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Validation of an Ethnobotanical Process Through In vitro, In vivo and In silico Methods Reveals that Grilling Enhances the Antidiarrheal Activity of Terminalia bellerica Fruits
Submission date: 2017-09-20
So far the ethnobotanical studies have been limited either to survey and documentation or the study of phytocompounds present in medicinal plants. We have validated an “ethnobotanical process”, which can be useful in driving attention of researchers at the process adopted by ethnic tribes for drug preparation. The study can open a new arena in the field of ethnobotany. Tharu and Buksa tribes of Udham Singh Nagar, Uttarakhand, India use grilled fruits of Terminalia bellerica (Gaertn.) Roxb. as an effective cure for diarrhea. We validated the ethnobotanical claim by comparing the antidiarrheal effect of grilled fruits (GF) with dried fruits (DF). The 50% ethanolic extracts of GF and DF were successively fractionated; the antioxidant and bacterial inhibition activity were studied using DPPH free radical scavenging, anti-lipid peroxidation and broth dilution method respectively. Difference in metabolites of ethyl acetate fractions of GF and DF was analyzed using GC-MS, while gallic and ellagic acid content was estimated using HPTLC. Further the in-vivo antidiarrheal effect of ethyl acetate fractions of DF and GF was studied on castor oil induced diarrhea model.The ethyl acetate fractions showed potential DPPH free radical scavenging (IC50 11.13 µg/ml in DF and 8.56 µg/ml in GF), anti-lipid peroxidation and antibacterial activity. The non-targeted metabolic profiling showed higher content of tartaric acid, valeric acid, gallic acid, succinic acid, oxalic acid, malonic acid, malic acid, 1,2,3 trisbenzene, uridine and 11-eicosenoic acid in GF. The HPTLC results indicated that gallic acid content was 2.8 (± 0.14) and 4.92 (±0.28) mg/g while ellagic acid content was 4.7 (±0.32) and 4.45 (±0.45) mg/g dry powder in DF and GF respectively. According to in vivo antidiarrheal activity DF and GF (100 mg/kg oral) inhibited diarrhea by 41.87% and 71.72% respectively. Molecular docking studies using Discovery Studio 4.0 showed that gallic acid could bind to Trp 34 of E chain of the B-oligomer of verotoxin-1 from E.coli. The protein belongs to Shiga toxin family and is associated with diarrheal diseases and uraemic syndrome. Thus our study validated that grilling significantly altered the levels of metabolites in T. bellerica fruits which could be responsible for its increased therapeutic potential. Keywords: Terminalia bellerica, ethnobotanical process, antidiarrheal, gallic acid.