UPLC-qTOF-MS Phytochemical Profile and Antiulcer Potential of Cyperus conglomeratus Rottb. Alcoholic Extract

Abstract

Cyperus has been commonly used as a multi-use medicinal plant in folk medicine worldwide. The objectives of our study were to determine the different metabolites in the Cyperus conglomeratus Rottb. methanol extract, and to assess its in vivo gastroprotective effect in ethanol-induced gastric ulcer model in rats. Serum levels of galactin-3 and TNF-α were employed as biochemical markers. To pinpoint for active agents, comprehensive metabolites profiling of extract via UPLC-qTOF-MS/MS was employed. A total of 77 chromatographic peaks were detected, of which 70 were annotated. The detected metabolites were categorized into phenolic acids and their derivatives, flavonoids, stilbenes, aurones, quinones, terpenes, and steroids. Rats were divided into six groups; healthy control, ulcer control, standard drug group, and 25, 50, 100 mg/kg of C. conglomeratus treated rats. Pre-treatment with C. conglomeratus alcohol extract significantly reduced galactin-3, and TNF-α in ethanol-induced ulcer model at 25, 50, and 100 mg/kg. Further histopathological and histochemical studies revealed moderate erosion of superficial epithelium, few infiltrated inflammatory cells, and depletion of gastric tissue glycoprotein in the ulcer group. Treatment with the extract protected the gastric epithelial cells in a dose-dependent manner. It could be concluded that C. conglomeratus extract provides significant gastroprotective activity in ethanol-induced gastric ulcer and ought to be included in nutraceuticals in the future for ulcer treatment.

Keywords: Cyperus conglomeratus; UPLC-qTOF-MS; biochemical and histochemical characteristics; gastroprotective activity; metabolite profiling.

Figure 1

UPLC-qTOF-MS base peak chromatogram of C. conglomeratus alcoholic extract in negative ionization mode. Peak numbering follows that stated in Table 1.

Figure 2

Representative structures of major metabolites identified in C. conglomeratus extract.

Figure 3

Anti-ulcer activity of 70% alcoholic extract of above ground of C. conglomerates; (A) normal control showing stomach mucosa with intact surface mucosal epithelium and no lesion appeared; (B) stomach mucosa of an ulcer group showing severe disruption of the surface epithelium and necrotic lesions mucosa with inflammatory cells infiltration and hemorrhage as well; (C) stomach mucosa of rat treated with 25 mg/kg. b.w of 70% alcoholic C. conglomeratus extract showing mild mucosal surface erosion mild disruption to the surface epithelium mucosa with no edema and no leucocytes infiltration of the submucosal layer; (D) stomach mucosa of treated with 50 mg/kg. b.w of 70% alcoholic C. conglomeratus extract showing mild erosion; (E) mucosa of group treated with 100 mg/kg. b.w of 70% alcoholic C. conglomeratus extract shows intact surface mucosal epithelium and no lesion appeared; (F) mucosa of group treated with Ranitidine showing mild erosion of mucosa. The scale bar length is 200 µm.

Figure 4

The effects of 70% alcoholic extract of above ground of C. conglomeratus on gastric tissue glycoprotein in ethanol-induced gastric ulcers in rats. (A) Normal control showing no accumulation of the magenta color in the mucosal cell layer; (B) stomach mucosa of ulcer group showing no accumulation of the magenta color in the mucosal cell layer; (C, D, and E, respectively): 25, 50, and 100 mg/kg. b.w of 70% alcoholic C. conglomeratus extract showing an increase in magenta color in the mucosal cell layer compared to the ulcerated group in a dose-dependent manner; (F) rat treated with Ranitidine showing an increase in magenta color in the mucosal cell layer compared to the ulcerated group (PAS stain, scale bar; 200 µm).