## 4 Discussion  
In this study, commercially available enzymes such as bromelain, alcalase, and viscosyme were used to hydrolyze SCT. The hydrolysis pattern of HSCT was checked using SDS-PAGE, and it was confirmed that the proteins of SCT were effectively hydrolyzed by the enzyme treatment. Next, to confirm the effect of the developed HSCT on improvement of memory, we induced memory impairment in rats via intraperitoneal administration of scopolamine. One of the important causes of memory impairment in AD is decreased activity of the cholinergic system in the brain tissue [37; 38]. Scopolamine, which exerts anti-musccarinic effects by acting as a muscarinic ACh receptor antagonist, increases the activity of AChE, which degrades ACh at neuronal junctions and reduces memory in the short term [39]. Because of these properties, scopolamine is often used to induce memory impairment to study memory improvement in animals [40]. When comparing the results of the scopolamine-untreated control group and scopolamine-treated group, we also found that intraperitoneal administration of scopolamine effectively induced memory decline in rats. There were no significant differences in the intake of feed and body weight changes in rats due to intraperitoneal administration of scopolamine, and no significant differences were observed in organ weights among the scopolamine-treated groups. These results were similar to those reported by [PERSON] et al. [41], who induced memory impairment by administering scopolamine to rats. Rats can recognize their position in a confined space or maze, and with repeated training, they can find an escape route or new areas [42]. Using these characteristics, animal behavioral tests, such as the Barnes maze test, NORT, and Y-maze test, are commonly performed to investigate memory in rats [43]. The Barnes maze test assesses memory by measuring the distance and time a rat takes to navigate in a limited space to find an escape location [44]. The NORT is a method for assessing the memory that utilizes a rat's ability to navigate towards a new object [45], and the Y-maze test is a method for assessing the memory that utilizes a rat's ability to navigate towards a new, previously unvisited area [46]. In the current study, we investigated the effect of HSCT consumption on memory improvement by performing Barnes maze, NORT, and Y-maze tests in rats whose memory was impaired by scopolamine. The results of the Barnes maze test showed that on the 4 th day of the training period, the exploration distance and exploration time to find the escape route were significantly decreased in the HSCT and donepezil groups than in the scopolamine group (Figure 2). The donepezil group received an additional dose of donepezil as a positive control. Donepezil has been reported to be effective against severe AD by inhibiting AChE and was approved by the US Food and Drug Administration in  
Figure 7: Effect of HSCT on acetylcholinesterasecholinesterase activity of brains of scopolamine-induced memory impairment in the rats. Mean values were analyzed using Student’s \(t\)-test; \({}^{*}p<0.05\) compared with the scopolamine group. control, control diet; scopolamine, control diet + scopolamine (0.7 mg/kg, i. p.); HSCT, HSCT diet + scopolamine (0.7 mg/kg, i. p.); donepezil, control diet + scopolamine (0.7 mg/kg, i. p.) + donepezil (1.0 mg/kg, i. p.); i. p., intraperitoneal injection. 2010 [47]. In animal studies, donepezil has been used as a positive control to investigate its memory-enhancing effects, although its side effects may be dose dependent and are mostly transient and mild [48; 49]. In the NORT study, short-term memory was significantly improved in the HSCT group compared with the scopolamine group (Figure 3); however, there was no statistically significant difference in long-term memory between the HSCT and scopolamine groups (Figure 4).