in respect of kidney tissue TNF-α, IL-1β, and IL-6 levels (p>0.05) (Figure 2a, b, c).
Figure 2. The kidney proinflammatory cytokine levels of all the groups
The effects of chlorogenic acid on the oxidative stress/antioxidant parameters of liver tissue changed by potassium dichromate
The effects of potassium dichromate, chlorogenic acid, and the combination of these on the liver tissue oxidative stress/antioxidant parameters are presented in Table 3. No significant difference was found between the control group (Group I) and all the other groups in respect of liver tissue MDA-GSH levels and SOD and GPx activity (p>0.05).
Table 3
The liver tissue oxidant-antioxidant parameters of all the groups
| Group 1 (n=6) | Group 2 (n=6) | Group 3 (n=6) | Group 4 (n=6) | Group 5 (n=6) | Group 6 (n=6) | P value |
MDA nmol/ml Median(IQR) | 1,485± 0,26 | 1,32 ±0,43 | 1,060± 0,220 | 1,30± 0,55 | 1,38± 0,24 | 1,39± 0,54 | 0,376 |
GSH mg/L Median(IQR) | 18,70± 64 | 194,50 ±241 | 154,26± 129 | 144,24± 254 | 199,98± 36 | 21,22± 238,0 | 0,060 |
SOD ng/ml Median(IQR) | 2,480± 0,46 | 2,265 ±0,38 | 2,145± 0,23 | 2,28± 0,30 | 2,245± 0,390 | 2,30± 0,32 | 0,237 |
CAT ng/ml Median(IQR) | 44,28± 5,0b | 26,95 ±5a | 27,90± 4,0ac | 30,80± 9,0cd | 29,44± 3,0acd | 34,47± 8,0 bd | 0,001 |
GPX ng/ml Median(IQR) | 23,165± 2,76 | 18,69 ±8,73 | 16,62± 3,70 | 18,60± 6,74 | 20,09± 8,56 | 19,29± 6,76 | 0,153 |
MDA: Malondialdehyde, GSH: glutathione, SOD: superoxide dismutase, CAT: catalase, GPx: glutathione peroxidase, |
In the comparisons related to CAT activity, with the exception of Group VI, a significant decrease was observed in all the other groups compared to the control group (Group I) (p<0.05). When all the treatment groups were compared with the potassium dichromate group, a significant increase was determined in CAT activity only in the group administered 20mg/kg CGA before potassium dichromate (Group IV) (p<0.05).
Table 3. The liver tissue oxidant-antioxidant parameters of all the groups
The effects of chlorogenic acid on liver tissue proinflammatory cytokine levels changed by potassium dichromate
The effects of potassium dichromate, chlorogenic acid, and the combination of these on the liver tissue proinflammatory cytokine levels are presented in Figure 3.
When the groups were examined in respect of TNF-α levels, a statistically significant difference was only determined between the control group (Group I) and the potassium dichromate group (Group II) (p<0.05). No statistically significant difference was determined between Group II and all the CGA-administered groups (Groups III-V) (p>0.05) (Figure 3a).
A significant decrease was seen in the IL-1β levels of all the groups compared to the control group (Group I) (p<0.05).
No statistically significant difference was determined between the potassium dichromate group and all the CGA-administered groups (Groups III-V) (p>0.05) (Figure 3b).
No significant difference was determined was determined between the control group (Group I) and all the other groups in respect of IL-6 levels (p>0.05) (Figure 3c).
Figure 3. The liver tissue proinflammatory cytokine levels of all the groups
When the histopathological semi-quantitative scores of the kidney and liver tissues were evaluated, statistically high levels of damage in the kidney tubulointerstitial and glomerular areas and in the liver tissue were determined only in the potassium dichromate group (Group II) compared to the control group (Group I) (p<0.05) (Table 4). When all the treatment groups were compared with Group II, the damage in these tissues was determined to have been ameliorated at a significant level only in the group administered 10mg/kg CGA before potassium dichromate (Group III) (p<0.05). As the CGA dose increased, the damage in these areas was determined to be at a mild and moderate level compared with the control group (Table 4).(Figure 4)
Table 4
Semi-quantitative histopathological scoring of the kidney and liver tissues
| | Group I (n=6) | Group II (n=6) | Group III (n=6) | Group IV (n=6) | Group V (n=6) | Grup VI (n=6) | P |
Renal tubulointerstitial space | No damage | 5 | 0 | 1 | 0 | 0 | 0 | <0,001 |
Damage is light | 1 | 0 | 4 | 0 | 0 | 4 |
Damage moderate | 0 | 1 | 0 | 2 | 2 | 1 |
Damage is high | 0 | 5 | 1 | 4 | 4 | 1 |
Renal glomerular area | No damage | 6 | 0 | 2 | 0 | 0 | 0 | <0,001 |
Damage is light | 0 | 0 | 3 | 2 | 0 | 4 |
Damage moderate | 0 | 1 | 1 | 3 | 3 | 2 |
Damage is high | 0 | 5 | 0 | 1 | 3 | 0 |
Liver assessment | No damage | 5 | 0 | 0 | 0 | 0 | 0 | <0,001 |
Damage is light | 1 | 0 | 4 | 2 | 1 | 3 |
Damage moderate | 0 | 2 | 2 | 3 | 3 | 3 |
Damage is high | 0 | 4 | 0 | 1 | 2 | 0 |
Table 4. Semi-quantitative histopathological scoring of the kidney and liver tissues
Figure 4: H&E staining of the kidney and liver tissues of the groups.
