Chemical profiles, pharmacological properties, and in silico studies provide new insights on Cycas pectinata

Abstract

The current study aimed to qualitatively and quantitatively determine the phytochemical components of Cycas pectinata methanol extract (MECP), along with its antioxidant, anti-inflammatory, thrombolytic, locomotor, anxiolytic, analgesic, and antidiarrheal activities. The in vitro antioxidant activity was evaluated by DPPH scavenging assay and the total phenol and total flavonoid contents, while the anti-inflammatory activity was evaluated by a protein denaturation assay. The in vivo locomotor effects were examined using the open field test and hole-cross test. The anxiolytic effect was examined using the elevated plus maze (EPM) test, hole-board test (HBT), and light-dark test (LDT), while the analgesic activity was investigated using the acetic acid-induced writhing test. The antidiarrheal effect was evaluated by castor oil-induced diarrhea and gastrointestinal motility. Ten bioactive compounds were selected on the basis of their biological activities and further investigated using in silico molecular docking simulation to correlate with the identified pharmacological properties. Additionally, the ADME properties of the compounds were evaluated according to their drug-likeness profile. MECP had a maximum total phenol content of 209.85 ± 3.40 gallic acid equivalents/g extract and a total flavonoid content of 105.17 ± 3.45 quercetin equivalents/g extract, with an IC₅₀ value of 631.44 μg/mL. MECP (62.5-500 μg/mL) elicited 20.96-38.12% decreased protein denaturation compared to diclofenac sodium (65.40-83.50%), while a 35.72% (P < 0.001) clot lysis activity was observed for the 10 mg/mL concentration. MECP induced a dose-dependent reduction in locomotor activity, with a significant anxiolytic effect. In the analgesic test, MECP (200, 400 mg/kg) showed a 45.12% and 58.82% inhibition in analgesia, and the 400 mg/kg dose elicited a 27.5% inhibition in intestinal motility. These findings suggest that MECP might be effective in treating antioxidant, anti-inflammatory, and neuropharmacological defects, but this requires further study.

Keywords: Alternative medicine; Analgesic; Anti-inflammatory; Antioxidant; Anxiolytic; Bioinformatics; Cycas pectinata; Evidence-based medicine; Pharmaceutical science; Plant biology.

Figure 1

DPPH scavenging activity of methanol extract of C. pectinata leaves (MECP) extract compared to the standard ascorbic acid. Values are represented in Mean ± SEM (n = 3). ^$P < 0.001 statistically significant in comparison to Ascorbic acid followed by unpaired t-test (GraphPad Prism 7). MECP: Methanol extract of Cycas pectinata leaves.

Figure 2

Percentage inhibition of protein denaturation of methanol extract of C. pectinata leaves (MECP) extract compared to the standard diclofenac sodium. Values are represented in Mean ± SEM (n = 3). ^$P < 0.001 statistically significant in comparison to diclofenac sodium followed by unpaired t-test (GraphPad Prism 7). MECP: Methanol extract of Cycas pectinata leaves.

Figure 3

(A) The clot lysis activity of methanol extract of C. pectinata leaves (MECP) extract, streptokinase (SK) and normal saline (control). (B) The healthy volunteer status presented. Values are represented in Mean ± SEM. ^$P < 0.001 statistically significant in comparison to normal saline (control) followed by unpaired t-test (GraphPad Prism 7). MECP: Methanol extract of Cycas pectinata leaves.

Figure 4

(A) Locomotor activity of methanol extract of C. pectinata leaves and diazepam on open field test. (B) Locomotor activity of methanol extract of C. pectinata leaves and diazepam on hole-cross test. Values are represented in Mean ± SEM (n = 5). ^∗P < 0.05, ^#P < 0.01 and ^$P < 0.001 statistically significant in comparison to control followed by unpaired t-test (GraphPad Prism 7). MECP: Methanol extract of Cycas pectinata leaves.

Figure 5

(A) Anxiolytic activity of methanol extract of C. pectinata leaves and diazepam on % of time spend in open arm in elevated plus maze test. (B) Effect of methanol extract of C. pectinata leaves and diazepam on % of entry in open arm in elevated plus maze test. (C) Anxiolytic activity of methanol extract of C. pectinata leaves and diazepam on number of head-dipping in hole-board test. (D) Effect of methanol extract of C. pectinata leaves and diazepam on latency of first head-dipping in hole-board test. (E) Anxiolytic activity of methanol extract of C. pectinata leaves and diazepam on time spent in light compartment in light-dark test. (F) Effect of methanol extract of C. pectinata leaves and diazepam on number of transition in light-dark test. Values are represented in Mean ± SEM (n = 5). ^#P < 0.01 and ^$P < 0.001 statistically significant in comparison to control followed by unpaired t-test (GraphPad Prism 7). MECP: Methanol extract of Cycas pectinata leaves.

Figure 6

Analgesic activity of methanol extract of C. pectinata leaves and diclofenac sodium (DS) on acetic acid-induced writhing test with percentage of inhibition of analgesia. Values are represented in Mean ± SEM (n = 5). ^#P < 0.01 and ^$P < 0.001 statistically significant in comparison to control followed by unpaired t-test (GraphPad Prism 7). MECP: Methanol extract of Cycas pectinata leaves; DS: Diclofenac sodium.

Figure 7

2D representation of best docking score compounds: (A) 3,4-Dihydroxymandelic acid- 4WCU, (B) 3-Methylsalicylic acid-1A5H, (C) 13-Docosenamide, (Z)--5I6X, (D) 3,4-Dihydroxymandelic acid--4UUJ, (E) 3-Methylsalicylic acid--2OYE, (F) 2,5-Dihydroxybenzoic acid--6COX and (G)13-Docosenamide, (Z)--4U14.