A comparative study of anti-inflammatory properties and activities of green and red Christia vespertilionis leaves

Christia vespertilionis is a plant traditionally used to treat various diseases, including inflammation. This study aims to compare and analyze the anti-inflammatory properties and activities of green and red C. vespertilionis leaves and to determine the correlation between the protease inhibition assay and phenolic content. Preliminary phytochemical analysis showed the presence of alkaloids, tannins, saponins, cardenolides, phenolics, cardiac glycosides, flavonoids, and steroids in green and red leaves. Green leaf had the highest anti-inflammation potential, with a high extraction yield (6.39 ± 0.01%), phenolic content (29.25 ± 0.50 mg) gallic acid equivalent [GAE]/mL), flavonoid content (1.57 ± 0.03 mg quercetin acid equivalent [QE]/mL) and tannin content (22.70 ± 3.15 mg tannic acid equivalent [TAE]/mL). Green leaf contained more anti-inflammatory compounds, including n-hexadecanoic acid, phytol, 9, 12, 15-octadecatrienoic acid (Z, Z, Z)- and squalene. The protease inhibition assay showed that green leaf had anti-inflammatory activity at 66.55 ± 2.59% with the half-maximal inhibitory concentration (IC50) of 284.59 ± 10.63 μg/mL. The green leaf showed a significant positive correlation between the protease inhibition assay and phenolic content, with R2 = 0.6821. We conclude that both green and red C. vespertilionis types have the potential to act as anti-inflammatory agents, but green leaves are preferable

Optimisation of protease purification from Christia vespertilionis leaves and its anti-inflammatory activity

Christia vespertilionis (L. f.) Bakh. f. is widely known for its anti-inflammatory and anti-cancer properties. However, there are no previous studies about extracting and purifying protease enzymes from C. vespertilionis leaves. Therefore, this study was conducted to extract and optimise the purification of protease from C. vespertilionis leaves and characterise its anti-inflammatory properties. The optimisation was performed using different levels of ammonium sulphate saturation (20, 40, 60, 80 and 100%). Next, dialysis was carried out for the sample with the highest specific activity (16.88 U/mg), achieved with 100% ammonium sulphate saturation. At 100% saturation, C. vespertilionis leaves showed an increase in the specific activity to 20.06 U/mg after dialysis. The findings demonstrate the successful extraction and purification of C. vespertilionis protease (CVP) with a molecular weight of 48 kDa, as proven by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. CVP also exhibited anti-inflammatory activity, with an inhibition of 45.6% in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and IC50 of 19.24 μg/mL. The HPLC test further confirmed the presence of gallic acid and quercetin compounds in C. vespertilionis, which cure inflammation. The results indicate that optimised CVP purification was achieved and its anti-inflammatory ability was proven