Gelidiella acerosa protects against Aβ 25–35-induced toxicity and memory impairment in Swiss Albino mice: an in vivo report

Figures & data

Table 1. Details regarding dosage and treatment of Aβ 25–35 peptide and G. acerosa benzene extract in Swiss Albino mice.

Groups	Treatment	Dose
I	Vehicle control	1% Carboxymethyl cellulose (CMC) + PBS
II	Aβ 25–35 treatment	10 μM Aβ 25–35 in PBS
III	Aβ 25–35 + Extract treatment (Low dose)	10 μM Aβ 25–35 in PBS +200 mg/kg BW of G. acerosa benzene extract in CMC
IV	Aβ 25–35 + Extract treatment (High dose)	10 μM Aβ 25-35 in PBS +400 mg/kg BW of G. acerosa benzene extract in CMC
V	Extract alone	400 mg/kg BW of G. acerosa benzene extract in CMC
VI	Aβ 25–35 + Donepezil	10 μM Aβ 25–35 in PBS +1 mg/kg BW of donepezil in CMC
VII	Donepezil alone	1 mg/kg BW of donepezil in CMC

Figure 1. Effect of G. acerosa benzene extract on Aβ 25–35-induced short-term memory by Y-maze test. The results are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (Carboxy methyl cellulose (CMC) treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 2. Protective effect of G. acerosa benzene extract on Aβ 25–35-induced memory impairment by water-maze test. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 3. Effect of G. acerosa benzene extract on Aβ 25–35-induced memory impairment in mice by step-down inhibitory avoidance test. STM: short term memory; LTM: long term memory. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 4. Gelidiella acerosa benzene extract restores the alteration in the level of catalase (A) and SOD (B) activity in mice treated with Aβ 25–35. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kgbw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 5. Effect of G. acerosa benzene extract on Aβ 25–35 induced alteration in the level of glutathione-S-transferase (GST) and glutathione peroxidase (GPx) (A), glutathione reductase (GR) and glutathione (B). *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 6. Effect of G. acerosa benzene extract on Aβ 25–35-induced lipid peroxidation in brain. The values are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 7. Effect of G. acerosa benzene extract on Aβ 25–35-induced protein carbonyl content production in mice brain. The values are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 8. Inhibitory effect of G. acerosa benzene extract on Aβ 25–35 induced increase in acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) activities in mice brain tissue homogenate. The values are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 9. Inhibitory effect of G. acerosa benzene extract on Aβ 25–35 induced increase in β-secretase activity in mice brain tissue homogenate. The values are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 10. Inhibitory effect of G. acerosa benzene extract on Aβ 25–35 induced increase in monoamine oxidase activity in mice brain tissue homogenate. The values are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 11. Inhibitory effect of G. acerosa benzene extract on Aβ 25–35 induced increase in caspase-3 activity in mice brain tissue homogenate. The values are expressed as Mean ± SD. *p < 0.05 [Comparisons were made between groups II (Aβ 25–35 peptide treated) Vs I (CMC treated) & III (Aβ 25–35 peptide +200 mg/kg of extract in CMC), IV (Aβ 25–35 peptide +400 mg/kg of extract in CMC), V (400 mg/kg bw of extract), VI (Aβ 25–35 peptide + donepezil), VII (1 mg/kg bw of donepezil) Vs II (Aβ 25–35 peptide treated)].

Figure 12. Representative western blot images showing the effect of G. acerosa benzene extract on the expression levels of Bax and BCl-2 in mice brain tissue homogenates (A). Quantified results of the expression pattern of Bax (B) and BCl-2 (C).

Table 2. Organ weight of mice treated with G. acerosa benzene extract (2000 mg/kg of BW).

Organ	Control^a	G. acerosa (2000 mg/Kg BW of mice)^a
Kidneys^b	1.433 ± 0.3512	1.233 ± 0.152
Liver^b	5.167 ± 0.3512	5.4 ± 0.26
Heart^b	0.4333 ± 0.05774	0.433 ± 0.115
Lungs^b	0.5667 ± 0.05774	0.623 ± 0.351
Spleen^b	0.5833 ± 0.03512	0.536 ± 0.0152
Pancreas^b	0.3667 ± 0.05774	0.233 ± 0.057
Brain^b	0.48 ± 0.02	0.4567 ± 0.047
Ovary^b	0.26 ± 0.03464	0.2867 ± 0.030

^a Results were expressed as Mean ± S.D.

^b g/100 g of BW.

