Anti-inflammatory Effects of S. cumini Seed Extract on Gelatinase-B (MMP-9) Regulation against Hyperglycemic Cardiomyocyte Stress

Figure 2

Effect of S. cumini methanol seed extract (MSE) on gelatinolytic activity. (a) Gel-diffusion assay: gelatinolytic activity was measured by digested zones of gelatin around the wells. Standard curve of trypsin was plotted (enzyme units), and enzyme activity for the samples was calculated from the standard graph of trypsin and represented as histogram. The width of the digested zone is proportional to the extent of substrate cleavage and can be used to quantitate protease activity. After giving treatment, we compared our results with trypsin digested zones and found the gelatinase activity in trypsin units. (b) Gelatin zymography: to depict the MSE effect on matrix metalloproteinase (MMP) enzyme activity. Lane 1: control; Lane 2: glucose induced (GI); Lane 3: glucose induced+S. cumini methanol seed extract (GI+MSE) treated; Lane 4: MSE-treated alone (MSE); Lane 5: glucose induced+gallic acid treated (GI+GAL); Lane 6: gallic acid alone (GAL) (). (c) Cell in situ zymography: fluorescence micrographs showing the increase in gelatinolytic activity in HG-induced cells whereas S. cumini methanol seed extract (MSE) and gallic acid (GAL) treatment of HG-glucose stressed cells reverses the activity near the control. Images captured at 40x magnification (). Scale bar 20 μm.

(a)

(b)

(c)