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. 2022 Feb 21:2022:8305271.
doi: 10.1155/2022/8305271. eCollection 2022.

Stephania japonica Ameliorates Scopolamine-Induced Memory Impairment in Mice through Inhibition of Acetylcholinesterase and Oxidative Stress

Md Yusuf Al-Amin  1 , Amitav Lahiry  1 , Rafia Ferdous  1 , Md Kamrul Hasan  1 , Md Abdul Kader  1 , Ahm Khurshid Alam  1 , Zahangir Alam Saud  2 , Md Golam Sadik  1
Affiliations

Stephania japonica Ameliorates Scopolamine-Induced Memory Impairment in Mice through Inhibition of Acetylcholinesterase and Oxidative Stress

Md Yusuf Al-Amin et al. Adv Pharmacol Pharm Sci. .

Abstract

Alzheimer's disease (AD) is a progressive neurological disorder characterized by loss of memory and cognition. Stephania japonica is being used as traditional medicine in the treatment of different neurological problems. In this study, we evaluated the anticholinesterase and antioxidant activities of the crude methanol extract of S. japonica and its fractions in vitro and the neuroprotective effect of the most active fraction in the scopolamine-induced mouse model of memory impairment. Among the crude extract and its fractions, chloroform fraction exerted strong inhibition of acetylcholinesterase and butyrylcholinesterase enzymes with IC50 values of 40.06 and 18.78 µg/mL, respectively. Similarly, the chloroform fraction exhibited potent antioxidant activity and effectively inhibited the peroxidation of brain lipid in vitro. The phytochemical profile revealed the high content of polyphenolics and alkaloids in the chloroform fraction. Pearson's correlation studies showed a significant association of anticholinesterase and antioxidant activity with alkaloid and phenolic contents. Kinetic analysis showed that the chloroform fraction exhibited a noncompetitive type of inhibition. In experimental mice, the chloroform fraction restored the impaired learning and memory induced by scopolamine as evidenced by a significant decrease in latency time and increase of quadrant time in probe trial in Morris water maze task. The chloroform fraction also significantly reduced the activity of acetylcholinesterase and oxidative stress in mice. Our results suggest that the chloroform fraction of S. japonica may represent a potential candidate for the prevention and treatment of AD.

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Conflict of interest statement

The authors declare that they have no known conflicts of interest.

Figures

Figure 1
Figure 1
A general overview of the animal study timeline.
Figure 2
Figure 2
Anticholinesterase activity of the crude methanol extract of Sjaponica and its fractions. (a) AChE inhibitory activity. IC50 (µg/mL): DON, 5.21 ± 0.18; CME, 104.13 ± 1.86; CHF, 40.06 ± 0.79; EAF, 159.57 ± 2.57; AQF, 308.57 ± 2.27; PEF, 453.17 ± 4.28. (b) BuChE inhibitory activity. IC50 (µg/mL): GAL, 7.91 ± 0.35; CME, 50.44 ± 1.10; CHF, 18.78 ± 0.69; EAF, 35.30 ± 0.92; AQF, 44.73 ± 1.40; PEF, 100.08 ± 2.12. Results are expressed as mean ± SD (n = 3). Means with different letters (a–f) differ significantly (P < 0.05). CME, crude methanolic extract; CHF, chloroform fraction; EAF, ethyl acetate fraction; AQF, aqueous fraction; PEF, petroleum ether fraction; AChE, acetylcholinesterase; BuChE, butyrylcholinesterase; DON, donepezil; GAL, galantamine.
Figure 3
Figure 3
Lineweaver–Burk plot for inhibition of AChE (a) and BuChE (b) by different concentrations of CHF. Results represent the average values (n = 3).
Figure 4
Figure 4
Antioxidant activity of CME of S japonica and its fractions. (a) DPPH radical scavenging activity. IC50 (µg/mL): CAT, 3.03 ± 0.15; CME, 24.12 ± 1.00; CHF, 5.01 ± 0.16; EAF, 19.10 ± 0.73; AQF, 44.57 ± 1.90; PEF, 48.48 ± 3.47. (b) Hydroxyl radical scavenging activity. IC50 (µg/mL): CAT, 9.00 ± 0.34; CME, 97.24 ± 1.09; CHF, 17.12 ± 0.23; EAF, 56.67 ± 0.63; AQF, 86.08 ± 1.88; PEF, 162.00 ± 1.76. (c) Ferric reducing power. At 100 µg/mL concentration, the absorbances are as follows: CAT, 2.539 ± 0.045; CME, 1.471 ± 0.027; CHF, 2.489 ± 0.039; EAF, 1.548 ± 0.033; AQF, 0.627 ± 0.020; PEF, 0.609 ± 0.014. (d) Total antioxidant capacity. At 100 µg/mL concentration, the absorbances are as follows: CAT, 2.147 ± 0.042; CME, 0.948 ± 0.022; CHF, 1.852 ± 0.035; EAF, 0.862 ± 0.018; AQF, 0.253 ± 0.016; PEF, 0.473 ± 0.016. (e) Lipid peroxidation inhibition. IC50 (µg/mL): CAT, 18.23 ± 0.54; CME, 92.91 ± 2.35; CHF, 33.42 ± 0.45; EAF, 137.70 ± 1.31; AQF, 157.63 ± 3.15; PEF, 248.20 ± 2.50. Catechin (CAT) was used as a reference standard. Results are expressed as mean ± SD (n = 3). Means with different letters (a–f) differ significantly (P < 0.05). CAT, catechin; CME, crude methanolic extract; CHF, chloroform fraction; EAF, ethylacetate fraction; AQF, aqueous fraction; PEF, petroleum ether fraction.
Figure 5
Figure 5
Effect of CHF on scopolamine-induced learning and memory impairments in mice by Morris water maze (MWM) task. (a) Changes in the mean latency time during 4 consecutive days of training for mice of different groups in the MWM task. (b) Average time spent in the platform quadrant by mice of different groups in the MWM probe trial. Each group was significantly different (P < 0.05) from the disease control group (scopolamine-treated group). Values were expressed as MLT ± STD, n = 6 mice for each group.
Figure 6
Figure 6
Effect of CHF on brain AChE activity and oxidative stress biomarkers in the scopolamine-induced mouse model. (a) AChE activity, (b) rGSH level, (c) MDA level, (d) SOD activity, and (e) CAT activity in the brain of different groups of mice. Means with different letters (a-f) differ significantly (P < 0.05), where all the drug-treated groups were significantly different (P < 0.05) from the disease control group (scopolamine-treated group). Values were expressed as mean ± STD, n = 6 mice for each group. AChE, acetylcholinesterase; NV, normal vehicle group; SCO, scopolamine; DON, donepezil; CHF, chloroform fraction; rGSH, reduced glutathione; MDA, malondialdehyde; CAT, catalase.
Figure 7
Figure 7
Chromatographic and spectroscopic characterization of the CHF. (a) Thin layer chromatography (TLC) profile, (b) UV spectrum, and (c) IR spectrum of CHF.
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