Table 3. Haematological profile of mice treated with G. acerosa benzene extract (2000 mg/kg of BW).

Parameters	Control^a	Gelidiella acerosa (2000 mg/Kg BW of mice)^a
Total RBC count (1 × 10^6 μL)	7.557 ± 0.3636	7.6 ± 0.26
Total WBC count (1 × 10^3 μL)	5.793 ± 0.1607	5.2 ± 0.305
Platelet count (1 × 10^3 μL)	179 ± 6.245	174 ± 4
Packed cell volume (%)	41 ± 3	42.67 ± 3.055
HB (g/dL)	12.4 ± 0.8	11.37 ± 0.15
MCV (fl)	47.33 ± 5.686	43.67 ± 2.0
MCH (pg)	6.867 ± 0.2517	6.33 ± 0.25
Neutrophils (%)	69.67 ± 3.215	66 ± 2.64
Lymphocytes (%)	77.33 ± 5.686	76.67 ± 2.08
Eosinophils (%)	0.3333 ± 0.5774	0.666 ± 0.577
Monocytes (%)	1.667 ± 0.5774	1.33 ± 0.577

^a Results were expressed as Mean ± S.D.

Figure 13. Histopathological analysis of the effect of G. acerosa on mice treated with 2000 mg/kg bw. A, B, C & D – Control group; E, F, G, H – G. acerosa-treated group. A&E-

Liver section showing normal hepatocyte architecture without any necrosis. B&F-

Heart muscles showing intact fibre alignment with normal tissue integrity. C&G-

Kidney sections showing normal tubules and interstitium. D&H-

Brain section with normal marginal alignment and interneuronal space.

Table 4. Biochemical profile of mice treated with G. acerosa benzene extract (2000 mg/kg of BW).

Parameters	Control^a	Gelidiella acerosa (2000 mg/Kg BW of mice)^a
Blood sugar (mg/dL)	78.33 ± 4.163	71 ± 2
BUN (mg/dL)	24.33 ± 3.055	24.33 ± 2.08
Serum creatinine (mg/dL)	0.8467 ± 0.0602	0.76 ± 0.12
Alkaline phosphatase (U)	100.7 ± 3.512	99.67 ± 3.055
Serum total protein (g/dL)	5.4 ± 0.9644	5.16 ± 0.230
Serum albumin (g/dL)	2.967 ± 0.2082	2.4 ± 0.2
SGOT (AST) (IU/mL)	54 ± 3.606	52.33 ± 3.215
SGPT (ALT) (IU/L)	60 ± 2.646	64 ± 1
Calcium (mmol/L)	6.933 ± 0.3786	7.433 ± 0.30
Sodium (mmol/L)	137.7 ± 4.509	154.3 ± 3.055*
Potassium (mmol/L)	5.267 ± 0.2082	5.4 ± 0.45
Bilirubin total (mg/dL)	0.7 ± 0.1732	0.733 ± 0.115
Serum total cholesterol (mg/dL)	85 ± 4.583	87.33 ± 1.52
Serum triglycerides level (mg/dL)	54.67 ± 3.512	53.83 ± 2.079
Serum HDL cholesterol (mg/dL)	66.67 ± 5.859	64 ± 1.732
Serum LDL cholesterol (mg/dL)	12.33 ± 3.215	11 ± 1
Serum VLDL cholesterol (mg/dL)	10.2 ± 1.908	10.3 ± 0.984

^a Results were expressed as Mean ± S.D. *p < 0.05.

Table 5. Organ weight of mice treated with G. acerosa benzene extracts (200 and 400 mg/kg of BW).

Organ	Control^a	Gelidiella acerosa 200 mg/kg of BW^a	Gelidiella acerosa 400 mg/kg of BW^a
Kidneys^b	1.383 ± 0.132	1.203 ± 0.07	1.533 ± 0.103
Liver^b	5.6 ± 0.229	5.6 ± 0.27	5.65 ± 0.372
Heart^b	0.483 ± 0.147	0.6 ± 0.109	0.633 ± 0.0816
Lungs^b	0.6533 ± 0.019	0.665 ± 0.025	0.68 ± 0.0275
Spleen^b	0.553 ± 0.041	0.5683 ± 0.021	0.5733 ± 0.02251
Pancreas^b	0.2 ± 0.0632	0.166 ± 0.081	0.233 ± 0.051
Brain^b	0.465 ± 0.019	0.5567 ± 0.022	0.5667 ± 0.0265
Ovary^b	0.37 ± 0.02	0.34 ± 0.045	0.37 ± 0.0219

^a Results were expressed as Mean ± S.D.