When the immunohistochemical semi-quantitative scores of the kidney and liver tissues were evaluated, with the exception of eNOS and NFKB activities in the liver tissue (p>0.05), a statistically sginificant difference was determined in all the other parameters between the control group (Group I) and all the other groups (p<0.05) (Tables 5, 6).
Table 5
Semi-quantitative immunohistochemical scoring of the kidney tissues
| | Group I (n=6) | Group II (n=6) | Group III (n=6) | Group IV (n=6) | Group V (n=6) | Group VI (n=6) | P |
Renal iNOS | No damage | 3 | 0 | 4 | 0 | 0 | 2 | <0,007 |
Damage is light | 3 | 3 | 2 | 5 | 6 | 4 |
Damage moderate | 0 | 3 | 0 | 1 | 0 | 0 |
Damage is high | 0 | 0 | 0 | 0 | 0 | 0 |
Renal eNOS | No damage | 3 | 0 | 6 | 0 | 0 | 1 | <0,001 |
Damage is light | 3 | 3 | 0 | 4 | 5 | 5 |
Damage moderate | 0 | 3 | 0 | 2 | 1 | 0 |
Damage is high | 0 | 0 | 0 | 0 | 0 | 0 |
Renal VEGF | No damage | 0 | 0 | 0 | 0 | 0 | 0 | <0,021 |
Damage is light | 5 | 1 | 6 | 4 | 4 | 6 |
Damage moderate | 1 | 5 | 0 | 1 | 1 | 0 |
Damage is high | 0 | 0 | 0 | 1 | 1 | 0 |
Renal NFKB | No damage | 0 | 0 | 0 | 0 | 0 | 0 | <0,002 |
Damage is light | 5 | 0 | 5 | 1 | 0 | 4 |
Damage moderate | 1 | 2 | 1 | 3 | 5 | 2 |
Damage is high | 0 | 4 | 0 | 2 | 1 | 0 |
iNOS: Inducible nitric oxide synthase, eNOS: endothelial nitric oxide synthase, VEGF: Vascular Endothelial Growth Factor, |
NF-κB: Nuclear factor-κB |
Table 6
Semi-quantitative immunohistochemical scoring of the liver tissues
| | Group I (n=6) | Group II (n=6) | Group III (n=6) | Group IV (n=6) | Group V (n=6) | Group VI (n=6) | P |
Liver iNOS | No damage | 3 | 0 | 0 | 0 | 0 | 3 | <0,001 |
Damage is light | 3 | 2 | 6 | 3 | 3 | 3 |
Damage moderate | 0 | 4 | 0 | 3 | 0 | 0 |
Damage is high | 0 | 0 | 0 | 0 | 3 | 0 |
Liver eNOS | No damage | 0 | 0 | 0 | 0 | 0 | 1 | 0,137 |
Damage is light | 5 | 3 | 6 | 2 | 3 | 5 |
Damage moderate | 1 | 3 | 0 | 4 | 2 | 0 |
Damage is high | 0 | 0 | 0 | 0 | 0 | 0 |
Liver VEGF | No damage | 6 | 0 | 0 | 0 | 0 | 0 | <0,001 |
Damage is light | 0 | 6 | 6 | 5 | 0 | 6 |
Damage moderate | 0 | 0 | 0 | 1 | 5 | 0 |
Damage is high | 0 | 0 | 0 | 0 | 1 | 0 |
Liver NFKB | No damage | 1 | 0 | 1 | 0 | 0 | 0 | 0,590 |
Damage is light | 5 | 6 | 5 | 5 | 5 | 5 |
Damage moderate | 0 | 0 | 0 | 1 | 0 | 1 |
Damage is high | 0 | 0 | 0 | 0 | 1 | 0 |
iNOS: Inducible nitric oxide synthase, eNOS: endothelial nitric oxide synthase, VEGF: Vascular Endothelial Growth Factor, |
NF-κB: Nuclear factor-κB |
When compared with the control group (Group I), a statistically significant difference was only determined in the potassium dichromate group (Group II) in the kidney tissue iNOS (NOS2), eNOS (NOS3), VEGF, and NFKB activities and the liver tissue iNOS (NOS2) and VEGF activities (p<0.05). In the comparison of all the treatment groups (Groups III-V) with the potassium dichromate group (Group II), the damage in these tissues was seen to have been significantly ameliorated only in the group administered 10mg/kg CGA before potassium dichromate (p<0.05) (Tables 5, 6). (Figure 5,6).
Table 5. Semi-quantitative immunohistochemical scoring of the kidney tissues
Figure 5: Immunohistochemical staining of the kidney tissues shows the changes
Table 6. Semi-quantitative immunohistochemical scoring of the liver tissues
Figure 6: Immunohistochemical staining of the liver tissues shows the changes