## 4 Discussion  
GE, belonging to the Orchidaceae family [50], has been shown to be effective in the cardiovascular system [51], convulsions [52], skin aging [53], and cancer cell invasion [54]. Above all, the beneficial neurological effects of GE extract and its constituents, especially gastrodin, have been revealed in A\(\beta\)-induced neuronal apoptosis [23], mice with pharmacologically induced memory impairments [55], and in a mouse model of AD [56]. GU belongs to the Leguminosae family [57] and has been determined to have memory enhancing [58], neuroprotective [48], and cerebral infarction modulative [59] effects. Our previous study demonstrated that GGW73 in mixtures of various proportion ratios (GER: GUR = 5:5, 6:4, 7:3, 8:2, and 9:1) had neuroprotective effects on A\(\beta\) stimulation in vitro [60]. Based on our previous results, we chose to use GGW73 for scopolamine-induced cognitive disorders in vivo. The Y-maze test is an experimental tool for measuring cognitive function in scopolamine-treated mice in vivo [61]. The Y-maze test, which uses the propensity of rodents to enter an unfamiliar place, is a widely used method to estimate the extent of memory impairment in cognitive disorders, such as AD and memory deficiency [62]. The total arm entries were significantly decreased by GGW73 100 mg/kg oral administration compared with scopolamine 1 mg/kg peritoneal injection (non-treatment group:12.60 \(\pm\) 1.21, scopolamine group: 26.00 \(\pm\) 0.84, GGW73 100 mg/kg group: 13.00 \(\pm\) 4.56, donepezil group: 19.00 \(\pm\) 1.53). Additionally, we observed decreased alternation triplets after scopolamine treatment in the Y-maze test and elevated alternation triplets by GEW 100 mg/kg and GGW73 100 mg/kg administration (non-treatment group: 65.22 \(\pm\) 6.22%, scopolamine 1 mg/kg group: 35.72 \(\pm\) 9.06%, GER 100 mg/kg group: 61.21 \(\pm\) 3.11%, GGW73 100 mg/kg group: 69.84 \(\pm\) 9.65%, donepezil group: 45.39 \(\pm\) 6.19%). These results indicated that oral administration of GGW73 (100 mg/kg) attenuated scopolamine-induced cognitive disorder. The water maze test has been representatively performed to evaluate memory and cognition in behavioral neuroscience [63]. The latency to reach the platform was reduced by GUR 100 mg/kg administration compared to scopolamine injection. Oral administration of GGW73 tended to shorten the latency, but no significant difference was observed (non-treatment group: 45.42 \(\pm\) 9.99 s, scopolamine group: 80.16 \(\pm\) 9.85 s, GUR 100 mg/kg group: 40.55 \(\pm\) 6.43 s, GGW73 100 mg/kg group: 64.79 \(\pm\) 14.49 s, donepezil group: 25.28 \(\pm\) 8.363 s) (Figure 3A). The time spent swimming in the maze and platform ratios were also significantly regulated by GUR 100 mg/kg; however, oral administration of GGW73 100 mg/kg also showed a slight regulatory effect (Figure 3B,C). The mice learned to recognize aversive stimuli, thereby enhancing cognitive function in the passive avoidance test [64]. Therefore, the passive avoidance test is the mostly used in vivo experiment for evaluating cognitive function disorder, memory deficient [65]. The scopolamine 1 mg/kg intraperitoneal injection decreased the latency to target (non-treatment group: 180.0 \(\pm\) 0.0 s, scopolamine group: 71.4 \(\pm\) 37.6 s); however, GGW73 administration significantly recovered the latency to target (Figure 4). The results showed that administration of GGW73 has the potential to remedy scopolamine-induced cognitive impairment. The accumulation of A\(\beta\) peptides is related to cognitive function. In cognitive disorders, the expression level of A\(\beta\) is elevated by a decrease in amyloid precursor protein (APP) [66]. Scopolamine contributes to increased A\(\beta\) release, which is influenced by APP [67]. The level of A\(\beta\) peptide was elevated by scopolamine injection (non-treatment group:24.06 \(\pm\) 1.30 pg/mL, scopolamine group:38.14 \(\pm\) 5.23 pg/mL).