Neuro-protective potential of a vesicular system of a standardized extract of a new chemotype of Withania somnifera Dunal (NMITLI118RT+) against cerebral stroke in rats^*

Figures & data

Figure 1. Chemical structure of Withanolide A.

Figure 2. Overlay of Withanolide A and NMITLI118RT+ fingerprint.

Table 1. Development of blank and drug-loaded formulations and their particle size measurements.

	Composition (% w/w)		Conditions
Label	S	C	rpm	Temperature (°C)	Size Z avr. (nm)	PDI
L1	80	20	120	50	156.70 ± 20.0	0.534 ± 0.06
L2	80	20	130	50	116.00 ± 13.0	0.581 ± 0.03
L3	55.55	44.44	130	60	330.60 ± 22.0	0.734 ± 0.17
L4	88.88	11.11	130	50	112.40 ± 18.0	0.738 ± 0.09
L5	88.88	11.11	150	50	133.96 ± 12.0	0.335 ± 0.06
L6	95.25	4.76	140	50	134.34 ± 09.0	0.264 ± 0.04
NMITLI118RT+LF	95.25	4.76	140	50	142.60 ± 09.0	0.268 ± 0.04

Values are expressed mean ± SD (n = 3).

Table

Control (Sham)	This group includes fresh male SD rats where all surgical procedure was carried out as in MCAO model except insertion of nylon filament
MCAO group	All surgical procedure was carried out, middle cerebral artery was blocked by insertion of filament, and only vehicle (gum accacia) was given by oral route
NMITLI118RT+ treated  (50 mg/kg p.o.)	(a) Treatment was given 1 h prior to the ischemic insult
	(b) Treatment was given after 6 h of I/R injury
NMITLI118RT+LF (liposomal formulation of NMITLI118RT+) treated (5 mg/kg p.o.)	(a) Treatment was given 1 h prior to the ischemic insult
	(b) Treatment was given after 6 h of I/R injury

Table 2. Physical stability of developed system as evident from particle size measurement.

Batches of	Particle size Z avr. (d. nm)		Zeta potential (mV)
NMITLI118RT+LF	Initial	After 3 weeks	Initial	After 3 weeks
1	141.25 ± 09.46	140.51 ± 07.48	−31.20 ± 1.00	−31.08 ± 1.15
2	139.48 ± 11.12	138.73 ± 12.47	−28.63 ± 1.25	−28.49 ± 1.08
3	142.73 ± 07.21	142.28 ± 09.26	−31.53 ± 2.36	−31.39 ± 2.12

Values are expressed mean ± SD (n = 3).

Figure 3. IR spectra of (a) NMITLI118RT+ and (b) NMITLI118RT+LF.

Figure 4. DTA and TG curves for (a) NMITLI118RT+; (b) lipid mixture and (c) NMITLI118RT+LF.

Figure 5. TEM micrograph for NMITLI118RT+LF.

Table 3. Evaluation of protection provided with NMITLI118RT+ and NMITLI118RT+LF in MCAO model.

Response	NMITLI118RT+	NMITLI118RT+LF
MDA (% decrease)
Pre-treatment	50.66	42.50
Post-treatment	24.83	44.83
GSH (%increase)
Pre-treatment	63.29	49.38
Post-treatment	64.55	66.89
Neurological deficit scores (% decrease)
Pre-treatment	64.93	69.98
Post-treatment	37.65	75.44
Cerebral infarcts (% decrease)
Pre-treatment	81.00	85.00
Post-treatment	44.52	81.41

Figure 6. (a) Effect of pre-treatment (1 h) and (b) post-treatment (6 h) of NMITLI118RT+ and NMITLI118RT+LF on MDA level following I/R injury.

Figure 7. (a) Effect of pre-treatment (1 h) and (b) post-treatment (6 h) of NMITLI118RT+ and NMITLI118RT+LF on GSH level following I/R injury.

Figure 8. (a) Effect of pre-treatment (1 h) and (b) post-treatment (6 h) of NMITLI118RT+ and NMITLI118RT+LF on neurological deficit following I/R injury.

Figure 9. (a) Effect of pre-treatment (1 h) and (b) post-treatment (6 h) of NMITLI118RT+ and NMITLI118RT+LF on cerebral infarct following I/R injury.

Figure 10. (a) Brain slices stained with TTC showing the effect of pre-treatment (1 h) and (b) post-treatment (6 h) of NMITLI118RT+ and NMITLI118RT+LF following I/R injury.