Attenuation of neurological deficit by a novel ethanolamine derivative in rats after brain trauma

Figures & data

Figure 1 bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate (FDES) structure.

Table 1 Experimental groups

	Experiment 1	Experiment 2
Experimental groups	Intact TBI Choline alfoscerate, 100 mg/kg Citicoline, 500 mg/kg FDES, 75 mg/kg FDES, 10 mg/kg	Intact TBI FDES, 10 mg/kg FDES (10 mg/kg) + scopolamine (6 mg/kg) Scopolamine, 6 mg/kg
Animals	Wistar male white outbred rats (N=10 in each group)

Abbreviations: TBI, traumatic brain injury; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Table 2 The schedule of the experiments

	Day 1	Day 2	Day 3	Day 4	Day 5	Day 6	Day 7
Morning: model TBI ↓ 1 hour after TBI: DA	12:00 pm LPT ↓ DA	12:00 pm DA	12:00 pm OFT ↓ LPT ↓ DA	12:00 pm DA	12:00 pm DA	12:00 pm DA	12:00 pm EPMT ↓ CT ↓ BWT

Abbreviations: TBI, traumatic brain injury; LPT, limb placing test; OFT, open field test; EPMT, elevated plus maze test; CT, cylinder test; BWT, beam walking test; DA, drug administration.

Table 3 The influence of the drugs used in the experiment on the function of the limbs after TBI in the “Limb placing” test (Experiment 1)

Group	Day 1	Day 3	Day 7
Intact	14 (14; 14)	14 (14; 14)	14 (14; 14)
TBI	0 (0; 1.5)	5 (4; 7)	8 (6; 8)
Choline alfoscerate, 100 mg/kg	2 (0.5; 3)	6 (5; 8)	9 (8; 12)*
Citicoline, 500 mg/kg	1 (0.75; 2.75)	8 (7; 10)*	11.5 (10.75; 12)***
FDES, 75 mg/kg	0 (0; 2.5)	10 (4.75; 10.25)*	9.5 (7.75; 11)
FDES, 10 mg/kg	2 (0; 4)	8 (7; 9)*	10 (8.5; 11.5)**

Notes: P<0.001, significant differences in comparison with healthy rats; *P<0.05, **P<0.01, *** P<0.001, significant differences in comparison with TBI group. FDES at the dose of 10 mg/kg and reference agents improved the responses of fore- and hind limbs for tactile and proprioceptive stimulation in rats on day 7 post-trauma in comparison with untreated animals.

Abbreviations: FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate; TBI, traumatic brain injury.

Table 4 OMA and EA of rats in the “Open field” test (Experiment 1)

Group	OMA	EA
Intact	24.4±3.8	7.3±1.8
TBI	2.4±1.5*	0.4±0.4*
Choline alfoscerate, 100 mg/kg	5.1±2.3	0.7±0.5
Citicoline, 500 mg/kg	10.1±2.5	1.1±0.4
FDES, 75 mg/kg	13.8±5.6	1.1±0.5
FDES, 10 mg/kg	23.9±5.4**	1.8±0.4

Notes: *P<0.01, significant differences in comparison with healthy rats; **P<0.01, significant differences in comparison with TBI group. TBI induces in rats a pronounced decline of OMA and EA by day 3 post-trauma. In the present study FDES at the dose of 10 mg/kg was only one agent that increased OMA in traumatized animals. None of the drugs increased EA in rats after brain trauma.

Abbreviations: OMA, overall movement activity; EA, exploration activity; TBI, traumatic brain injury; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Table 5 Time spent in open and closed arms and OMA of animals in the “Elevated plus maze” on day 7 post-injury (Experiment 1)

Group	Time in OA, seconds	Time in CA, seconds	OMA
Intact	22.0±7.5	151.9±9.9	12.1±2.1
TBI	6.0±4.2	171.9±4.8	14.1±3.1
Choline alfoscerate, 100 mg/kg	4.0±4.0	171.6±4.5	2.5±0.8*
Citicoline, 500 mg/kg	3.1±3.1	172.1±5.1	3.4±0.7*
FDES, 75 mg/kg	12.2±6.9	164.3±8.9	5.7±1.4
FDES, 10 mg/kg	19.8±12.9	153.0±17.3	7.1±2.4

Notes: *P<0.01, significant differences in comparison with the TBI group. There were no differences in EPM behavior between groups of healthy rats and TBI on day 7 post-trauma. Course administration of choline alfoscerate or citicoline led to decrease of OMA in traumatized rats. FDES at both doses did not cause this effect.

