Cucumis sativus L. Seeds Ameliorate Muscular Spasm-Induced Gastrointestinal and Respiratory Disorders by Simultaneously Inhibiting Calcium Mediated Signaling Pathway

Abstract

Cucumis sativus L. is globally cultivated as an edible vegetable. Besides its nutritional benefits, it is used in traditional medicines against various ailments. The current study was designed to elucidate the multi-target mechanisms of a C. sativus seeds extract against asthma and diarrhea using network pharmacology along with a molecular docking approach. Furthermore, in-vitro and in-vivo experiments were conducted to verify the mechanistic insight of in silico studies. LC-ESI-MS/MS was performed to identify the bioactive compounds in the extract; later, some compounds were quantified by HPLC. C. sativus seed. EtOH has kaempferol in higher concentration 783.02 µg/g, followed by quercetin (693.83 µg/g) and luteolin (617.17 µg/g). In silico studies showed that bioactive compounds interfered with asthma and diarrhea-associated target genes, which are members of calcium-mediated signaling to exert a calcium channel blocker activity. The seeds extract exerted a concentration-dependent spasmolytic response on isolated jejunum, trachea, and urinary bladder preparations and caused relaxation of spastic contraction of K+ (80 mM) with suppressed calcium concentration-response curves at dose 0.3 and 1 mg/mL. It also showed antiperistalsis, antidiarrheal and antisecretory activity in animal models. Thus, C. sativus seeds have therapeutic effects by regulating the contractile response through a calcium-mediated signaling pathway.

Antidiarrheal and Cardio-Depressant Effects of Himalaiella heteromalla (D.Don) Raab-Straube: In Vitro, In Vivo, and In Silico Studies

“…The separation factor and resolution of all separated compounds were >1.0 and >1.5, respectively. The reproducibility of separate components was also good with RSD < 2% (run to run) and 2.7% (day to day) [ 12 ]. The HPLC chromatograms for the identified phenolic and flavonoid compounds are shown in Figure 1 , and Table 1 presents the phenolic and flavonoid compounds identified in Hh.Cr.…”

“…The crude extract (Hh.Cr) and its ethyl acetate (Hh.Ea) fraction caused the negative chronotropic effect (i.e., decrease in heart rate) and negative inotropic effect (i.e., force of contraction) on atrium preparation [ 12 ]. Hh.Cr and its Hh.Ea fraction showed negative inotropic effect within concentration range 0.01–5.0 mg/mL with EC 50 0.…”

“…The docking calculations are beneficial to predict ligand pose within the binding site of the target protein. The involvement of physical energies terms (i.e., solvation energy) with suitable force field make docking calculation of compounds more acceptable with accuracy ( Table 2 ) [ 12 , 13 , 14 ].…”

“…The mobile phase consisted of solvent A (methanol (30): acetonitrile (70)) and solvent B (0.5% glacial acetic acid in double-distilled water), mobile phase run at flow rate 0.08 mL/min, and data was recorded at 275 nm of UV spectra. The peaks and retention times of phenolic acids and flavonoids of H. heteromalla were matched with external standards to quantify the components [ 12 ]. The resolution and separation factor was used to determine HPLC separation efficiency.…”

“…The antidiarrheal activity was performed according to the method prescribed by Wahid et al [ 12 ] with modifications. Mice (20) of either sex were divided into five groups, i.e., negative control (0.9% normal saline), standard drug (loperamide 10 mg/kg), and H. heteromalla doses (100, 200, and 400 mg/kg).…”