NEPHROPROTECTIVE EFFECT AND MECHANISM OF ACTION OF ETHANOLIC EXTRACTS OF MADEIRA VINE (ANREDERA CORDIFOLIA (TEN.) STEENIS LEAVES, ETHANOLIC EXTRACTS OF SOWTHISTLE (SONCHUS ARVENSIS L.) LEAVES AND ITS COMBINATIONS IN ANIMAL KIDNEY FAILURE MODELS

• Main Author: Suliska, Nova
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/66214
• Institution: Institut Teknologi Bandung

Renal failure is characterized by decreased glomerular filtration rate (less than 60 mL/min per 1.73 m2), albuminuria, increased serum creatinine levels, serum urea nitrogen levels, abnormalities in renal histology, electrolytes, or other abnormalities due to tubular damage. Renal failure is caused by glomerulonephritis and nephrotoxic drugs such as aminoglycosides and nonsteroidal anti-inflammatory drugs. Until now, no medication works specifically to treat kidney failure. Binahong showed the effect of improving kidney function by lowering serum creatinine levels and serum urea levels, while tempuyung was shown to have a diuresis and vasodilatation effect in rats. This study aimed to examine the nephroprotective effect and mechanism of the ethanolic extract of binahong leaves, tempuyung leaves and the best combination dose of ethanolic extracts of binahong leaves and tempuyung leaves in providing a nephroprotective effect. The ethanol extract of binahong leaves and tempuyung leaves was extracted by reflux using 96% ethanol as solvent. The yield of ethanol extract of binahong leaves was 9.76%, while that of tempuyung leaves was 9.69%. The results of TLC and HPLC analysis showed that the flavonoid content of vitexin was 12.06 ± 0.13 g/mg in the ethanol extract of binahong leaves, luteolin was 3.12 ± 0.04 g/mg, and luteolin-7-O-glucoside was 3,02 ± 0.08 µg/mg on an ethanol extract of tempuyung leaves. Nephroprotective test results in animal kidney failure by gentamicin and piroxicam show doses of 100 mg/kg b.w can reduce serum urea levels by 25-30%, serum creatinine levels by 16-23%, decrease urine volume, kidney index, TBARS levels, and significantly increased catalase activity against the positive control group (p<0.05). The combination of ethanol extract of binahong leaves and tempuyung leaves consists of several combination doses to see the effect of the extract which is more dominant, namely binahong 25 mg/kg b.w + tempuyung 75 mg/kg b.w (Bin 25 + Temp 75), binahong 50 mg/kg b.w + tempuyung 50 mg/kg b.w (Bin 50 + Temp 50), v dan binahong 75 mg/kg b.w + tempuyung 25 mg/kg b.w (Bin 75 + Temp 25) and compared to the single extracts, namely binahong 100 mg/kg b.w (Bin 100) and tempuyung 100 mg/kg b.w (Temp 100). The combination groups did not show a significant difference in the reduction of serum urea and creatinine levels compared to the single extract group. The effect of the combination group was comparable to the effect of a single extract. The Bin 75 + Temp 25 group showed the best decrease in serum urea and creatinine levels. The kidney of induced test animals was pale in colour and larger in size than normal kidneys. In the Bin 75 + Temp 25 group, the colour of the kidneys started to turn red, similar to that of the normal group. The decrease in kidney index occurred by 0.7-18% in the group given the extract either alone or in combination. Binahong, tempuyung extract, and their combination can reduce Kidney injury molecule 1 (KIM-1) and N-acetyl-?-D-glucosaminidase (NAG) levels significantly compared to positive controls as specific markers of kidney tubular damage. The largest decrease in NAG occurred in the Bin 75 + Temp 25 group by 95.6%, and the levels were the same as normal. The antioxidant effect of the ethanol extract of binahong leaves, tempuyung leaves, and their combination reduced nitric oxide (NO) levels by 32-58%. The Bin 75 + Temp 25 group showed the greatest decrease in NO levels. The increase in SOD activity shown by the combination was 33-57%, significantly different from the positive control group. The Bin 75 + Temp 25 group showed the lowest levels of TNF- and did not differ significantly from the normal group (P> 0.05). The Bin 75 + Temp 25 group showed activation of ERK 1/2 and p38 which play a role in the growth and proliferation of tubular cells and will trigger the differentiation of CD 24 progenitor cells as seen from the increase in CD 24 expression, which was significantly different compared to the positive control group. The ethanol extract of binahong and leaves, tempuyung leaves, and their combination can also inhibit cell apoptosis mediated by caspase-3. The qPCR results showed a decrease in caspase-3 expression in the Bin 75 + Temp 25 group by 0.33±0.14 (relative to GAPDH expression). The results of the histopathological analysis with Periodic Acid Schiff staining showed that the administration of binahong extract, tempuyung, and their combination could inhibit the damage that occurred in the kidney by reducing the percentage of histopathological kidney damage. The combination of binahong at a dose of 75 mg/kg b.w + tempuyung at 25 mg/kg b.w gave the percentage of kidney damage with the lowest EGTI score. The groups of Bin 100, Bin 75 + Temp 25, Bin 50 + Temp 50, Bin 25 + Temp 75 indicated that brush-border membrane regeneration occurred. The combination group Bin 75 + Temp 25 showed a similar area of BBM to the normal group. Giving ethanol extract of binahong leaves, tempuyung leaves, and their combination can prevent collagen tissue formation. The Bin 75 + Temp 25 group showed a significant decrease in collagen area between Temp 100, Bin 50 + Temp 50, and Bin 25 + Temp 75, but not significantly different against Bin 100. The combination group Bin 75 + Temp 25 showed the best nephroprotective effect in inhibiting collagen tissue formation in animals with kidney failure. vi The combination of ethanol extract of binahong leaf at a dose of 75 mg/kg b.w and tempuyung leaf at a dose of 25 mg/kg b.w showed the best nephroprotective effect compared to the other combination groups. The nephroprotective mechanism of the combination of ethanol extract of binahong leaves and tempuyung leaves is to reduce serum creatinine, urea levels, kidney index, and TNF-?levels, with the mechanism, inhibiting the release of KIM-1, NAG, free radicals and increase SOD activity, increasing cell proliferation by activating the MAPK ERK 1/2 and p38 signalling pathways, increasing CD24 expression for tubular cell differentiation, inhibiting cell apoptosis by decreasing caspase-3 expression, helping tubular cell brush border membrane regeneration, and inhibiting the formation of fibrosis in the kidney.