Abbreviations: OMA, overall movement activity; OA, open arm; CA, closed arm; EP)M, elevated plus maze; TBI, traumatic brain injury; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Table 6 The frequency of contralateral forelimb using by animals in the “Cylinder” test on day 7 post-injury (Experiment 1)

Group	CL  %
Intact	45.0±4.3
TBI	20.9±3.4*
Choline alfoscerate, 100 mg/kg	22.6±5.3
Citicoline, 500 mg/kg	24.9±6.2
FDES, 75 mg/kg	20.4±3.1
FDES, 10 mg/kg	36.3±5.0

Notes: *P<0.05, significant differences in comparison with healthy rats. None of the studied compounds increased the frequency of contralateral forelimb use. FDES at the dose of 10 mg/kg had a mild positive effect but it was statistically nonsignificant.

Abbreviations: FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate; TBI, traumatic brain injury.

Table 7 The severity of the sensorimotor deficit (SMD) of the contralateral forelimb and hindlimb of animals in the «Beam walking» test on day 7 post-injury (Experiment 1)

Group	SMD of forelimbs,%	SMD of hind limbs,%
Intact	1.5±0.5	2.7±0.7
TBI	29.2±3.4	25.5±4.9
Choline alfoscerate, 100 mg/kg	27.3±2.8	41.3+1.4**
Citicoline, 500 mg/kg	10.0±2.1***	26.8+3.9
FDES, 75 mg/kg	8.0±1.5***	15.0+1.4
FDES, 10 mg/kg	4.8±0.8***	12.6+1.8*

Notes: P<0.001, significant differences in comparison with healthy rats; *P<0.05, **P<0.01, ***P<0.001, significant differences in comparison with TBI group. TBI induces in rats fore- and hind limb sensorimotor deficit which could be detected in the “Beam walking” test on day 7 post-trauma. FDES at both doses and citicoline at the dose of 500 mg/kg improved the contralateral forelimb function in traumatized rats. FDES at the dose of 10 mg/kg was the only agent which led to the recovery of contralateral hind limb function.

Abbreviations: SMD, sensorimotor deficit; TBI, traumatic brain injury; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Figure 2 The influence of the drugs in the experiment on the function of the limbs after TBI in the “Limb placing” test (Experiment 2). Significant difference between groups: *P<0.05, ***P<0.001. Scopolamine abolished the beneficial effects of FDES in traumatized rats by day 7 post--TBI.

Abbreviations: FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate; TBI, traumatic brain injury.

Figure 3 OMA and EA of rats in the “Open field” test (Experiment 2). Significant difference between groups: ***P<0.001. Scopolamine like an FDES at the dose of 10 mg/kg increased OMA in traumatized rats. Simultaneous administration of both agents abolished this positive effect.

Abbreviations: OMA, overall movement activity; EA, exploration activity; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate; TBI, traumatic brain injury.

Figure 4 Time spent in open and closed arms and OMA of animals in the “Elevated plus maze” on day 7 post-injury (Experiment 2). Significant difference between groups: *P<0.001. Neither Scopolamine, nor FDES decreased OMA in traumatized rats, however, with the simultaneous administration this effect was observed.

Abbreviations: OMA, overall movement activity; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Figure 5 The frequency of contralateral forelimb use by animals in the “Cylinder” test on day 7 post-injury (Experiment 2). Significant difference between groups: ***P<0.001.

Abbreviations: CL, contralateral limb; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Figure 6 The severity of sensorimotor deficit (SMD) of the contralateral forelimb and hindlimb of animals in the “Beam walking” test on day 7 post-injury (Experiment 2). Significant difference between groups: *P<0.05, **P<0.01, ***P<0.001. Scopolamine and FDES separately decreased the degree of sensorimotor deficit in rats after brain trauma, but simultaneous administration of both agents decreased the positive effects of these compounds in this test

Abbreviations: SMD, sensorimotor deficit; FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.

Figure 7 Profile of pharmacological activity of the tested drugs.

Abbreviation: FDES, bis-{2-[(2E)-4-hydroxy-4-oxobut-2-eneoyloxy]-N,N-diethylethanolamine} butanedionate.