^b g/100 g of BW.

Table 6. Haematological profile of mice treated with G. acerosa benzene extract (200 and 400 mg/kg of BW).

Parameters	Control^a	200 mg/kg of BW^a	400 mg/kg of BW^a
Total RBC count (1 × 10^6 μL)	7.983 ± 0.42	8.45 ± 0.36	8.38 ± 0.41
Total WBC count (1 × 10^3 μL)	5.033 ± 0.47	5.81 ± 0.41	6.31 ± 0.56
Platelet count (1 × 10^3 μL)	178.8 ± 3.65	177 ± 5.83	177.2 ± 5.94
Packed cell volume (%)	44.17 ± 3.43	46 ± 1.89	46.5 ± 2.88
HB (g/dL)	11.55 ± 0.187	11.55 ± 0.30	11.85 ± 0.36
MCV (fl)	46.67 ± 2.5	48.5 ± 2.4	46.5 ± 2.0
MCH (pg)	6.48 ± 0.27	6.76 ± 0.25	6.833 ± 0.12
Neutrophils (%)	67.5 ± 2.07	69 ± 1.897	70.17 ± 5.41
Lymphocytes (%)	75.83 ± 2.85	75.17 ± 3.18	77 ± 2.75
Eosinophils (%)	0.333 ± 0.416	0.666 ± 0.526	0.67 ± 0.306
Monocytes (%)	1.167 ± 0.408	1.57 ± 0.547	1.333 ± 0.56
Basophils (%)	0.166 ± 0.408	0.333 ± 0.051	0.31 ± 0.16

^a Results were expressed as Mean ± S.D.

Table 7. Biochemical profile of mice treated with G. acerosa benzene extract (200 and 400 mg/kg of BW).

Parameters	Control^a	200 mg/kg of BW^a	400 mg/kg of BW^a
Blood sugar (mg/dL)	74.5 ± 5.5	82.83 ± 5.49	85.17 ± 2.78
BUN (mg/dL)	27.3 ± 2.42	26.5 ± 2.81	26.83 ± 2.63
Serum creatinine (mg/dL)	0.8833 ± 0.39	0.901 ± 0.070	0.931 ± 0.64
Alkaline phosphatase (U)	104 ± 4	102.3 ± 1.966	101.2 ± 3.54
Serum total protein (g/dL)	5.317 ± 0.174	6.117 ± 0.523	6.133 ± 0.54
Serum albumin (g/dL)	2.65 ± 0.18	3.05 ± 0.295	3.4117 ± 0.343
SGOT (AST) (IU/mL)	60.83 ± 4.16	62.67 ± 3.882	67.68 ± 3.50
SGPT (ALT) (IU/L)	65.33 ± 2.50	66 ± 3.28	66.33 ± 2.94
Calcium (mmol/L)	7.25 ± 0.48	7.56 ± 0.265	7.68 ± 0.14
Sodium (mmol/L)	152.7 ± 2.066	156.3 ± 2.33	163.5 ± 3.61*
Potassium (mmol/L)	5.667 ± 0.265	5.6 ± 3.033	5.917 ± 0.098
Bilirubin total (mg/dL)	0.8 ± 0.167	0.6167 ± 0.1169	0.8 ± 0.154
Serum total cholesterol (mg/dL)	85.17 ± 2.137	85 ± 4.817	83.33 ± 2.25
Serum triglycerides level (mg/dL)	55 ± 2.28	55.17 ± 3.25	54.5 ± 3.98
Serum HDL cholesterol (mg/dL)	65.5 ± 2.58	65.67 ± 3.14	67.17 ± 2.99
Serum LDL cholesterol (mg/dL)	10.33 ± 1.21	13.33 ± 2.33	12 ± 1.89
Serum VLDL cholesterol (mg/dL)	10.67 ± 1.211	11 ± 2	10.83 ± 1.83

^a Results were expressed as Mean ± S.D. *p < 0.05.

Figure 14. Histopathological analysis of the effect of G. acerosa on mice treated with 200 and 400 mg/kg bw. A, B, C, & D – Control group; E,F,G&H – 200 mg/kg bw G. acerosa-treated group, IJKL - 400 mg/kg bw G. acerosa treated group. A,E & I

- Liver section showing normal hepatocellular arrangement with intact membrane. B,F & J

- Heart section showing normal fibre integrity and cell alignment. C, G & K

- Kidney section showing normal interstitium. D, H & L

– Brain section shows normal interneuronal space with normal cellular integrity.