In Vitro and In Silico Evaluation of Betulin on Calcium Oxalate Crystal Formation

Objective: The medicinal plant Betula alba has been used for prevention and treatment of kidney stones. Betulin is one of the main phytochemicals of Betula alba. The aim of this study is to investigate the antioxidant and antiurolithiatic activity of betulin in vitro and in silico. For antioxidant activity, 2, 2-diphenyl-1-picrylhydrazyl (DPPH), total reducing capacity, nitric oxide (NO) radical scavenging assay, and superoxide radical scavenging assay were studied. Method: In order to study antiurolithiatic activity, three assays such as crystallization, nucleation, and aggregation of oxalate crystal in urine were performed. In silico experiments were performed by using AutoDock 4.2 tools in order to establish affinity of phytochemicals toward antioxidant enzyme and matrix metalloproteinase (MMP-2 and 9). Results: The results obtained clearly demonstrate the significant scavenging activity of betulin and cystone against DPPH, NO, and superoxide radicals in comparison to standard antioxidant L-ascorbate (L-AA). It has also been observed that betulin has the capacity to inhibit the crystallization, nucleation, and aggregation in comparison to cystone. On the other hand, betulin and L-AA showed strong affinity toward antioxidant enzymes and matrix metalloproteinase as determined by in silico experiments. Conclusions: From this, it may be concluded that the antiurolithiatic activity of betulin is, at least in part, mediated by its antioxidant property.