Antioxidant and Anti-inflammatory Effect of Black Garlic Aqueous Extract on LPS induced Vero cell

Amna Mohamed Abdulhafith Marie, Dr.biol.hom. Nastiti Wijayanti, M.Si

2020 | Tesis | MAGISTER BIOLOGI

Inflammation is the process that occurs in the body as an immune response due to risk stimulate. Serious damage can occur if the inflammatory response exceeds the protective anti-inflammatory effects. Black garlic (BG) is one of the natural resources that has been reported can be used as an anti-inflammatory agent. This research aimed to study the effect of aqueous extract of Black garlic as an anti-inflammatory. The methods of this research were used the extraction of BG by maceration method using aqueous as a solvent, S-allylcysteine as a potential compound was tested using HPLC technique. Antioxidant properties of BG was tested by using DPPH and H2O2 scavenging assays. Cytotoxicity test was done with WST-1 assay in 96-well plate. The anti-inflammatory effects of BG were evaluated in the lipopolysaccharide (LPS)- induced Vero cell. Nitric Oxide (NO) production was determined by the Griess assay, while expression of TNF-alpha, iNOS and COX-2 in mRNA and protein levels were measured using RT-PCR and Western blot analysis. The result showed that BG extract contains S-allylcysteine and has an ability to scavenging free radical activities. The BG aqueous extract did not show cytotoxicity on Vero cells up to 750µg/ml. LPS-induced nitric oxide production in Vero cell was reduced after BG treatment, as well as the results of RT-PCR showed that BG markedly inhibited LPS-stimulated iNOS and COX-2 gene and protein expression, as well as TNF-alpha. This study concluded that the administration of aqueous extract of Black Garlic has the potential as an antioxidant and anti-inflammatory by inhibiting the expression of NO, iNOS and COX-2 mRNAs.

Inflammation is the process that occurs in the body as an immune response due to risk stimulate. Serious damage can occur if the inflammatory response exceeds the protective anti-inflammatory effects. Black garlic (BG) is one of the natural resources that has been reported can be used as an anti-inflammatory agent. This research aimed to study the effect of aqueous extract of Black garlic as an anti-inflammatory. The methods of this research were used the extraction of BG by maceration method using aqueous as a solvent, S-allylcysteine as a potential compound was tested using HPLC technique. Antioxidant properties of BG was tested by using DPPH and H2O2 scavenging assays. Cytotoxicity test was done with WST-1 assay in 96-well plate. The anti-inflammatory effects of BG were evaluated in the lipopolysaccharide (LPS)- induced Vero cell. Nitric Oxide (NO) production was determined by the Griess assay, while expression of TNF-alpha, iNOS and COX-2 in mRNA and protein levels were measured using RT-PCR and Western blot analysis. The result showed that BG extract contains S-allylcysteine and has an ability to scavenging free radical activities. The BG aqueous extract did not show cytotoxicity on Vero cells up to 750µg/ml. LPS-induced nitric oxide production in Vero cell was reduced after BG treatment, as well as the results of RT-PCR showed that BG markedly inhibited LPS-stimulated iNOS and COX-2 gene and protein expression, as well as TNF-alpha. This study concluded that the administration of aqueous extract of Black Garlic has the potential as an antioxidant and anti-inflammatory by inhibiting the expression of NO, iNOS and COX-2 mRNAs.

Kata Kunci : Black garlic, Vero cell, anti-inflammatory, TNF alpha, iNOS, COX-2.